GET WELL SOON

ADJUNCTIVE TUMOR & CANCER SUPPORT * 90 caps / 400 mg. All Life Stages (* as indicated by a holistic veterinarian) The complexity of the immune system requires a multidimensional approach of nutritional factors that support normal immune function. The immune system is not intended to be suppressed or stimulated, however, in cases of immune dysfunction, requires systemic support to return to a state of balance. GET WELL SOON contains plant botanicals that provide nutritional and biochemical support for healthy immune response and systemic health and strength as well as to enhance the growth of healthy cells and tissues in the body’s defense mechanism; used holistically for all types of cancer and tumors, to protect healthy cellular response, to protect against invasion by potential allergens and pathogens, for most types of viral and bacterial diseases and diseases and for all types of inflammation and auto immune diseases. GET WELL SOON is comprised of Graviola, containing Annonaceous acetogenins which have remarkable cytotoxic, anti-cancer, and anti-tumoural activity due to their ability to slow and block the production of energy of abnormal and cancerous cells, to prevent the growth of blood vessels in or around tumors, to deplete the DNA building blocks necessary for new, abnormal cell division and to kill MDR (multi-drug resistant) cells that are resistant to chemotherapy. GET WELL SOON is used holistically as an immune booster, for cancer, cysts and tumors, including fibrous, fatty, sebaceous tumors including cutaneous mast cell tumors (mastocytomas, mast cell tumors, and sarcomas), lipomas, histiocytomas, adenomas hyperplasia and papillomas. Also used adjunctively for the following types of tumors: hind quarter tumors, deep tissue tumors, for lesions, polyps, warts, for basal and mast cell tumors, for bone tumors, for brain tumors, for heart tumors, for liver tumors, for kidney tumors, for bladder tumors, for mammary tumors, for skin tumors, for stomach tumors, for eye tumors, for ear tumors, for nose tumors, for mouth tumors and for leg tumors. GET WELL SOON is also used holistically as an adjunctive cytotoxic (kills cancer cells) therapy against cancer cells and complementary therapy in cancer protocols due to its active content of Annonaceous acetogenins and novel compounds which have demonstrated significant anti-tumorous (slows growth), and anti-cancerous activity (inhibits anaerobic cells

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while protecting healthy cells), broad-spectrum internal and external antimicrobial, anti-bacterial, anti-fungal and anti-viral properties against infection. 19BGET WELL SOON has demonstrated in vitro toxicity to ovarian, cervical, breast, colon, lung, pancreas, bladder, and skin cancer cells. Graviola’s bioactive compounds and phytochemicals have been shown to selectively attack and destroy only malignant cancer cells while demonstrating no toxicity to healthy cells because the plant compounds act as superb inhibitors of enzyme processes that are only found in the membranes of cancerous tumor cells, whereas chemotherapy indiscriminately destroys all actively reproducing cells. 20BOther therapeutic indications include use to slow tumor growth, for its cellular protective properties, to restrict growth in some types of tumors, and for its affinity to work effectively against multi drug resistant (MDR) tumors and cancer cells by blocking the transfer of ATP while supporting and building immunity. GET WELL SOON! is also used holistically as an antihypertensive to reduce blood pressure, to lower heart rate and dilate blood vessels, as a cardio protector and cardio tonic, for fibromyalgia, for neuralgia and to reduce nerve pain and response. 21BAlso used holistically for muscle and joint tenderness associated with all types of arthritis and rheumatism, for liver ailments, as a digestive, to expel worms and parasites, and for its positive effects on depression, stress, anxiety, nervous disorders, dyspepsia and ulcers and as, antispasmodic and anti-convulsant. 22BGET WELL SOON: PLANT IDENTIFICATION 23BFamily: Annonaceae 24BGenus: Annona 25BSpecies: muricata 26BSynonyms: Annona macrocarpa, A. bonplandiana, A. cearensis, Guanabanus muricatus 27BCommon names: Graviola, soursop, Brazilian paw paw, guanábana, guanábano, 28Bguanavana, guanaba, corossol épineux, huanaba, toge-banreisi, durian benggala, nangka 29Bblanda, cachiman épineux 30BPart Used: Leaves, fruit, fruit, seeds, bark, roots, stem 31BSome ways GET WELL SOON may be used to benefit animal wellness: 32BCancer is among the most dreaded diseases for pets and according to the Whole Dog Journal, more than 50% of dogs over the age of 10 are diagnosed with cancer every year, and the incidence of cancer in cats and younger animals is growing. The most common origins of cancer include viruses, toxins in the home and environment, conventional medical treatments that can disrupt the immune system and cause vaccinosis (diseases originating from medications and vaccination), genetics, aging, poor nutrition, unhealthy lifestyle and stress. 33BAs cells divide, mutations arise. In most pets, these abnormal mutated cells are killed by their immune systems. Cancers arise when the immune system fails to kill the 34Bmutated cells, which typically occurs at a higher frequency in older pets. On a cellular level, cancer spreads through a series of cellular defects and genetic mutations within the cells. 35BRecent research suggests that cancer is primarily a chronic inflammatory disease where inflammation not only kills healthy cells directly, but also deposits toxic inflammatory by-products and other “sludge” in the extracellular matrix that surrounds the cells. This toxic build-up reduces the flow of oxygen, nutrients, and wastes between cells and blood, and creating a fertile environment for abnormal cells that can thrive in such damaged environments. Preventing and resolving inflammation and clearing the matrix are primary goals of any program to prevent or treat cancer.

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Cancer systemically attacks the immune system and GET WELL SOON can be particularly critical in severely immuno-compromised cancer patients. Graviola contains a variety of plant actions including immunostimulating properties that also not only support but build the immune system while also limiting or killing the cells that encourage cancer to to thrive. The research of Graviola has identified more than 40 compounds with anti-cancerous properties, capable of killing cancer cells. Graviola’s ability to kill cancer cells has been well-documented but there are other important plant actions that are essential in a cancer treatment regimen, including its ability to block ATP production. For example, one compound is named “bullatacin” which has been reported to kill MDR (multi-resistant cells) on human cancer cells by inhibiting the production of an ATP (adenosine triphosphate), high-energy molecules found in the cells of all living things. Cancer cells exploit the process of ATP, robbing the energy from healthy cells, enabling cancer cells to mutate and replicate unchecked. Graviola’s acetogenins contain a plant mechanism that deprives cancer cells of their energy supply by selectively limiting access to the ATP process for cancer cells only, thereby supporting healthy cell replication. Holistic veterinarians recommend nutraceutical products in cancer fighting regimens that may or may not also include chemotherapy, radation or surgery. Nutraceutical products are viewed as both stand-alone regimens as well as adjunctive or complementary regimens to chemo-radio/surgical options. The natural products recommended in holistic cancer regimens contain immunostimulating herbs, glycoproteins, antioxidants, Omega-3 fatty acids, and proanthocyanidins. Immunostimulating Herbs: A healthy body naturally performs the complicated function of recognizing foreign agents or germs, neutralizing them and repeating the response later when needed. A body lacking immune support cannot perform these functions and is at greater exposure to disease. Autoimmune diseases such as cancer occur commonly in dogs and sometimes in cats and results when a pet’s body forms antibodies and attacks its own tissue. Products that support the immune system and also support anti-cancerous and antitumor activities are strongly recommended in pets suffering from autoimmune failures such as cancer, including I Feel Good which is also a powerful anti-inflammatory and immune building plant. Glycoproteins: Scientific evidence exists that recommends using a solid tumor scavenging product that contains glycoproteins such as Joint Ease Super Dog & Cat or I’m Allergic to Needles. The sugars of mannose and glucose are readily absorbed by the body and research indicates that these specific types of sugars could have important non-toxic consequences on solid tumors in the adjunctive treatment of cancer. Most types of glycoconjugate sugars have demonstrated an ability to inhibit cancer growth and the spread of tumor cells both in vitro (lab based) and in vivo (experiments in pets and people). The ability of glycoproteins to inhibit tumor growth may be related to their ability to alter the activities of the immune system. Glycoconjugate sugars stimulate white blood cells (macrophages) which secrete interferons. The interferon activates natural killer cells that help eliminate cancer cells. The glycoproteins may inhibit the spread of tumor cells by preventing them from adhering to each other as a result of the competitive inhibition of glycoconjugate receptor binding. Antioxidants: An antioxidant is a molecule capable of inhibiting the oxidation, which is a chemical reaction that produces free radicals. Free radicals cause damage or death to healthy cells. Antioxidants terminate and remove the mechanism for free radical activity, but also inhibit other oxidative reactions, preventing further cellular degeneration, particularly critical in cancer or immuno-compromised pets. A recommended antioxidant includes All Shins & Grins for its concentration of antioxidants, vitamins A, B, C, E and K, plus iron, phosphorus, selenium, manganese, zinc, proteins, leucine, serine, valine, flavonoids, necessary enzymes and much more. All Shins & Grins also contains plant chemicals to support healthy skin, coat, bone and connective tissue as well as boost the immune system and helps to prevent colds and flus.

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43BAnother highly recommended form of antioxidant for cancer patients are found in Hepa Protect (which contains burdock, dandelion root, hawthorn and bilberry.) Hepa Protect is useful for its antioxidant activities but also for its liver, kidney, pancreas and bladder protecting plant compounds, particularly in cases of chemo-radio therapies. I AM A Rock Star! and Super Model in a Bottle are recommended as adjunctive antioxidant supports. 44BOmega 3 Fatty Acid Sources: Natura Petz offers Gland Candy, Yelp for Kelp and Joint Ease, products which contain fatty acids and lignans, which are currently being studied for use in preventing cancer in people and animals. Many species of pets cannot convert ALA to other forms of more active non-inflammatory omega 3 fatty acids, and therefore require supplementation in their diets. In transplanted tumor models, omega-3 fatty acids reduced tumor development and inhibited tumor growth as well as spread of cancer (metastasis). 45BReduced radiation damages in the skin were also seen following supplementation with Omega-3 fatty acids. Use of Omega-3 fatty acids may also promote weight gain and may have anticancer effects due to its immunostimulating effects as well as improves the metabolic status and clinical outcomes in cancer patients. Omega-3 fatty acids also reduce recovery time after surgery and complications in pets and people with gastrointestinal cancers. Also, cats cannot convert Flax Seed Oil for omega 3 usages, so fatty acids are recommended in all cases. 46BIn animal models, omega-3 fatty acids also inhibits the formation of tumors and metastasis as well as showed anticachetic (anti-wasting) or starvation effects that are commonly experienced with cancer. 47BProanthocyanidins: Naturally occurring polyphenolic compounds found in plants such as All Shins & Grins, Hepa Protect and I Want Liquid Immunity are also recommended holistically. Proanthyocyanidins are used for their antioxidant effects against lipid (fat) perodixidation and may potentiate the immune system (via enhancement of T-lymphocyte activity and modulation of the neutrophil and macrophages responses and are often recommended in the treatment of pets with cancer but also enhance immunity and strengthen connective tissue. Natura Petz offers All Shins & Grins and I Want Liquid Immunity which contains grape seed extract plus the cancer fighting Noni plant. 48BAnother recommendation is Hepa Protect, which includes the plants burdock, dandelion root, hawthorn and bilberry, which protect the liver, kidneys, pancreas and bladder, particularly useful in cases of chemotherapy and radiation. 49BConventional cancer treatments like radiation, surgery, and chemotherapy may destroy the cancer yet create other problems at the same time. And even the most cutting-edge therapies may only prolong a pet’s life without truly curing the cancer. Quality of life issues also impact the choice of treatments once cancer has invaded. 50BIn addition to supplementation, both prevention and treatment of cancer involve the same components:

1. 51BAny cancer prevention or treatment program begins with diet. A balanced, home-prepared diet of fresh, preferably organic, whole foods is necessary. When the body is supported with the building blocks needed to maintain healthy cells and repair damaged ones, healing from within can begin. Many types of cancer cells, particularly lymphoma, utilize glucose from carbohydrates as fuel. When you limit the fuel, the cancer’s growth will also be limited. A low-carbohydrate diet can be very helpful in fighting cancer. Additionally, many cancer cells cannot utilize fat as an energy source; so more and better quality fats in the diet will help combat the weight loss that commonly occurs in cancer patients. Holistic veterinarians frequently recommend a diet that is low-carbohydrate, moderate protein, and moderate to high fat for cancer patients. The most harmful carbohydrates come from processed grains, fructose-containing fruits, and starchy vegetables such as white potatoes and peas. Dry kibble must contain starch due to processing requirements. In general, dry food should be avoided. Providing an optimal diet based on

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fresh, whole foods is highly improbable with dry kibble as most commercial pet foods (especially dry foods) are made with the leftovers and unwanted parts from livestock slaughter and processing, and loaded with additives and preservatives. Veggies with higher levels of fiber and plenty of antioxidants are better choices. For dogs, a diet of roughly 50% meat, and 40–50% non-starchy vegetables or whole grains is optimal. Fish oils should be added to provide additional fat; omega 3 fats are best and are abundant in fish oil. For cats, a diet of 80% meat (cats can tolerate fattier meats than dogs) and 20% non-starchy vegetables is recommended, again with fish oil added for additional fat.

For both dogs and cats, supplementation with vitamins, minerals, marine-source Omega-3 essential fatty acids, and antioxidants should also be part of the daily diet. Both humans and pets alike can likely no longer lead a healthy existence without supplementation.

2. Use only clean, purified water as this is essential to the body’s ability to resolve inflammation and clean up its waste products.

3. Limit vaccinations; the antibodies produced by vaccines cause inflammation and every booster perpetuates it.

4. Reduce indoor air pollution (perfumes, dryer sheets, toxic cleaning materials), yard chemicals, and other sources of toxic exposure that will help calm an overly reactive immune system and allow the body to cleanse and heal.

5. Minimize electromagnetic radiation which is a cause of low-grade, chronic inflammation.

6. Use a safe, non-toxic flea control program that is natural such as Brewer’s Yeast or Shoo! Tags. Most flea products contain pesticides that can contribute to the toxic overload of the body and inhibit natural cleansing processes.

7. Minimize stress and provide adequate exercise are tools that boost the immune system.

8. If chemo-radio treatment is pursued to deal with pets with cancer, I Feel Good is strongly recommended as an adjunctive therapy to reduce the side effects of chemo-radio therapy as well as stimulate the immune system during and after treatment.

If your pet is a candidate for surgery and you are utilizing cat’s claw, please discontinue use 2 weeks prior to surgery as cat’s claw has a blood thinning effect on some pets. There is a growing holistic movement against over vaccination of cats and dogs. For Dr. Hershman and most holistic veterinarians, routine vaccinations top the list of things to avoid. “Vaccines really disrupt the immune system,” she says, “especially combination vaccines that are given annually.” Like many holistic veterinarians, she recommends a single-dose parvovirus vaccination at age 10 to 12 weeks, followed by a single-dose distemper vaccination four weeks later and a rabies vaccination after age six months. “I check the effectiveness of these shots with blood titer tests,” she says. “If immunity is strong, there’s no need to revaccinate. If it’s weak, I repeat whatever the puppy needs for protection.” Label directions warn veterinarians not to vaccinate a sick animal; Dr. Hershman includes injured or stressed animals in that caution.

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65B“Vaccinating a dog who’s being spayed, neutered, or treated for an injury is totally irresponsible,” she says. “You want the animal to be healthy, with a strong vital force, not in a weakened, vulnerable state, when you introduce substances designed to challenge the immune system.” 66BResponding to decades of research by immunologists, veterinary textbooks and colleges no longer recommend annual vaccinations for dogs, but most veterinary clinics continue to prescribe them. “They routinely prescribe antibiotics, steroids, and other symptom-suppressing drugs, too,” says Dr. Hershman, “and those take a toll on the immune system. Whenever you can use nutrition, homeopathy, acupuncture, medicinal herbs, or other natural therapies instead of symptom-suppressing drugs, you strengthen the dog’s immunity. A strong immune system is the best defense against cancer.” 67BGood genes 68BAn important first step in selecting a puppy or adult dog is learning everything you can about the immediate family – parents, grandparents, siblings, aunts, uncles, cousins, etc. Some breeds are notoriously prone to cancer, and some lines within those breeds reinforce the trend. Look for good genes and good health when selecting puppies or adopting adult dogs. 69BOf course, rescued dogs seldom come with this documentation, and even the best-bred dog can develop cancer. But starting with good raw material can reduce the risk – and if you know that your dog may be prone to certain types of cancer, do what you can, starting today, to make that diagnosis less likely. 70BSpaying/neutering 71BThe statistics are convincing: female dogs have a significantly lower risk of developing mammary tumors if they are spayed before coming into season for the first or second time, and testicular cancer is obviously not a problem in neutered males. 72BBut while early spaying reduces the risk of mammary cancer, it quadruples the risk of developing cardiac hemangiosarcomas (vascular tumors) compared to intact females. In addition, a study of 3,218 dogs neutered before one year of age showed that both males and females had a significantly increased chance of developing osteosarcoma (bone tumors) compared to intact males and females. 73BUnderstanding your dog’s inherited risks can help you make informed decisions about whether and when to schedule surgery. 74BVaccinations 75BFor Dr. Hershman and most holistic veterinarians, routine vaccinations top the list of things to avoid. “Vaccines really disrupt the immune system,” she says, “especially combination vaccines that are given annually.” Like many holistic veterinarians, she recommends a single-dose parvovirus vaccination at age 10 to 12 weeks, followed by a single-dose distemper vaccination four weeks later and a rabies vaccination after age six months. 76B“I check the effectiveness of these shots with blood titer tests,” she says. “If immunity is strong, there’s no need to revaccinate. If it’s weak, I repeat whatever the puppy needs for protection.” 77BLabel directions warn veterinarians not to vaccinate a sick animal; Dr. Hershman includes injured or stressed animals in that caution. “Vaccinating a dog who’s being spayed, neutered, or treated for an injury is totally irresponsible,” she says. “You want the animal to be healthy, with a strong vital force, not in a weakened, vulnerable state, when you introduce substances designed to challenge the immune system.” 78BResponding to decades of research by immunologists, veterinary textbooks and colleges no longer recommend annual vaccinations for dogs, but most veterinary clinics continue to prescribe them. “They routinely prescribe antibiotics, steroids, and other symptom-suppressing drugs, too,” says Dr. Hershman, “and those take a toll on

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the immune system. Whenever you can use nutrition, homeopathy, acupuncture, medicinal herbs, or other natural therapies instead of symptom-suppressing drugs, you strengthen the dog’s immunity. A strong immune system is the best defense against cancer.” The right diet Advocates of home-prepared diets for dogs have long claimed that their animals are healthier than they would be on commercial pet food. Beth Taylor and Steve Brown, authors of See Spot Live Longer, agree. They blame dry and canned dog foods for a host of problems because they usually contain inferior-quality proteins, fats, and carbohydrates, a variety of toxins, highly processed grains, chemical preservatives, allergens, and other questionable ingredients. Many veterinarians blame grain-based pet foods for diabetes, digestive problems, and other canine disorders. After all, the canine digestive tract evolved on a diet of prey animals, consisting mostly of meat and bones, not wheat and corn. Every few years, aflatoxin, which grows on corn, rice, and other grains, contaminates pet foods and kills dogs (see “Yes, Dog Food Can Kill,” February 2006). In addition to causing liver damage, aflatoxin is a potent carcinogen, so even “safe” levels that don’t cause obvious disease outbreaks can contribute, over time, to cancer. Another carcinogen found in grain-based foods is acrylamide, once believed to exist only in industrial waste. However, acrylamide has recently been found almost everywhere in the human diet. This tasteless, invisible by-product is formed when high-carbohydrate foods are fried or baked at high temperatures. French fries and potato chips contain the highest concentrations, but acrylamide occurs in breads and breakfast cereals as well. The U.S. Environmental Protection Agency (EPA) considers acrylamide so dangerous that it set the “safe level” for human consumption at almost zero, with the maximum safe level in drinking water set at 0.5 parts per billion. A small serving of French fries contains over 400 parts per billion. No one has tested pet foods, but any processed foods that contain carbohydrates, especially those extruded at high temperature like grain-based kibble or canned under high heat and pressure, pose a risk. “Considering how ubiquitous these carcinogens are,” says San Francisco-area dog health researcher Mary Straus, “and considering that cancer cells thrive on carbohydrates, avoiding grains altogether may be one way to help lower the risk of cancer.” In addition to reducing levels of carbohydrates and carcinogens, feeding a home-prepared diet of pasture-fed, organically produced ingredients (see “Upgrading to Pasture-Fed,” July 2003) insures that your dog will not ingest pesticide and drug residues. Food prepared at home from conventionally farmed ingredients may not be free of pesticide residues, but it is unlikely to contain chemical preservatives, artificial colors or flavors, or the by-products of high-heat processing. In his book, Work Wonders: Feed Your Dog Raw Meaty Bones, Australian veterinarian Tom Lonsdale observes, “We need more information about the cancer epidemic in domestic dogs. However, basic nutritional and medical principles tell us that diet is the likely main factor. Without waiting for extra information, and because cancer often takes years to develop, it’s best to start puppies on a cancer-prevention diet early. From the whelping box to the grave, let ‘Prevention, not treatment’ be our motto.” Environmental factors Take two individuals from the same litter of puppies of a breed or family that has a high cancer risk. Place one with a family of heavy smokers who live next to a busy highway, use lawn chemicals, drink fluoridated tap water, and have high-current power lines in the backyard. Place the other pup on a pristine organic farm. Feed

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both the same diet and let several years go by. You don’t have to be an oncologist to know which dog is more likely to develop cancer. 90BSecond-hand smoke is a serious problem for pets, especially those who spend hours every day at the feet of their smoking companions. “I tell all my clients who smoke that they’re putting their dogs’ health at risk,” says Dr. Hershman. “I saw it happen in my own family, and it breaks my heart. Second-hand smoke is as dangerous to dogs as it is to infants.” 91BBusy highways, driveways, parking lots, and areas where trucks and cars idle are dangerous for dogs because of gas and diesel exhaust. A dog’s nose is much closer to the ground – and exhaust pipes – than the human nose, so dogs are more likely to inhale damaging particles. 92BLawn treatments and agricultural chemicals are known to cause cancer in animals (see “Canine Cancer Crisis,” November 2005). Dogs pick up pesticides, herbicides, and other chemicals through their feet and, when they sniff the ground, through their noses. Keep your dog off the grass in chemically treated neighborhoods, and explore organic alternatives for your own lawn and garden. 93BEven household chemicals pose a threat to our canine companions. According to the U.S. Consumer Product Safety Commission, more than 150 chemicals found in the average home are linked to birth defects, cancer, and psychological abnormalities. If labels carry a “keep away from children and pets” warning, or if product labels suggest they should be used only in well-ventilated areas, look for alternatives. 94BFluoride has gotten such good press over the decades that most Americans think it’s essential for healthy teeth. It’s even added to some canine tooth pastes. But in many countries, fluoride is considered a hazardous industrial waste, and its use in water supplies is prohibited. In September 2005, eleven unions representing more than 7,000 scientists and researchers at the EPA called for a national moratorium on the fluoridation of America’s drinking water, citing cancer risks. 95BA December 2005 analysis of more than 22 million tap water quality tests, most of which were required under the federal Safe Drinking Water Act, found that water suppliers across the U.S. detected 260 contaminants in public tap water. Of the 141 unregulated contaminants detected in water supplies between 1998 and 2003, 52 are linked to cancer, 41 to reproductive toxicity, 36 to developmental toxicity, and 16 to immune system damage. Water contaminated with 83 agricultural pollutants, including pesticides and fertilizer ingredients, flows through the taps of over 200 million Americans in 41 states. 96BInstalling a water filter or using uncontaminated, unfluoridated bottled water sounds like a very good idea! So does avoiding fluoridated toothpaste. 97BRegarding sources of electromagnetic radiation, a study published in 1995 in the American Journal of Epidemiology compared dogs treated at a veterinary teaching hospital for histologically confirmed lymphoma. Electric wire codes and magnetic fields were measured at the homes of 93 diagnosed cases and 137 controls, and a correlation was found between magnetic fields emitted by power lines and electrical appliances and the incidence of lymphoma. Dogs living in homes with very high current codes had the highest risk, while dogs living in homes with buried or underground power lines had a lower risk. 98BImmunologist and veterinarian Richard Pitcairn, DVM, PhD, author of Dr. Pitcairn’s Complete Guide to Natural Health for Dogs & Cats, considers all sources of radiation (including repeated diagnostic X-rays) dangerous because their effects are cumulative in the body. He recommends that dogs not be allowed to rest near a color TV set. Fortunately, the new flat-screen TVs and computer monitors emit much lower levels of electromagnetic radiation than older cathode ray tube models. In general, the fewer electrical appliances in close proximity to pets, the better.

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Topical pesticides Anyone who lives where fleas, ticks, or mosquitoes are a problem knows what a challenge they can be. Unfortunately, topical and systemic pest-control products contribute to a host of health problems, including increased cancer risks. A well-balanced raw diet can help a dog repel parasites, but sometimes the attack is overwhelming. “I definitely prefer natural alternatives to pesticide sprays or products like Frontline, which make the entire dog toxic to biting parasites,” says Dr. Hershman. “But alternatives don’t always work. One of my patients is a raw-fed Search and Rescue dog who often picked up more than 200 ticks on training weekends. He’s a German Shepherd Dog, so finding and removing them all was a time-consuming, stressful challenge. After his owner tried every natural repellent we could find, none of which solved the problem, he now applies K9 Advantix, a systemic pesticide that repels fleas, ticks, and mosquitoes, on a reduced dosage schedule only when needed. “When it comes to cancer prevention,” she says, “the less often you use conventional pesticides, the better. A good diet and natural repellents are always worth trying first.” Natural treatments include Baby Love Bits (Brewer’s Yeast) and Sentry Natural Defense. Natural products must be used with caution just as synthetics. Cancer preventives Several holistic cancer treatments, such as those described in “What Are the Alternatives?” (February 2006), can be used to help healthy dogs remain cancer-free. The thinking here is that cancer cells develop all the time, even in healthy bodies, but they don’t create problems until conditions encourage their growth. Preventive treatments disrupt cancer cells before they take up residence in vulnerable parts of the body. Henry Lai, PhD, the University of Washington researcher who first tested artemisinin (an extract of Artemesia annua, or annual wormwood) on dogs with cancer, takes artemisinin as a preventive himself and has tested it on laboratory animals. “It is hard to recommend a protocol for cancer prevention,” he says, “but, based on studies on rats, a good dose could probably be somewhere between 8 milligrams of artemisinin per kilogram of body weight per day at the high end and 10 mg/kg once per week at the low end. I take 100 mg per day for 10 days each month. Even though this approach hasn’t been tested yet on humans or canines, I think it makes sense.” Following Dr. Lai’s example, a dog weighing 60 to 75 pounds could take 50 mg artemisinin for 10 days each month, and the amount could be increased or decreased as needed for larger and smaller dogs. Dietary supplements and Medicinal Herbs Antioxidant supplements, which help protect the body from damage by free radicals, have many health benefits, including cancer protection. Best-selling antioxidant supplements include vitamins A, C, and E, beta carotene, lycopene, and the mineral selenium. Bear in mind that some alternative cancer treatments, such as artemisinin, are not compatible with antioxidants. Other emerging dietary supplements include herbs and homeopathic medicines, which have been used traditionally and holistically for generations outside of Western cultures. Many of these herbs have been studied extensively and are used widely as adjunctive and primary cancer treatments for all types of cancer and for tumors. Products include Get Well Soon (Graviola) and I Feel Good (Cat’s Claw Extract), amongst others, and have large bodies of research and trials to back up use. Food-source antioxidants, vitamins, and other nutrients derived from whole foods are recommended by many holistic veterinarians because they are recognized as food by the body and are more easily assimilated than synthetic vitamins grown in a laboratory. The words “whole food” or “food source” indicate natural rather than synthetic ingredients.

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112BThere are literally dozens upon dozens of experts that have their favorite herbs for cancer prevention, but a few herbal products are so effective that they are recommended by almost everyone. 113B“Many mushrooms have anti-tumor and immune-stimulating activity,” says Carol Falck, VMD, of Pompano Beach, Florida. “They have been used medicinally for thousands of years in China and Japan, and they work very well for dogs.” 114BI Feel Good (Cat’s Claw Extract), Get Well Soon (Graviola) and Kick Start My Heart (Green Tea) are recommended herbs because they enhance cellular immune function, increases natural killer cell activity, and may inhibit some cancer cell lines. 115BAnother favorite supplement for dogs at risk of cancer is curcumin, says Dr. Falck. “Curcumin is the yellow pigment in turmeric, the spice that gives curry its distinctive color. Both turmeric and curcumin have been shown to inhibit tumor growth. I also like astragalus, an herb with strong immune-stimulating properties. I use several Chinese herbal formulas containing astragalus, depending on the patient, including Astragalus for Animals by Buck Mountain Botanicals.” 116BSuper Model in a Bottle contains garlic, a well known and often used natural remedy for its cancer-inhibiting properties. Small amounts of fresh minced garlic or aged garlic extract can be added to any dog’s dinner. Garlic is an ingredient in Herbal Compounds tablets created by Juliette de Bairacli Levy, whose Natural Rearing philosophy pioneered home-prepared diets and alternative medicine for animals. Garlic should be used cautiously in animals. Do not exceed recommended dosages. 117B“This formula is very antiseptic,” says Natural Rearing advocate Marina Zacharias, who imports the product from England. “It definitely helps the immune system.” 118BClosely related to herbal medicine is aromatherapy. San Diego holistic veterinarian Stephen Blake recommends massaging the paw pads of at-risk dogs once or twice per day with a drop of blended frankincense, sandalwood, and Douglas fir essential oils. For best results, use organic or wild crafted oils from reputable distributors as described in “Essential Information” (January 2005). “These essential oils are great for detoxification and for supporting the immune system,” he says. 119BExercise 120BAlthough few of us appreciate the important role it plays, the lymph system is a key factor in cancer prevention. Lymph is a clear fluid, similar to blood but lacking red blood cells. It contains the immune system’s lymphocytes (T-cells and B-cells) and circulates through channels that carry waste to the lymph nodes, filtering bacteria and other toxins. 121BThe more lymph circulation is impaired, the less efficiently the body removes toxins and the more favorable conditions are for the growth of cancer. Lymph circulation improves with active exercise and deep, diaphragmatic breathing. Gentle to vigorous brushing that moves from the feet to the heart is a simple addition to daily grooming that also stimulates lymph circulation. 122B“Exercise is so important,” says Dr. Falck. “Exercise stimulates the immune system and releases endorphins, and an added benefit of consistent exercise is increasing gastrointestinal motility, which helps normalize stools and eliminate toxins from the body. It also facilitates weight management, which is important because obesity is a risk factor for some types of cancer.”

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GET WELL SOON: TEST DRIVE WHAT IT CAN DO FOR YOUR PET Main Actions: anticancerous, antitumorous, antimicrobial, antiparasitic, hypotensive (lowers blood pressure), dilates blood vessels, as a mild sedative Primary Holistic Uses: for all types of cancer, as a ctyotoxic therapy, for all types of tumors, for MDR cancer cell lines, as an anti-mutagenic (cellular protector), anti-carcinogen (prostate), anti-parasite, antispasmodic, sedative (very mild), reduces symptoms of asthma, reduces hypertension, reduces symptoms of diabetes, reduces symptoms of liver disease, as a broad-spectrum internal and external antimicrobial to treat bacterial and fungal infections, for internal parasites and worms, for high blood pressure, for depression, stress, and nervous disorders, to reduce spasms and stop convulsions, to reduce fever, to kill viruses, to expel worms, to stimulate digestion, as a hypotensive, anti-bacterial, for high blood pressure, to treat infections caused by bacteria and parasites including leishmaniasis, a disease caused by parasites transmitted, through the bite of sand fleas, herpes, coughs and cancer, for vomiting and to empty the bowels Properties/Actions Documented by Research: antibacterial, anticancerous, antitumorous, cytotoxic, anticonvulsant, antidepressant, antifungal, antimalarial, antimutagenic (cellular protector), antiparasitic, antispasmodic, antitumorous, cardio depressant, emetic (causes vomiting), hypotensive (lowers blood pressure), insecticidal, sedative, uterine stimulant, vasodilator, anthelmintic, antimicrobial, antineoplastic, antiviral, astringent, febrifuge, hypotensive, insecticide, nervine, pectoral, piscicide, sedative, stomachic, vasodilator, vermifuge, multidrug resistant cancer cells line Other Properties/Actions Documented by Holistic Use: cardio tonic (tones, balances, strengthens the heart), decongestant, digestive stimulant, febrifuge (reduces fever), nervine (balances/calms nerves), pediculicide (kills lice), vermifuge (expels worms) Contraindications: Graviola has demonstrated uterine stimulant activity and is contraindicated during pregnancy. Graviola has demonstrated hypotensive, vasodilator, and cardio depressant activities in animal studies and is contraindicated for pets with low blood pressure. Animals taking antihypertensive drugs should check with their doctors before taking graviola and monitor their blood pressure accordingly (as medications may need adjusting). Graviola has demonstrated significant in vitro antimicrobial properties. Chronic, long-term use of this plant may lead to die-off of friendly bacteria in the digestive tract due to its antimicrobial properties and supplementing the diet with probiotics and digestive enzymes is advisable if this plant is used for longer than 90 days. Graviola is likely contraindicated in combination with MAO inhibitors and some prescription antidepressants. Check with your Vet first if your pet is taking prescription antidepressants or MAO inhibitor drugs prior to taking graviola. Drug Interactions: None reported. Graviola may potentiate antihypertensive and cardiac depressant drugs. May potentiate antidepressant and MAO-inhibitor drugs drugs. May interfere with CoQ10 or other ATP-enhancers.

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157BCautions: 158BDo not use if your pet is pregnant or nursing. Large dosages can cause upset 159Bstomach, nausea and vomiting. Do not exceed suggested dosages. Use caution if your pet has been described MAOI inhibitors or antidepressants. Graviola might potentiate symptoms of Parkinson’s disease due to high concentrations of annonacin. Discuss this with your Vet prior adding Graviola to your pets diet. 160BSuggested Dosage: (please confirm dosage according to life stage of dog or cat) 161BDogs & cats up to 12 pounds: ½ capsule, 1 time daily. 162BDogs & cats 13-25 pounds: 1 capsule, 1 time daily. 163BDogs & cats 26+ pounds: 1 capsule, two times daily. 164BThe following products are recommended to avoid when taking Graviola as these products encourage ATP energy (what cells require for energy and production): hydrogen peroxide, high dosages of Vitamin C, Vitamin E, CO-Q10, thyroid support products, L-cysteine, N-acetylsteine, Gluathione, Essiac and Flor-essence, Ozone treatments (hyperbaric oxygen is acceptable), Flax seed oil, Grapefruit seed extract (high Vitamin C), Whey protein. 165BGET WELL SOON: ALL SORTS OF INTERESTING FACTS ABOUT HOW THIS PLANT MIGHT BENEFIT PET HEALTH, ITS BIOLOGICAL ACTIVITIES AND STUDIES CONDUCTED 166BGraviola has a lengthy and rich history of use (estimated at 4,000+ years of use) in herbal medicine systems as well as lengthy recorded indigenous use across many cultures. Different parts of the Graviola tree including bark, root, leaves, fruit and seeds have medicinal uses in the traditional treatment of diabetes, catarrhs, indigestion, intestinal parasites and even certain types of cancer. Graviola has become a highly recognizable in the adjunctive treatment of holistic cancer regimens and is often prescribed by holistic veterinarians for pets diagnosed with cancer as well as for all types of tumors, mirroring its usage in humans. 167BThe primary ways Graviola is recommended holistically is as an anti-carcinogen, anti-tumor and anti-parasite. Graviola has been prove to be 10,000 times more effective than Adriamicyn, which is commonly used in chemotherapy regimens. Adriamicyn kills both abnormal and healthy cells and severely reduces the immune system, causing nausea, hair loss and often, cachexia. Graviola, in contrast, acts selectively, killing only cancerous cells, without damaging the healthy cells and typically incurs little to no side effects. 168BResearch has been published regarding Graviola’s ability to destroy cancer cells in at least 20 laboratory tests and thus far, specific acetogenins in graviola and/or extracts of Graviola have been reported to be selectively toxic in vitro to these types of tumor cells: lung carcinoma cell lines; human breast solid tumor lines; prostate adenocarcinoma; pancreatic carcinoma cell lines; colon adenocarcinoma cell lines; liver cancer cell lines; human lymphoma cell lines; and multi-drug resistant human breast adenocarcinoma. 169BHolistic vets have many applications of use for Graviola apart from adjunctive cancer regimen use. Graviola is used holistically for catarrh (inflammation of mucous membranes), to kill parasites, for diarrhea, for all types of cysts and tumors, for diabetes, as a mild sedative, antispasmodic, antispasmodic, anti-dysentery, antipyretic, vulnerary, as a sedative, antispasmodic, as a nervine for heart conditions, for coughs, flu, asthma, hypertension, as a cardio tonic, for neuralgia, rheumatism, and arthritis pain, 170BResearch in pets has linked Graviola with measurable results in holistically and traditionally treating 12 different types from cancer, including colon, prostate, breast, lungs and pancreatic cancers. 171BThe therapuetic dosage of graviola leaf, (which offers just as high of an amount of acetogenins as the root and almost as much as the seed) is reported to be 2-3 grams taken 3 or 4 times daily. Graviola products (capsules and tinctures) are becoming more widely available in the U.S. market, and now offered under several different

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manufacturers’ labels in health food stores. As one of graviola’s mechanisms of action is to deplete ATP energy to cancer cells, combining it with other supplements and natural products which increase or enhance cellular ATP may reduce the effect of graviola. The main supplement which increases ATP is a common antioxidant called Coenzyme Q10 and for this reason, it should be avoided when taking graviola. Graviola pulp is consumed extensively and is featured in canine and feline pet products including holistic shampoos, conditioners, detanglers, creams and tinctures. Since the 1940’s, scientists have been studying graviola and many bioactive compounds and phytochemicals have been discovered. Graviola’s many uses in natural medicine have been validated by scientific research which have demonstrated that the bark as well as the leaves had antitumorous, antiparasitic, pesticidal, antiprotozoal, antifeedant, anthelmintic, antimicrobial, hypotenisve, antispasmodic, anticonvulsant, vasodilator, smooth muscle relaxant and cardio depressant activities in animals. 1, 2, 31 According to Memorial Sloan Kettering, Graviola extracts show antiviral, antiparasitic, antirheumatic, astringent, emetic, antileishmanial and cytotoxic, antinociceptive, anti-inflammatory, and antihyperglycemic properties. Graviola is also effective against multidrug resistant cancer cells line. Mode of action studies in three separate laboratories have recently determined that these acetogenins are superb inhibitors of enzyme processes that are only found in the membranes of cancerous tumor cells. Interestingly, it is known through research studies done by Dr. H. Clark (1) that there is a relation between cancer and parasites. Cancer patients are often, if not always, infected with parasites (human as well as feline and canine), which create toxins inside them. Holistic vets that recommend Graviola be used in conjunction with an anti-parasitic regimen, such as Shake Ur Groove Thing, as it has managed to stop the progression of certain cancers and tumors. An interesting link between cancer and parasites has been evidenced in dogs and cats. Graviola is also used by holistic vets for specific cancers, including bone cancer. Traditional veterinary treatment of bone cancer often involves amputation, according to Bone Cancer Dog. For instance, if the cancer is found in a leg bone and has not yet spread to other parts of a dog's body, amputation of the affected bone offers a better chance of survival. The alternative use of graviola may prevent the necessity of amputation. A study conducted by Dr. Jerry L. McLaughlin, et al. found that the use of graviola "reduced tumor markers, reduced tumor sizes, and increased longevities among 94 cancer patients while causing minimal side effects." In addition, according to the "Oncology Nursing Forum," graviola is is also powerful "against cancers that are resistant to multiple drugs," which is commonly happens as cancer patients are subjected to repeated chemotherapy treatments over time.” (Graviola for Dog Bone Cancer | eHow.com) From the time researchers first discovered an antitumorous effect in the bark of the pacific yew tree and a novel chemical called taxol was discovered in its bark, it took 30 years of research by numerous pharmaceutical companies, universities, and government agencies before the first FDA-approved Taxol drug was sold to a cancer patient (which was based on the natural taxol chemical they found in the tree bark). With graviola, it has taken researchers almost 10 years to successfully synthesize (chemically reproduce) the main antitumorous chemical, annonacin. These acetogenin chemicals have a unique waxy center and other unique molecular energy properties which thwarted earlier attempts, and at least one major pharmaceutical company gave up in the process (despite knowing how active the natural chemical was against tumors). Most of the research on graviola focuses on a novel set of chemicals called Annonaceous acetogenins. Graviola produces these natural compounds in its leaf and stem, bark, and fruit seeds. Three separate research groups have confirmed that these chemicals have significant antitumorous properties and selective toxicity against various types of cancer cells (without harming healthy cells), having published eight clinical studies on their

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findings. Many of the acetogenins have demonstrated selective toxicity to tumor cells at very low dosages—as little as 1 part per million. Four studies were published in 1998 which further specify the chemicals and acetogenins in graviola which are demonstrating the strongest anticancerous, antitumorous, and antiviral properties. 180BPurdue University, in West Lafayette, Indiana, has conducted a great deal of the research on the acetogenins, much of which has been funded by The National Cancer Institute and/or the National Institute of Health (NIH). Thus far, Purdue University and/or its staff have filed at least nine U.S. and/or international patents on their work around the antitumorous and insecticidal properties and uses of these acetogenins. 181BIn 1997, Purdue University published information with promising news that several of the Annonaceous acetogenins were “. . . not only are effective in killing tumors that have proven resistant to anti-cancer agents, but also seem to have a special affinity for such resistant cells." In the 1997 clinical study, novel alkaloids found in graviola fruit exhibited antidepressive effects in animals. The acetogenins plant chemicals were also documented with antitumorous, antiparasitic, insecticidal, and antimicrobial activities. Mode of action studies in three separate laboratories have recently determined that these acetogenins are superb inhibitors of enzyme processes that are only found in the membranes of cancerous tumor cells which is why they are toxic to cancer cells but have no toxicity to healthy cells. 182BIn several interviews after this information was publicized, the head pharmacologist in Purdue's research explained how this worked. As explained, cancer cells that survive chemotherapy can develop resistance to chemo as well as to other, even unrelated drugs. This phenomenon is called multi-drug resistance (MDR). One of the main ways that cancer cells develop resistance to chemotherapy drugs is by creating an intercellular pump which is capable of pushing anticancer agents out of the cell before they can kill it. On average, only about two percent of the cancer cells in any given person might develop this pump—but they are the two percent that can eventually grow and expand to create multi-drug-resistant tumors. 183BSome of the latest research on acetogenins reported that they were capable of shutting down these intercellular pumps, thereby killing multi-drug-resistant tumors. Purdue researchers reported that the acetogenins preferentially killed multi-drug-resistant cancer cells by blocking the transfer of ATP—the chief source of cellular energy—into them. A tumor cell needs energy to grow and reproduce, and a great deal more to run its pump and expel attacking agents. By inhibiting energy to the cell, it can no longer run its pump. 184BWhen acetogenins block ATP to the tumor cell over time, the cell no longer has enough energy to operate sustaining processes—and it dies. Normal cells seldom develop such a pump; therefore, they don't require large amounts of energy to run a pump and, generally, are not adversely affected by ATP inhibitors. Purdue researchers reported that 14 different acetogenins tested thus far demonstrate potent ATP-blocking properties (including several found only in graviola). They also reported that 13 of these 14 acetogenins tested were more potent against MDR breast cancer cells than all three of the standard drugs (Adriamycin, vincristine, and vinblastine) they used as controls. 185BFour studies were published in 1998 which further specify phytochemicals and acetogenins which are demonstrating the strongest anticancerous, antitumorous, and antiviral properties.22–25 Thus far, specific acetogenins in graviola have been reported to be selectively toxic to these types of tumor cells: lung carcinoma cell lines;14,16–19 human breast solid tumor lines;17 prostate adenocarcinoma;22 pancreatic carcinoma cell lines;14,22,25 colon adenocarcinoma cell lines;14,15,25 liver cancer cell lines;26,27,29 human lymphoma cell lines;28 and multi-drug resistant human breast adenocarcinoma.30 Annonaceous acetogenins are only found in the Annonaceae family (to which graviola belongs). 186BResearchers verified graviola leaf’s hypotensive properties in rats again in 1991. 3. Several studies over time have demonstrated that leaf, bark, root, stem and seed extracts of graviola are antibacterial in vitro against many

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pathogens. 4-6 and that the bark has antifungal properties. 6-7. Graviola seeds demonstrated active antiparasitic properties in a 1991 study 8 and a leaf extract showed to be active against malaria in two more studies in 1990 and 1993. 9, 10 Graviola leaves, root and seeds demonstrated insecticidal properties, with the seeds demonstrating strong insecticidal activity in an early 1940-42 study. 11 In 1997, a clinical study involving novel alkaloids found in graviola fruit exhibited anti depressive effects on animals. 12. Three separate research groups have isolated these acetogenin compounds in graviola which have demonstrated significant antitumorous and anticancerous properties, and selective toxicity against various types of cancer cells (without harming healthy cells) publishing eight clinical studies on their findings.14–21 Many of the acetogenins have demonstrated selective toxicity to tumor cells at very low dosages—as little as 1 part per million. In Japan, an interesting in vivo study was published (March 2002), where mice with lung cancers were inoculated. One third received nothing (the control group), one third received the chemotherapy drug adriamycin, and one third received the main graviola acetogenin, annonacin (at a dosage of 10 mg/kg). At the end of two weeks, five of the six in the untreated control group were still alive and lung tumor sizes were then measured. The adriamycin group showed a 54.6% reduction of tumor mass over the control group—but 50% of the animals had died from toxicity (three of six). The mice receiving annonacin were all still alive, and the tumors were inhibited by 57.9%—slightly better than Adriamycin—and without toxicity. This led the researchers to summarize; “This suggested that annonacin was less toxic in mice. On considering the antitumor activity and toxicity, annonacin might be used as a lead to develop a potential anticancer agent.” Researchers in Taiwan reported in 2003 that the main graviola acetogenin, annonacin, was highly toxic to ovarian, cervical, breast, bladder and skin cancer cell lines at very low dosages saying; “. . . annonacin is a promising anti-cancer agent and worthy of further animal studies and, we would hope, clinical trials.” A study at the Catholic University of South Korea revealed two chemicals extracted from Amazonian Graviola seeds showed comparable results to the chemotherapy drug Adriamycin when applied to malignant breast and colon cells in test tubes. 2 Another study, published in the Journal of Natural Products, showed that Graviola is not only comparable to Adriamycin – but dramatically outperforms it in laboratory tests. Results showed that it selectively killed colon cancer cells at “10,000 times the potency of Adriamycin.” 3 Perhaps the most significant result of the studies researched found that Graviola selectively seeks out and destroys cancer cells, leaving all healthy, normal cells untouched. Chemo radio therapy indiscriminately seeks and destroys all actively reproducing cells, even healthy, normal hair and stomach cells, causing such devastating side effects as hair loss and severe nausea. Natura Petz encourages you to consult with your vet before beginning any new therapy, especially it involves cancer. Several studies by different researchers demonstrated that the bark as well as the leaves had hypotensive, antispasmodic, anticonvulsant, vasodilator, smooth muscle relaxant, and cardio depressant activities in animals. Researchers verified graviola leaf's hypotensive properties in rats again in 1991. Several studies over the years have demonstrated that leaf, bark, root, stem, and seed extracts of graviola are antibacterial in vitro against numerous pathogens, and that the bark has antifungal properties. Graviola seeds demonstrated active antiparasitic properties in a 1991 study,8 and a leaf extract showed to be active against malaria in two other studies (in 1990 and 1993). Purdue University and/or its staff have filed at least nine U.S. and/or international patents on their work around the antitumorous and insecticidal properties and uses of these acetogenins. In one of their reviews, titled “Recent Advances in Annonaceous Acetogenins,” they state, “Recently, we reported that the Annonaceous acetogenins can selectively inhibit the growth of cancerous cells and also inhibit the growth of Adriamycin resistant tumor cells. As more acetogenins have been isolated and additional cytotoxicity assays have been conducted, we have noticed that, although most of acetogenins have high potencies among several solid human

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tumor cell lines, some of the derivatives within the different structural types and some positional isomers showed remarkable selectivity’s among certain cell lines; e.g., against prostate cancer (PC-3). We now understand the primary modes of action for the acetogenins. They are potent inhibitors of NADH: ubiquinone oxidoreductase, which is in an essential enzyme in complex I leading to oxidative phosphorylation in mitochondria. A recent report showed that they act directly at the ubiquinone-catalytic site(s) within complex I and in microbial glucose dehydrogenase. They also inhibit the ubiquinone-linked NADH oxidase that is peculiar to the plasma membranes of cancerous cells." 193BCancer research is ongoing on these important plants and plant chemicals, as several pharmaceutical companies, universities and scientists continue to research, test, patent, and attempt to synthesize these chemicals into new chemotherapeutic drugs. In addition, researchers have reported that NADH dehydrogenase inhibitors can suppress HIV infection. As this is a familiar property of Annonaceous acetogenins, several acetogenins found in graviola and other Annona plants have been submitted to the NIH anti-AIDS screening program by Purdue University; research work is continuing in this area as well. One researcher summarized his work eloquently: “At the time of preparation (August 1998) of this current review, over 350 Annonaceous acetogenins have been isolated from 37 species.” 194BGraviola has a lengthy history of safe use within traditional medical systems (over 5,000 years) as an herbal remedy for many conditions. Fruit and seeds of the Graviola plant are not recommended at this time. 195BGET WELL SOON: COOL, NATURAL PLANT CHEMICALS FOUND IN GRAVIOLA 196BThe Annonaceous acetogenins discovered in graviola thus far include: annocatalin, annohexocin, annomonicin, annomontacin, annomuricatin A & B, annomuricin A thru E, annomutacin, annonacin, annonacinone, annopentocin A thru C, cis-annonacin, ciscorossolone, cohibin A thru D, corepoxylone, coronin, corossolin, corossolone, donhexocin, epomuricenin A & B, gigantetrocin, gigantetrocin A & B, gigantetrocinone, gigantetronenin, goniothalamicin, iso-annonacin, javoricin, montanacin, montecristin, muracin A thru G, muricapentocin, muricatalicin, muricatalin, muri-catenol, muricatetrocin A & B muricatin D, muricatocin A thru C muricin H, muricin I, muricoreacin, murihexocin 3, murihexocin A thru C, murihexol, murisolin, robustocin, rolliniastatin 1 & 2, saba-delin, solamin, uvariamicin I & IV, xylomaticin. 197BActive components: Akaloids (Annonaceus Acetogenins), Muricoreacin, Munhexoin C, Mono-tetrahydrofuran, acetogenins, annomuricin E, Miricapetocin. 198BGET WELL SOON: FASCINATING RESEARCH ON GRAVIOLA

1. 199BThe Natural Health Bible for Dogs and Cats, Shawn Messonier, DVM

2. 200BWhole Dog Journal

3. 201BDr. Gary, Clinician’s Handbook of Natural Healing

4. 202BDr. James Duke, The Green Pharmacy

5. 203BNatural News

6. 204BDra. Hulda Clark, libro "The Cure for All Cancers" (best-selling book)

7. 205BPerdue Universidty (USA). Investigación sobre la acción inhibidora de derivados de Annonaceous en células cancerígenas. 1997.

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Anticancerous & Antitumor Actions: 1. Messonnier DVM, Shawn; The Natural Health Bible for Dogs and Cats Graviola for Dog Bone Cancer |

eHow.com

2. Kojima, N. “Systematic synthesis of antitumor Annonaceous acetogenins” Yakugaku Zasshi. 2004; 124(10): 673-81.

3. Tormo, J. R., et al. “In vitro antitumor structure-activity relationships of threo/trans/threo mono-tetrahydro-furanic acetogenins: Correlations with their inhibition of mitochondrial complex I.” Oncol. Res. 2003; 14(3): 147-54.

4. Yuan, S. S., et al. “Annonacin, a mono-tetrahydrofuran acetogenin, arrests cancer cells at the G1 phase and causes cytotoxicity in a Bax- and caspase-3-related pathway.” Life Sci. 2003 May: 72(25): 2853-61.

5. Liaw, C. C., et al. “New cytotoxic monotetrahydrofuran Annonaceous acetogenins from Annona muricata.” J. Nat. Prod. 2002; 65(4): 470-75

6. Gonzalez-Coloma, A., et al. “Selective action of acetogenin mitochondrial complex I inhibitors.” Z. Naturforsch. 2002; 57(11-12): 1028-34.

7. Chang, F. R., et al. “Novel cytotoxic Annonaceous acetogenins from Annona muricata.” J. Nat. Prod. 2001; 64(7): 925-31.

8. Jaramillo, M. C., et al. “Cytotoxicity and antileishmanial activity of Annona muricata pericarp.” Fitoterapia. 2000; 71 (2): 183-6.

9. Betancur-Galvis, L., et al. “Antitumor and antiviral activity of Colombian medicinal plant extracts.” Mem. Inst. Oswaldo Cruz. 1999; 94(4): 531-35.

10. Kim, G. S., et al. “Muricoreacin and murihexocin C, mono-tetrahydrofuran acetogenins, from the leaves of Annona muricata.” Phytochemistry. 1998; 49(2): 565-71.

11. Kim, G. S., et al. “Two new mono-tetrahydrofuran ring acetogenins, annomuricin E and muricapentocin, from the leaves of Annona muricata.” J. Nat. Prod. 1998; 61(4): 432-36.

12. Nicolas, H., et al. “Structure-activity relationships of diverse Annonaceous acetogenins against multidrug resistant human mammary adenocarcinoma (MCF-7/Adr) cells.” J. Med. Chem. 1997; 40(13): 2102-6.

13. Zeng, L., et al. “Five new monotetrahydrofuran ring acetogenins from the leaves of Annona muricata.” J. Nat. Prod. 1996; 59(11): 1035-42.

14. Wu, F. E., et al. “Two new cytotoxic monotetrahydrofuran Annonaceous acetogenins, annomuricins A and B, from the leaves of Annona muricata.” J. Nat. Prod. 1995; 58(6): 830-36.

15. Oberlies, N. H., et al. “Tumor cell growth inhibition by several Annonaceous acetogenins in an in vitro disk diffusion assay.” Cancer Lett. 1995; 96(1): 55-62.

16. Wu, F. E., et al. “Additional bioactive acetogenins, annomutacin and (2,4-trans and cis)-10R-annonacin-A-ones, from the leaves of Annona muricata.” J. Nat. Prod. 1995; 58(9): 1430-37.

17. Wu, F. E., et al. “New bioactive monotetrahydrofuran Annonaceous acetogenins, annomuricin C and muricatocin C, from the leaves of Annona muricata.” J. Nat. Prod. 1995; 58(6): 909-5.

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18. 224BWu, F. E., et al. “Muricatocins A and B, two new bioactive monotetrahydrofuran Annonaceous acetogenins from the leaves of Annona muricata.” J. Nat. Prod. 1995; 58(6): 902-8.

19. 225BSundarrao, K., et al. “Preliminary screening of antibacterial and antitumor activities of Papua New Guinean native medicinal plants.” Int. J. Pharmacog. 1993; 31(1): 3-6.

226BAntimicrobial Actions:

1. 227BTakahashi, J.A., et al. “Antibacterial activity of eight Brazilian Annonaceae plants.” Nat.Prod. Res. 2006; 20(1): 21-6.

2. 228BBetancur-Galvis, L., et al. “Antitumor and antiviral activity of Colombian medicinal plant extracts.” Mem. Inst. Oswaldo Cruz 1999; 94(4): 531-35.

3. 229BAntoun, M. D., et al. "Evaluation of the flora of Puerto Rico for in vitro cytotoxic and anti-HIV activities." Pharmaceutical Biol. 1999; 37(4): 277-280.

4. 230BPadma, P., et al. “Effect of the extract of Annona muricata and Petunia nyctaginiflora on Herpes simplex virus.” J. Ethnopharmacol. 1998; 61(1): 81–3.

5. 231BSundarrao, K., et al. “Preliminary screening of antibacterial and antitumor activities of Papua New Guinean native medicinal plants.” Int. J. Pharmacog. 1993; 31(1): 3–6.

6. 232BMisas, C. A. J., et al. “Contribution to the biological evaluation of Cuban plants. IV.” Rev. Cubana Med. Trop. 1979; 31(1): 29–35.

233BAntidepressant & Antistress Actions:

1. 234BPadma, P., et al. “Effect of Annona muricata and Polyalthia cerasoides on brain neurotransmitters and enzyme monoamine oxidase following cold immobilization stress.” J. Natural Remedies 2001; 1(2): 144–46.

2. 235BHasrat, J. A., et al. “Screening of medicinal plants from Suriname for 5-HT 1A ligands: Bioactive isoquinoline alkaloids from the fruit of Annona muricata.” Phytomedicine. 1997; 4(20: 133-140.

3. 236BPadma, P., et al. “Effect of alcohol extract of Annona muricata on cold immobilization stress induced tissue lipid peroxidation.” Phytother. Res. 1997; 11(4): 326-327.

4. 0BHasrat, J. A., et al. “Isoquinoline derivatives isolated from the fruit of Annona muricata as 5-HTergic 5-HT1A receptor agonists in rats: unexploited antidepressive (lead) products.” J. Pharm. Pharmacol. 1997; 49(11): 1145–49.

237BAntiparasitic, Antimalarial, & Insecticidal Actions:

1. 238BLuna, J. S., et al. “Acetogenins in Annona muricata L. (Annonaceae) leaves are potent molluscicides.” Nat. Prod. Res. 2006; 20(3): 253-7.

2. 239BJaramillo, M. C., et al. “Cytotoxicity and antileishmanial activity of Annona muricata pericarp.” Fitoterapia. 2000; 71(2): 183–6.

3. 240BAlali, F. Q., et al. “Annonaceous acetogenins as natural pesticides; potent toxicity against insecticide-susceptible and resistant German cockroaches (Dictyoptera: Blattellidae).” J. Econ. Entomol. 1998; 91(3): 641-9.

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4. Antoun, M. D., et al. "Screening of the flora of Puerto Rico for potential antimalarial bioactives.” Int. J. Pharmacog. 1993; 31(4): 255–58.

5. Heinrich, M., et al. “Parasitological and microbiological evaluation of Mixe Indian medicinal plants (Mexico).” J. Ethnopharmacol. 1992; 36(1): 81–5.

6. Bories, C., et al. “Antiparasitic activity of Annona muricata and Annona cherimolia seeds.” Planta Med. 1991; 57(5): 434–36.

7. Gbeassor, M., et al. “In vitro antimalarial activity of six medicinal plants.” Phytother. Res. 1990; 4(3): 115–17.

8. Tattersfield, F., et al. “The insecticidal properties of certain species of Annona and an Indian strain of Mundulea sericea (Supli).” Ann. Appl. Biol. 1940; 27: 262–73.

Anticonvulsant, Antispasmodic, & Smooth Muscle Relaxant Actions:

1. N’gouemo, P., et al. “Effects of ethanol extract of Annona muricata on pentylenetetrazolinduced convulsive seizures in mice.” Phytother. Res. 1997; 11(3): 243–45.

2. Feng, P. C., et al. “Pharmacological screening of some West Indian medicinal plants.” J. Pharm. Pharmacol. 1962; 14: 556–61.

Hypotensive & Cardiodepressant Actions

• Carbajal, D., et al. “Pharmacological screening of plant decoctions commonly used in Cuban folk medicine.” J. Ethnopharmacol. 1991; 33(1/2): 21–4.

• Feng, P. C., et al. “Pharmacological screening of some West Indian medicinal plants.” J. Pharm. Pharmacol. 1962; 14: 556–61.

• Meyer, T. M. “The alkaloids of Annona muricata.” Ing. Ned. Indie. 1941; 8(6): 64. References

1. Feng, P. C., et al. “Pharmacological screening of some West Indian medicinal plants.” J. Pharm. Pharmacol. 1962; 14: 556–61.

2. Meyer, T. M. “The alkaloids of Annona muricata.” Ing. Ned. Indie. 1941; 8(6): 64.

3. Carbajal, D., et al. “Pharmacological screening of plant decoctions commonly used in Cuban folk medicine.” J. Ethnopharmacol. 1991; 33(1/2): 21–4.

4. Misas, C. A. J., et al. “Contribution to the biological evaluation of Cuban plants. IV.” Rev. Cubana Med. Trop. 1979; 31(1): 29–35.

5. Sundarrao, K., et al. “Preliminary screening of antibacterial and antitumor activities of Papua New Guinean native medicinal plants.” Int. J. Pharmacog. 1993; 31(1): 3–6.

6. Heinrich, M., et al. “Parasitological and microbiological evaluation of Mixe Indian medicinal plants (Mexico).” J. Ethnopharmacol. 1992; 36(1): 81–5.

7. Lopez, Abraham A. M. “Plant extracts with cytostatic properties growing in Cuba. I.” Rev. Cubana Med. Trop. 1979; 31(2): 97–104.

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8. 261BBories, C., et al. “Antiparasitic activity of Annona muricata and Annona cherimolia seeds.” Planta Med. 1991; 57(5): 434–36.

9. 262BAntoun, M. D., et al. "Screening of the flora of Puerto Rico for potential antimalarial bioactives.” Int. J. Pharmacog. 1993; 31(4): 255–58.

10. 263BGbeassor, M., et al. “In vitro antimalarial activity of six medicinal plants.” Phytother. Res. 1990; 4(3): 115–17.

11. 264BTattersfield, F., et al. “The insecticidal properties of certain species of Annona and an Indian strain of Mundulea sericea (Supli).” Ann. Appl. B iol. 1940; 27: 262–73.

12. 265BHasrat, J. A., et al. “Isoquinoline derivatives isolated from the fruit of Annona muricata as 5- HTergic 5-HT1A receptor agonists in rats: unexploited antidepressive (lead) products.” J. Pharm. Pharmacol. 1997; 49(11): 1145–49.

13. 266BAnon. Unpublished data, National Cancer Institute. Nat Cancer Inst Central Files (1976). From NAPRALERT Files, University of Illinois, 1995.

14. 267BZeng, L., et al. “Five new monotetrahydrofuran ring acetogenins from the leaves of Annona muricata.” J. Nat. Prod. 1996; 59(11): 1035–42.

15. 268BRieser, M. J., et al. “Five novel mono-tetrahydrofuran ring acetogenins from the seeds of Annona muricata.” J. Nat. Prod. 1996; 59(2): 100–8.

16. 269BWu, F. E., et al. “Additional bioactive acetogenins, annomutacin and (2,4-trans and cis)-10Rannonacin- A-ones, from the leaves of Annona muricata.” J. Nat. Prod. 1995; 58(9): 1430–37.

17. 270BWu, F. E., et al. “New bioactive monotetrahydrofuran Annonaceous acetogenins, annomuricin C and muricatocin C, from the leaves of Annona muricata.” J. Nat. Prod. 1995; 58(6): 909–15.

18. 271BWu, F. E., et al. “Muricatocins A and B, two new bioactive monotetrahydrofuran Annonaceous acetogenins from the leaves of Annona muricata.” J. Nat. Prod. 1995; 58(6): 902–8.

19. 272BWu, F. E., et al. “Two new cytotoxic monotetrahydrofuran Annonaceous acetogenins, annomuricins A and B, from the leaves of Annona muricata.” J. Nat. Prod. 1995; 58(6): 830–36.

20. 273BRieser, M. J., et al. “Bioactive single-ring acetogenins from seed extracts of Annona muricata.” Planta Med. 1993; 59(1): 91–2.

21. 274BRieser, M. J., et al. “Muricatacin: a simple biologically active acetogenin derivative from the seeds of Annona muricata (Annonaceae)” Tetrahedron Lett. 1991; 32(9): 1137– 40.

22. 275BKim, G. S., et al. “Muricoreacin and murihexocin C, mono-tetrahydrofuran acetogenins, from the leaves of Annona muricata. Phytochemistry 1998; 49(2): 565–71.

23. 276BPadma, P., et al. “Effect of the extract of Annona muricata and Petunia nyctaginiflora on Herpes simplex virus. J. Ethnopharmacol. 1998; 61(1): 81–3.

24. 277BGleye, C., et al. “Cis-monotetrahydrofuran acetogenins from the roots of Annona muricata 1. J. Nat. Prod. 1998; 61(5): 576–9.

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25. Kim, G. S., et al. “Two new mono-tetrahydrofuran ring acetogenins, annomuricin E and muricapentocin, from the leaves of Annona muricata.” J. Nat. Prod. 1998; 61(4): 432–36.

26. Liaw, C. C., et al. “New cytotoxic monotetrahydrofuran Annonaceous acetogenins from Annona muricata.” J. Nat. Prod. 2002; 65(4): 470–75.

27. Chang, F. R., et al. “Novel cytotoxic annonaceous acetogenins from Annona muricata.” J. Nat. Prod. 2001; 64(7): 925–31.

28. Jaramillo, M. C., et al. “Cytotoxicity and antileishmanial activity of Annona muricata pericarp.” Fitoterapia 2000; 71(2): 183–6.

29. Betancur-Galvis, L., et al. “Antitumor and antiviral activity of Colombian medicinal plant extracts.” Mem. Inst. Oswaldo Cruz 1999; 94(4): 531-35.

30. Nicolas, H., et al. “Structure-activity relationships of diverse Annonaceous acetogenins against multidrug resistant human mammary adenocarcinoma (MCF-7/Adr) cells.” J. Med. Chem. 1997; 40(13): 2102–6.

31. Keinan, E., et al. “Antibody-catalyzed organic and organometallic transformations and chemical libraries of Annonaceous acetogenins.” The Skaggs Institute for Chemical Biology Scientific Report 1997–1998.

32. Zeng, L., et al., “Recent advances in Annonaceous acetogenins.” Nat. Prod Rep. 1996; 13(4): 275–306.

33. Anon., Purdue News September 1997; Purdue University, West Lafayette, IN. http://www.purdue.edu/UNS/newsandphotos.html

34. Feras, Q., et al. “Annonaceous acetogenins: Recent progress.” J. Nat. Prod. 1999; 62(3): 504- 540.

35. Wang, L. Q., et al. “Annonaceous acetogenins from the leaves of Annona montana.” Bioorg. Med. Chem. 2002; 10(3): 561-65.

36. Padma, P., et al. “Effect of Annona muricata and Polyalthia cerasoides on brain neurotransmitters and enzyme monoamine oxidase following cold immobilization stress.” J. Natural Remedies 2001; 1(2): 144–46.

37. N’gouemo, P., et al. “Effects of ethanol extract of Annona muricata on pentylenetetrazol-induced convulsive seizures in mice.” Phytother. Res. 1997; 11(3): 243–45.

More References: Graviola (Annona muricata) Takahashi, J. A., et al. "Antibacterial activity of eight Brazilian Annonaceae plants." Nat. Prod. Res. 2006; 20(1): 21-6. Kojima, N. "Systematic synthesis of antitumor Annonaceous acetogenins" Yakugaku Zasshi. 2004; 124(10): 673- 81 Yuan, S. S., et al. "Annonacin, a mono-tetrahydrofuran acetogenin, arrests cancer cells at the G1 phase and causes cytotoxicity in a Bax- and caspase-3-related pathway." Life Sci. 2003 May: 72(25): 2853-61. Liaw, C. C., et al. "New cytotoxic monotetrahydrofuran Annonaceous acetogenins from Annona muricata." J. Nat. Prod. 2002; 65(4): 470-75. Gonzalez-Coloma, A., et al. "Selective action of acetogenin mitochondrial complex I inhibitors." Z. Naturforsch. 2002; 57(11-12): 1028-34. Chang, F. R., et al. "Novel cytotoxic Annonaceous acetogenins from Annona muricata." J. Nat. Prod. 2001; 64(7): 925-31. Padma, P., et al. "Effect of Annona muricata and Polyalthia cerasoides on brain neurotransmitters and enzyme monoamine oxidase following cold immobilization stress." J. Natural Remedies 2001; 1(2): 144-46. Jaramillo, M. C., et al. "Cytotoxicity and antileishmanial activity of Annona muricata

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pericarp." Fitoterapia. 2000; 71 (2): 183-6. Betancur-Galvis, L., et al. "Antitumor and antiviral activity of Colombian medicinal plant extracts." Mem. Inst. Oswaldo Cruz. 1999; 94(4): 531-35. Kim, G. S., et al. "Muricoreacin and murihexocin C, mono-tetrahydrofuran acetogenins, from the leaves of Annona muricata." Phytochemistry. 1998; 49(2): 565-71. Kim, G. S., et al. "Two new mono-tetrahydrofuran ring acetogenins, annomuricin E and muricapentocin, from the leaves of Annona muricata." J. Nat. Prod. 1998; 61(4): 432-36. Nicolas, H., et al. "Structure-activity relationships of diverse Annonaceous acetogenins against multidrug resistant human mammary adenocarcinoma (MCF-7/Adr) cells." J. Med. Chem. 1997; 40(13): 2102-6. Zeng, L., et al. "Five new monotetrahydrofuran ring acetogenins from the leaves of Annona muricata." J. Nat. Prod. 1996; 59(11): 1035-42. Wu, F. E., et al. "Two new cytotoxic monotetrahydrofuran Annonaceous acetogenins, annomuricins A and B, from the leaves of Annona muricata." J. Nat. Prod. 1995; 58(6): 830-36. Wu, F. E., et al. "Additional bioactive acetogenins, annomutacin and (2,4-trans and cis)-10R-annonacin- A-ones, from the leaves of Annona muricata." J. Nat. Prod. 1995; 58(9): 1430-37. Wu, F. E., et al. "New bioactive monotetrahydrofuran Annonaceous acetogenins, annomuricin C and muricatocin C, from the leaves of Annona muricata." J. Nat. Prod. 1995; 58(6): 909- 5. Wu, F. E., et al. "Muricatocins A and B, two new bioactive monotetrahydrofuran Annonaceous acetogenins from the leaves of Annona muricata." J. Nat. Prod. 1995; 58(6): 902-8. Sundarrao, K., et al. "Preliminary screening of antibacterial and antitumor activities of Papua New Guinean native medicinal plants." Int. J. Pharmacog. 1993; 31(1): 3-6. Heinrich, M., et al. "Parasitological and microbiological evaluation of Mixe Indian medicinal plants (Mexico)." J. Ethnopharmacol. 1992; 36(1): 81-5. Mountain Graviola (Annona montana) Liaw, C. C., et al. "Novel cytotoxic monotetrahydrofuranic Annonaceous acetogenins from Annona montana." Bioorg. Med. Chem. 2005 Aug; 13(15): 4767-76. Tormo, J. R., et al. "In vitro antitumor structure-activity relationships of threo/trans/threo/trans/erythro bis-tetrahydrofuranic acetogenins: correlations with their inhibition of mitochondrial complex I." Oncol. Res. 2005; 15(3): 129-38. Liaw, C. C., et al. "Montacin and cis-montacin, two new cytotoxic monotetrahydrofuran Annonaceous acetogenins from Annona montana." J. Nat. Prod. 2004; 67(11): 1804-8. Liaw, C. C., et al. "Nine new cytotoxic monotetrahydrofuranic Annonaceous acetogenins from Annona montana." Planta Med. 2004; 70(10): 948-59. Kojima, N. "Systematic synthesis of antitumor annonaceous acetogenins." Yakugaku Zasshi. 2004; 124(10): 673-81 Wang, L. Q., et al. "Annonaceous acetogenins from the leaves of Annona montana." Bioorg. Med. Chem. 2002; 10(3): 561-5. Wang, L. Q., et al. "Cytotoxic mono-tetrahydrofuran ring acetogenins from leaves of Annona montana." Planta Med. 2001 Dec; 67(9): 847-52. Mootoo, B. S., et al. "Three novel monotetrahydrofuran annonaceous acetogenins from Annona montana."J. Nat. Prod. 2000; 63(6): 807-11. Yang, S., et al. "Chemical constituents of Annonaceae plants and their antitumor activities." Zhongguo Yi. Xue. Ke. Xue. Yuan. Xue. Bao. 2000 Aug; 22(4): 376-82. Wang, L. Q., et al. "Four monotetrahydrofuran ring acetogenins, montanacins B-E, from Annona montana." Chem. Pharm. Bull. 2000; 48(8): 1109-13. Wu, Y. C., et al. "Bioactive constitutents from the stems of Annona montana." Planta Med. 1995 Apr; 61(2): 146-9. Jossang, A., et al. "Annomonysvin: a new cytotoxic gamma-lactone-monotetrahydrofuranyl acetogenin from Annona montana." J. Nat. Prod. 1991 Jul-Aug; 54(4): 967-71. 293BMedicinal plants and cancer chemoprevention: 294BDesai AG, Qazi GN, Ganju RK, El-Tamer M, Singh J, Saxena AK, Bedi YS, 295BTaneja SC, Bhat HK. 296BDepartment of Environmental Health Sciences, Columbia University 297BCancer is the second leading cause of death worldwide. Although great advancements have been made in the treatment and control of cancer progression, significant deficiencies and room for improvement remain. A number of undesired side effects sometimes occur during chemotherapy. Natural therapies, such as the use of plant-derived products in cancer treatment, may reduce adverse side effects. Currently, a few plant products are being used to treat cancer. However, a myriad of many plant products exist that have shown very promising anti-cancer properties in vitro, but have yet to be evaluated in humans. Further study is required to determine the efficacy of these plant products in treating cancers in humans. This review will focus on the various plant-derived chemical compounds that have, in recent years, shown promise as anticancer agents and will outline their potential mechanism of action. 298BPMID: 18781909 [PubMed - indexed for MEDLINE]

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Related articles Garlic [Allium sativum]: a review of its potential use as an anti-cancer agent. Medicinal plants from Peru: a review of plants as potential agents against cancer. Anticancer Agents Med Chem. 2006 Sep; 6(5):429-44. [Anticancer Agents Med Chem. 2006] Advances in cancer therapy with plant based natural products. Curr Med Chem. 2001 Oct; 8(12):1467-86. [Curr Med Chem. 2001] Clinical Abstrasts J Nat Prod 2002 Apr;65(4):470-5 New cytotoxic monotetrahydrofuran annonaceous acetogenins from Annona muricata. Liaw, C. C., et al. Three new monotetrahydrofuran annonaceous acetogenins, muricin H (1), muricin I (2), and cis-annomontacin (3), along with five known acetogenins, annonacin, annonacinone, annomontacin, murisolin, and xylomaticin, were isolated from the seeds of Annona muricata. Additionally, two new monotetrahydrofuran annonaceous acetogenins, cis-corossolone (4) and annocatalin (5), together with four known ones, annonacin, annonacinone, solamin, and corossolone, were isolated from the leaves of this species. The structures of all new isolates were elucidated and characterized by spectral and chemical methods. These new acetogenins exhibited significant activity in in vitro cytotoxic assays against two human hepatoma cell lines, Hep G(2) and 2,2,15. Compound 5 showed a high selectivity toward the Hep 2,2,15 cell line. J Nat Prod 2001 Jul;64(7):925-31 Novel cytotoxic annonaceous acetogenins from Annona muricata. Chang, F. R., et al. Seven new annonaceous acetogenins, muricins A-G (1-7), as well as five known compounds, a mixture of muricatetrocin A (8) and muricatetrocin B (9), longifolicin (10), corossolin (11), and corossolone (12), were isolated from the seeds of Annona muricata. The structures of all isolates were elucidated and characterized by spectral and chemical methods. These acetogenins showed significantly selective in vitro cytotoxicities toward the human hepatoma cell lines Hep G(2) and 2,2,15. J Asian Nat Prod Res 2001;3(4):267-76 Annonaceous acetogenins of the seeds from Annona muricata. Li, D. Y., et al. Muricatenol (1) is a new C37 non-THF ring acetogenin with four hydroxyls and one isolated double bond in the long aliphatic chain. 2, 4-cis-Gigantetrocinone (2) and 2, 4- trans-gigantetrocinone (3) have been isolated as their acetates by preparative TLC. 2, 4- trans-Isoannonacin-10-one (4) and 2, 4-trans-isoannonacin (5) have been isolated as only 2, 4-trans-form for the first time (no cis-form). Also four known acetogenins, gigantetrocin-A (6), gigantetrocin-B (7), annomontacin (8), gigantetronenin (9) and a mixture of N-fatty acyl tryptamines have been isolated (10). Their structures have been established on the basis of spectral analyses. The CHCl3 fraction of the seeds showed strong antitumor activities. J Med Chem 2000 Dec 14;43(25):4793-800 Semisynthesis of antitumoral acetogenins: SAR of functionalized alkyl-chain bistetrahydrofuranic acetogenins, specific inhibitors of mitochondrial complex I. Gallardo, T., et al. The acetogenins of Annonaceae are known by their potent cytotoxic activity. In fact, they are promising candidatesas a new future generation of antitumoral drugs to fight against the current chemio-therapic resistant tumors. The main target enzyme of these compounds is complex I (NADH:ubiquinone oxidoreductase) of the mitochondrial respiratory chain, a key enzymatic complex of energy metabolism. In an attempt to characterize the relevant structural factor of the acetogenins that determines the inhibitory potency against this enzyme, we have prepared a series of bis-tetrahydrofuranic acetogenins with different functional groups along the alkyl chain.

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324BFitoterapia 2000 Apr;71(2):183-6 325BCytotoxicity and antileishmanial activity of Annona muricata pericarp. 326BJaramillo, M. C., et al. 327BHexane, ethyl acetate and methanol extracts of Annona muricata pericarp were tested in vitro against Leishmania braziliensis and L. panamensis promastigotes, and against cell line U-937. The ethyl acetate extract was more active than the other extracts and even of Glucantime used as reference substance. Its fractionation led to the isolation of three acetogenins--annonacin, annonacin A and annomuricin A. 40 328BMe Arch Pharm Res 1999 Oct;22(5):524-8 329Bcis-Annonacin and (2,4)-cis-and trans-isoannonacins: cytotoxic 330Bmonotetrahydrofuran annonaceous 331Bacetogenins from the seeds of Annona cherimolia. 332BWoo, M. H., et al. 333BDepartment of Pharmacy, College of Pharmacy, Catholic University of Taegu-Hyosung, Kyongsan, Korea. cis-Annonacin (1) and the mixture of (2,4)-cis-and trans-isoannonacins (2 and 3), three known mono-tetrahydrofuran annonaceous acetogenins, have been isolated from the seeds of Annona cherimolia by the use of the brine shrimp lethality test (BST) for bioactivity directed fractionation. Their structures were elucidated based on spectroscopic and chemical methods. 1 showed potent cytotoxicities in the brine shrimp lethality test (BST) and among six human solid tumor cell lines with notable selectivity for the pancreatic cell line (PaCa-2) at about 1,000 times the potency of adriamycin. The mixture of 2 and 3 is over 10,000 times cytotoxic as adriamycin in the pancreatic cell line (PaCa-2). All of the compounds are about 10 to 100 times as cytotoxic as adriamycin in the prostate cell line (PC-3). 334BInst Oswaldo Cruz 1999 Jul-Aug;94(4):531-5 335BAntitumor and antiviral activity of Colombian medicinal plant extracts. 336BBetancur-Galvis, L., et al 337BExtracts of nine species of plants traditionally used in Colombia for the treatment of a variety of diseases were tested in vitro for their potential antitumor (cytotoxicity) and antiherpetic activity. MTT (Tetrazolium blue) and Neutral Red colorimetric assays were used to evaluate the re-duction of viability of cell cultures in presence and absence of the extracts. MTT was also used to evaluate the effects of the extracts on the lytic activity of herpes simplex virus type 2 (HSV-2). The 50% cytotoxic concentration (CC50) and the 50% inhibitory concentration of the viral effect (EC50) for each extract were calculated by linear re-gression analysis. Extracts from Annona muricata, A. cherimolia and Rollinia membranacea, known for their cytotoxicity were used as pos-itive controls. Likewise, acyclovir and heparin were used as positive controls of antiherpetic activity. Methanolic extract from Annona sp. on Hep-2 cells presented a CC50 value at 72 hr of 49.6x10(3)mg/ml. Neither of the other extracts examined showed a significant cytotoxicity. The aqueous extract from Beta vulgaris, the ethanol extract from Callisia grasilis and the methanol extract Annona sp. showed some antiherpetic activity with acceptable therapeutic indexes (the ratio of CC50 to EC50). These species are good candidates for further activity-monitored fractionation to identify active principles. 338BChem Biol Interact 1999 Nov 1;122(3):171-83 339BSpecific interactions of monotetrahydrofuranic Annonaceous acetogenins as 340Binhibitors of mitochondrial 341Bcomplex I. 342BTormo, J. R., et al. 343BAnnonaceous acetogenins (ACG) are a wide group of cytotoxic compounds isolated from plants of the Annonaceae family. Some of them are promising candidates to be a future new generation of antitumor drugs due to the ability to inhibit the NADH:ubiquinone oxidoreductase of the respiratory chain (mitochondrial complex I), main gate of the energy production in the cell. ACG are currently being tested on standard

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antitumor trials although little is known about the structure activity relationship at the molecular level. On recent studies, the relevance of several parts of the molecule for the inhibitory potency has been evaluated. Phytochemistry 1998 Sep;49(2):565-71 Muricoreacin and murihexocin C, mono-tetrahydrofuran aceto-genins, from the leaves of Annona muricata. Kim, G. S., et al. Bioactivity-directed fractionation of the leaves of Annona muricata L.(Annonaceae) resulted in the isolation of two new Annonaceous acetogenins, muricoreacin (1) and murihexocin C (2). Compounds 1 and 2 showed significant cytotoxicities among six human tumor cell lines with selectivities to the prostate adenocarinoma (PC-3) and pancreatic carcinoma (PACA-2) cell lines. 41 J Nat Prod 1999 Mar;62(3):504-40 Annonaceous acetogenins: recent progress. Alali, F. Q., et al. The Annonaceous acetogenins are promising new antitumor and pesticidal agents that are found only in the plant family Annonaceae. Chemically, they are derivatives of longchain fatty acids. Biologically, they exhibit their potent bioactivities through depletion of ATP levels via inhibiting complex I of mitochondria and inhibiting the NADH oxidase of plasma membranes of tumor cells. Thus, they thwart ATP-driven resistance mechanisms. This review presents the progress made in the chemistry, biology, and development of these compounds since December 1995. J Nat Prod 1998 Apr;61(4):432-6 Two new mono-tetrahydrofuran ring acetogenins, annomuricin E and muricapentocin, from the leaves of Annona muricata. Kim, G.S., et al. Bioactivity-directed fractionation of the leaf extract of Annona muricata L. (Annonaceae) has resulted in the isolation of two new Annonaceous acetogenins, annomuricine (1) and muricapentocin (2). Compounds 1 and 2 are monotetrahydrofuran ring acetogenins bearing two flanking hydroxyl groups; however, each has three additional hydroxyl groups. Compound 1 has an erythro 1,2-diol, and 2 has a 1,5,9-triol moiety. Both 1 and 2 showed significant cytotoxicities against six types of human tumors, with selectivities to the pancreatic carcinoma (PACA-2) and colon adenocarcinoma (HT-29) cell lines. Ethnopharmacol 1998 May;61(1):81-3 Effect of the extract of Annona muricata and Petunia nyctaginiflora on Herpes simplex virus. Padma, P., et al. Annona muricata (Annonaceae) and Petunia nyctaginiflora (Solana-ceae) were screened for their activity against Herpes simplex virus-1 (HSV-1) and clinical isolate (obtained from the human keratitis lesion). We have looked at the ability of extract(s) to inhibit the cytopathic effect of HSV-1 on vero cells as indicative of anti-HSV-1 potential. The minimum inhibitory concentration of ethanolic extract of A. muricata and aqueous extract of P. nyctaginiflora was found to be 1 mg/ml. J Med Chem 1997 Jun 20;40(13):2102-6 Structure-activity relationships of diverse Annonaceous acetogenins against multidrug resistant human mammary adenocarcinoma (MCF-7/Adr) cells. Oberlies, N.H., et al. Fourteen structurally diverse Annonaceous acetogenins, representing the three main classes of bis-adjacent, bis-nonadjacent, and single-THF ring(s), were tested for their ability to inhibit the growth of adriamycin resistant human mammary adenocarcinoma (MCF-7/Adr) cells. This cell line is resistant to treatment with adriamycin,

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vincristine, and vinblastine and is, thus, multidrug resistant (MDR). Among a series of bis-adjacent THF ring acetogenins, those with the stereochemistry of threo-trans-threotrans- erythro (from C-15 to C-24) were the most potent with as much as 250 times the potency of adriamycin. A spacing of 13 carbons between the flanking hydroxyl of the THF ring system and the gamma-unsaturated lactone seems to be optimum with a spacing of 11 and 9 carbons being significantly less active. Several single-THF ring compounds were also quite potent with gigantetrocin A (11) being the most potent compound tested. The acetogenins may, thus, have chemotherapeutic potential, especially with regard to MDR tumors. 369BJ Nat Prod 1996 Feb;59(2):100-8 370BFive novel mono-tetrahydrofuran ring acetogenins from the seeds of Annona 371Bmuricata. 372BRieser, M. J., et al. 373BBioactivity-directed fractionation of the seeds of Annona muricata L. (Annonaceae) resulted in the isolation of five new compounds: cis-annonacin (1), cisannonacin- 10-one (2), cis-goniothalamicin (3), arianacin (4), and javoricin (5). Three of these (1-3) are among the first cis mono-tetrahydrofuran ring acetogenins to be reported. NMR analyses of published model synthetic compounds, prepared cyclized formal acetals, and prepared Mosher ester derivatives permitted the determinations of absolute stereochemistries. Bioassays of the pure compounds, in the brine shrimp test, for the inhibition of crown gall tumors, and in a panel of human solid tumor cell lines for cytotoxicity, 42 evaluated relative potencies. Compound 1 was selectively cytotoxic to colon adenocarcinoma cells (HT-29) in which it was 10,000 times the potency of adriamycin. 374BJ Nat Prod 1996 Nov;59(11):1035-42 375BFive new monotetrahydrofuran ring acetogenins from the leaves of Annona 376Bmuricata. 377BZeng, L., et al. 378BBioactivity-directed fractionation of the leaves of Annona muricata resulted in the isolation of annopentocins A (1), B (2), and C(3), and cis- and trans-annomuricin-D-ones (4, 5). Compounds 1-3 are the first acetogenins reported bearing a mono-tetrahydrofuran (THF) ring with one flanking hydroxyl, on the hydrocarbon side, and another hydroxyl, on the lactone side, that is one carbon away from the THF ring. Compounds 4 and 5 were obtained in a mixture and are new mono-THF ring acetogenins bearing two flanking hydroxyls and an erythro-diol located between the THF and the ketolactone rings. Compound 1 was selectively cytotoxic to pancreatic carcinoma cells (PACA-2), and 2 and 3 were selectively cytotoxic to lung carcinoma cells (A-549); the mixture of 4 and 5 was selectively cytotoxic for the lung (A-549), colon (HT-29), and pancreatic (PACA-2) cell lines with potencies equal to or exceeding those of Adriamycin. 379BJ Nat Prod 1995 Sep;58(9):1430-7 380BAdditional bioactive acetogenins, annomutacin and (2,4-trans and 381Bcis)-10R-annonacin-A-ones, from the leaves of Annona muricata. 382BWu, F. E., et al. 383BIn a continuation of our research on bioactive components from the leaves of Annona muricata, three novel monotetrahydrofuran Annonaceous acetogenins, namely, annomutacin [1], (2,4-trans)- 10R-annonacin-A-one [2], and (2,4-cis)-10R-annonacin-Aone [3], have been identified. Their structures were deduced by ms, nmr, ir, and uv spectral and chemical methods, and the absolute configurations were determined by Mosher ester methodology. A known bioactive amide, N-p-coumaroyl tyramine, was also found. Compound 1 and the mixture of compounds 2 and 3 showed selective cytotoxicities against the human A-549 lung tumor cell line. 384BCancer Lett 1995 Sep 4;96(1):55-62 385BTumor cell growth inhibition by several Annonaceous acetogenins in an in vitro disk 386Bdiffusion assay. 387BOberlies, N. H., et al.

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The cell inhibition activities of several Annonaceous acetogenins, covering the three major structural classes of bis-adjacent, bis-non-adjacent, and single tetrahydrofuran (THF) ring compounds and their respective ketolactone rearrangement products, were tested in an in vitro disk diffusion assay against three murine (P388, PO3, and M17/Adr) and two human (H8 and H125) cancerous cell lines as well as a non-cancerous immortalized rat GI epithelial cell line (I18). The results demonstrate a dose-dependent inhibition of cancerous cell growth, while non-cancerous cell growth is not inhibited by the same dosages. All of the acetogenins, irrespective of their various structural types, inhibit the growth of adriamycin resistant tumor cells and non-resistant tumor cells at the same levels of potency. These results show that the Annon-aceous acetogenins are an extremely potent class of compounds, and their inhibition of cell growth can be selective for cancerous cells and also effective for drug resistant cancer cells, while exhibiting only minimal toxicity to 'normal' non-cancerous cells. J Nat Prod 1995 Jun;58(6):909-15 New bioactive monotetrahydrofuran Annonaceous acetogenins, annomuricin C and muricatocin C, from the leaves of Annona muricata. Wu, F. E., et al. The leaves of Annona muricata have yielded two additional monotetrahydrofuran Annonaceous acetogenins, annomuricin C [1] and muricatocin C [2]. Compounds 1 and 2 each possess five hydroxyl groups; two hydroxyl groups are at the C-10/C-11 and C-10/C-12 positions in 1 and 2, respectively. The absolute configurations of 1 and 2, except for positions C-10 and C-11 or C-12, were determined by Mosher ester methodology. The C-10/C-11 and C-10/C-12 acetonides (1c, 2c) suggested relative stereochemistry and significantly enhanced the cytotoxicities against the A-549 human lung and the MCF-7 human beast solid tumor cell lines. One known monotetrahydrofuran acetogenin, gigantetronenin, not described previously from this plant, was also found. Chem Biol Interact 1999 Nov 1;122(3):171-83 Specific interactions of monotetrahydrofuranic Annonaceous acetogenins as inhibitors of mitochondrial complex I. Tormo, J. R., et al. Annonaceous acetogenins (ACG) are a wide group of cytotoxic compounds isolated from plants of the Annonaceae family. Some of them are promising candidates to be a future new generation of antitumor drugs due to the ability to inhibit the NADH:ubiquinone oxidoreductase of the respiratory chain (mitochondrial complex I), main gate of the energy production in the cell. ACG are currently being tested on standard antitumor trials although little is known about the structure activity relationship at the molecular level. On recent studies, the relevance of several parts of the molecule for the inhibitory potency has been evaluated. Phytochemistry 1998 Sep;49(2):565-71 Muricoreacin and murihexocin C, mono-tetrahydrofuran aceto-genins, from the leaves of Annona muricata. Kim, G. S., et al. Bioactivity-directed fractionation of the leaves of Annona muricata L.(Annonaceae) resulted in the isolation of two new Annonaceous acetogenins, muricoreacin (1) and murihexocin C (2). Compounds 1 and 2 showed significant cytotoxicities among six human tumor cell lines with selectivities to the prostate adenocarinoma (PC-3) and pancreatic carcinoma (PACA-2) cell lines. 41 J Nat Prod 1999 Mar;62(3):504-40 Annonaceous acetogenins: recent progress. Alali, F. Q., et al. The Annonaceous acetogenins are promising new antitumor and pesticidal agents that are found only in the plant family Annonaceae. Chemically, they are derivatives of longchain fatty acids. Biologically, they exhibit their potent bioactivities through depletion of ATP levels via inhibiting complex I of mitochondria and inhibiting the NADH oxidase of plasma membranes of tumor cells. Thus, they thwart ATP-driven resistance

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mechanisms. This review presents the progress made in the chemistry, biology, and development of these compounds since December 1995. 408BJ Nat Prod 1998 Apr;61(4):432-6 409BTwo new mono-tetrahydrofuran ring acetogenins, annomuricin E and 410Bmuricapentocin, from the leaves of Annona muricata. 411BKim, G.S., et al. 412BBioactivity-directed fractionation of the leaf extract of Annona muricata L. (Annonaceae) has resulted in the isolation of two new Annonaceous acetogenins, annomuricine (1) and muricapentocin (2). Compounds 1 and 2 are monotetrahydrofuran ring acetogenins bearing two flanking hydroxyl groups; however, each has three additional hydroxyl groups. Compound 1 has an erythro 1,2-diol, and 2 has a 1,5,9-triol moiety. Both 1 and 2 showed significant cytotoxicities against six types of human tumors, with selectivities to the pancreatic carcinoma (PACA-2) and colon adenocarcinoma (HT-29) cell lines. 413BEthnopharmacol 1998 May;61(1):81-3 414BEffect of the extract of Annona muricata and Petunia nyctaginiflora on Herpes 415Bsimplex virus. 416BPadma, P., et al. 417BAnnona muricata (Annonaceae) and Petunia nyctaginiflora (Solana-ceae) were screened for their activity against Herpes simplex virus-1 (HSV-1) and clinical isolate (obtained from the human keratitis lesion). We have looked at the ability of extract(s) to inhibit the cytopathic effect of HSV-1 on vero cells as indicative of anti-HSV-1 potential. The minimum inhibitory concentration of ethanolic extract of A. muricata and aqueous extract of P. nyctaginiflora was found to be 1 mg/ml. 418BJ Med Chem 1997 Jun 20;40(13):2102-6 419BStructure-activity relationships of diverse Annonaceous acetogenins against 420Bmultidrug resistant human mammary adenocarcinoma (MCF-7/Adr) cells. 421BOberlies, N.H., et al. 422BFourteen structurally diverse Annonaceous acetogenins, representing the three main classes of bis-adjacent, bis-nonadjacent, and single-THF ring(s), were tested for their ability to inhibit the growth of adriamycin resistant human mammary adenocarcinoma (MCF-7/Adr) cells. This cell line is resistant to treatment with adriamycin, vincristine, and vinblastine and is, thus, multidrug resistant (MDR). Among a series of bis-adjacent THF ring acetogenins, those with the stereochemistry of threo-trans-threotrans- erythro (from C-15 to C-24) were the most potent with as much as 250 times the potency of adriamycin. A spacing of 13 carbons between the flanking hydroxyl of the THF ring system and the gamma-unsaturated lactone seems to be optimum with a spacing of 11 and 9 carbons being significantly less active. Several single-THF ring compounds were also quite potent with gigantetrocin A (11) being the most potent compound tested. The acetogenins may, thus, have chemotherapeutic potential, especially with regard to MDR tumors. 423BJ Nat Prod 1996 Feb;59(2):100-8 424BFive novel mono-tetrahydrofuran ring acetogenins from the seeds of Annona 425Bmuricata. 426BRieser, M. J., et al. 427BBioactivity-directed fractionation of the seeds of Annona muricata L. (Annonaceae) resulted in the isolation of five new compounds: cis-annonacin (1), cis-annonacin-10-one (2), cis-goniothalamicin (3), arianacin (4), and javoricin (5). Three of these (1-3) are among the first cis mono-tetrahydrofuran ring acetogenins to be reported. NMR analyses of published model synthetic compounds, prepared cyclized formal acetals, and prepared Mosher ester derivatives permitted the determinations of absolute stereochemistries. Bioassays of the pure compounds, in the brine shrimp test, for the inhibition of crown gall tumors, and in a panel of human solid tumor cell lines for cytotoxicity, 42 evaluated relative potencies. Compound 1 was selectively cytotoxic to colon adenocarcinoma cells (HT-29) in which it was 10,000 times the potency of adriamycin.

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J Nat Prod 1996 Nov;59(11):1035-42 Five new monotetrahydrofuran ring acetogenins from the leaves of Annona muricata. Zeng, L., et al. Bioactivity-directed fractionation of the leaves of Annona muricata resulted in the isolation of annopentocins A (1), B (2), and C(3), and cis- and trans-annomuricin-D-ones (4, 5). Compounds 1-3 are the first acetogenins reported bearing a mono-tetrahydrofuran (THF) ring with one flanking hydroxyl, on the hydrocarbon side, and another hydroxyl, on the lactone side, that is one carbon away from the THF ring. Compounds 4 and 5 were obtained in a mixture and are new mono-THF ring acetogenins bearing two flanking hydroxyls and an erythro-diol located between the THF and the ketolactone rings. Compound 1 was selectively cytotoxic to pancreatic carcinoma cells (PACA-2), and 2 and 3 were selectively cytotoxic to lung carcinoma cells (A-549); the mixture of 4 and 5 was selectively cytotoxic for the lung (A-549), colon (HT-29), and pancreatic (PACA-2) cell lines with potencies equal to or exceeding those of Adriamycin. J Nat Prod 1995 Sep;58(9):1430-7 Additional bioactive acetogenins, annomutacin and (2,4-trans and cis)-10R-annonacin-A-ones, from the leaves of Annona muricata. Wu, F. E., et al. In a continuation of our research on bioactive components from the leaves of Annona muricata, three novel monotetrahydrofuran Annonaceous acetogenins, namely, annomutacin [1], (2,4-trans)- 10R-annonacin-A-one [2], and (2,4-cis)-10R-annonacin-Aone [3], have been identified. Their structures were deduced by ms, nmr, ir, and uv spectral and chemical methods, and the absolute configurations were determined by Mosher ester methodology. A known bioactive amide, N-p-coumaroyl tyramine, was also found. Compound 1 and the mixture of compounds 2 and 3 showed selective cytotoxicities against the human A-549 lung tumor cell line. Cancer Lett 1995 Sep 4;96(1):55-62 Tumor cell growth inhibition by several Annonaceous acetogenins in an in vitro disk diffusion assay. Oberlies, N. H., et al. The cell inhibition activities of several Annonaceous acetogenins, covering the three major structural classes of bis-adjacent, bis-non-adjacent, and single tetrahydrofuran (THF) ring compounds and their respective ketolactone rearrangement products, were tested in an in vitro disk diffusion assay against three murine (P388, PO3, and M17/Adr) and two human (H8 and H125) cancerous cell lines as well as a non-cancerous immortalized rat GI epithelial cell line (I18). The results demonstrate a dose-dependent inhibition of cancerous cell growth, while non-cancerous cell growth is not inhibited by the same dosages. All of the acetogenins, irrespective of their various structural types, inhibit the growth of adriamycin resistant tumor cells and non-resistant tumor cells at the same levels of potency. These results show that the Annon-aceous acetogenins are an extremely potent class of compounds, and their inhibition of cell growth can be selective for cancerous cells and also effective for drug resistant cancer cells, while exhibiting only minimal toxicity to 'normal' non-cancerous cells. J Nat Prod 1995 Jun;58(6):909-15 New bioactive monotetrahydrofuran Annonaceous acetogenins, annomuricin C and muricatocin C, from the leaves of Annona muricata. Wu, F. E., et al. The leaves of Annona muricata have yielded two additional monotetrahydrofuran Annonaceous acetogenins, annomuricin C [1] and muricatocin C [2]. Compounds 1 and 2 each possess five hydroxyl groups; two hydroxyl groups are at the C-10/C-11 and C- 10/C-12 positions in 1 and 2, respectively. The absolute configurations of 1 and 2, except for positions C-10 and C-11 or C-12, were determined by Mosher ester methodology. The C-10/C-11 and C-10/C-12 acetonides (1c, 2c) suggested relative stereochemistry and significantly enhanced the

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cytotoxicities against the A-549 human lung and the MCF-7 human beast solid tumor cell lines. One known monotetrahydrofuran acetogenin, gigantetronenin, not described previously from this plant, was also found. 43 448BJ Nat Prod 1995 Jun;58(6):902-8 449BMuricatocins A and B, two new bioactive monotetrahydrofuran Annonaceous 450Bacetogenins from the leaves of Annona muricata. 451BWu, F. E., et. al. 452BThe leaves of Annona muricata have yielded the novel monotetra-hydrofuran Annonaceous acetogenins, muricatocins A [1] and B [2]. Each compound possesses five hydroxyl groups, with two hydroxyl groups at the C-10 and C-12 positions. The absolute configurations of 1 and 2 (except for positions C-10 and C-12) were determined by Mosher ester methodology. The C-10, C-12 acetonides (1c, 2c) suggested relative stereochemistry and significantly enhanced cytotoxicity against the A-549 human lung tumor cell line. Three known monotetra-hydrofuran acetogenins, annonacin A, (2,4- trans)-isoannonacin, and (2,4-cis)-isoannonacin, were also found. 453BChem Biol Interact 1995 Oct 20;98(1):1-13 454BDetermination of structure-activity relationships of Annonaceous acetogenins by inhibition of oxygen uptake in rat liver mitochondria. Landolt, J.L., et al. A new group of natural compounds, the Annonaceous acetogenins, have recently been determined to inhibit ATP production at a similar site of action and higher levels of potency as rotenone, i.e., at NADH- ubiquinone oxido-reductase, complex I of the mitochondrial electron-transport chain. The acetogenins had earlier been determined to be pesticidal, antimalarial, antimicrobial, anti-parasitic, cytotoxic, and in vivo active as potentially new antitumor agents. In order to determine structural activity relationships (SARs) among these compounds, at the subcellular level, several available acetogenins have been tested. Data obtained, from the inhibition of oxygen consumption by rat liver mitochondria, demonstrated that all of the twenty acetogenins tested are active with IC50 values in the range of 15-800 nM/mg protein. The IC50 value of rotenone was 17 nM/mg protein. 455BJ Nat Prod 1991 Jul-Aug;54(4):967-71 456B[Annomonysvin: a new cytotoxic gamma-lactone- monotetrahydrofuranyl 457Bacetogenin from Annona montana] 458BJossang, A., et al. 459BThe structure of annomontacin [I], a novel monotetrayhydrofuran fatty acid gammalactone (acetogenin) isolated from the seeds of Amnona montana, was determined by spectral analysis. The cytotoxicities in vitro of annomontacin [I], annonacinone [2], and annonacin were measured against murine leukemia L1210, human breast adenocarcinoma MDA-MB231, and human breast carcinoma MCF7 cell lines and compared with adriamycin. 460BJ Nat Prod 1990 Mar-Apr; 53(2):237-78 461BAnnonaceous acetogenins: a review. 462BRupprecht, J. K., et al. 463BThe Annonaceous acetogenins are a series of apparently polyketide- derived fatty acid derivatives that possess tetrahydrofuran rings and a methylated gamma-lactone (sometimes rearranged to a methyl ketolactone) with various hydroxyl, acetoxyl, and/or ketoxyl groups along the hydrocarbon chain. They exhibit a broad range of potent biological activities (cytotoxicity, antitumor, antimalarial, antimicrobial, immunosuppressant, antifeedant, and pesticidal). The sources, isolation, chemistry, biogenesis, and biological actions of these compounds, published to date, are tabulated and discussed. Strategies for structural elucidation are reviewed, and structural revisions and refinements are suggested for some of the previously published compounds. A Reserch study held in Japan on March 2002 that was studying various acetogenins found in several species of plants inoculated mice with lung cancer cells. One third received nothing (the control group), one third received the chemotherapy drug adriamycin, and one third received the main graviola acetogenin, annonacin (at a dosage of 10 mg/kg). At the end of two weeks, five of the six in the untreated control group were still alive and lung

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tumor sizes were then measured. The adriamycin group showed a 54.6% reduction of tumor mass over the control group—but 50% of the animals had died from toxicity (three of six). The mice receiving annonacin were all still alive, and the tumors were inhibited by 57.9%—slightly better than adriamycin—and without toxicity. This led the researchers to summarize; “This suggested that annonacin was less toxic in mice. On considering the antitumor activity and toxicity, annonacin might be used as a lead to develop a potential anticancer agent. The Annonaceous acetogenins discovered in graviola thus far include: annocatalin, annohexocin, annomonicin, annomontacin, annomuricatin A & B, annomuricin A thru E, annomutacin, annonacin, annonacinone, annopentocin A thru C, cis-annonacin, ciscorossolone, cohibin A thru D, corepoxylone, coronin, corossolin, corossolone, donhexocin, epomuricenin A & B, gigantetrocin, gigantetrocin A & B, gigantetrocinone, gigantetronenin, goniothalamicin, iso-annonacin, javoricin, montanacin, montecristin, muracin A thru G, muricapentocin, muricatalicin, muricatalin, muri-catenol, muricatetrocin A & B muricatin D, muricatocin A thru C muricin H, muricin I, muricoreacin, murihexocin 3, murihexocin A thru C, murihexol, murisolin, robustocin, rolliniastatin 1 & 2, saba-delin, solamin, uvariamicin I & IV, xylomaticin the studies, published in two separate journal articles, show that the Graviola, also known as Paw paw, compounds not only are effective in killing tumors that have resistance to anti-cancer agents, but also seem to have a special affinity for such resistant cells. The findings were detailed in the journal Cancer Letters and the Journal of Medicinal Chemistry. References:

1. Kim, G. S., et al. “Two new mono-tetrahydrofuran ring acetogenins, annomuricin E and muricapentocin, from the leaves of Annona muricata.” J. Nat. Prod. 1998; 61(4): 432-36.

2. Nicolas, H., et al. “Structure-activity relationships of diverse Annonaceous acetogenins against multidrug resistant human mammary adenocarcinoma (MCF-7/Adr) cells.” J. Med. Chem. 1997; 40(13): 2102-6.

3. Zeng, L., et al. “Five new monotetrahydrofuran ring acetogenins from the leaves of

4. Annona muricata.” J. Nat. Prod. 1996; 59(11): 1035-42.

5. Wu, F. E., et al. “Two new cytotoxic monotetrahydrofuran Annonaceous acetogenins, annomuricins A and B, from the leaves of Annona muricata.Chang, F. R., et al. “Novel cytotoxic Annonaceous acetogenins from Annona muricata.” J. Nat. Prod. 2001; 64(7): 925-31.

6. Jaramillo, M. C., et al. “Cytotoxicity and antileishmanial activity of Annona muricata

7. pericarp.” Fitoterapia. 2000; 71 (2): 183-6.

8. Betancur-Galvis, L., et al. “Antitumor and antiviral activity of Colombian medicinal plant extracts.” Mem. Inst. Oswaldo Cruz. 1999; 94(4): 531-35.

9. Kim, G. S., et al. “Muricoreacin and murihexocin C, mono-tetrahydrofuran acetogenins, from the leaves of Annona muricata.” Phytochemistry. 1998; 49(2): 565-71.” J. Nat. Prod. 1995; 58(6): 830-36.

10. Oberlies, N. H., et al. “Tumor cell growth inhibition by several Annonaceous acetogenins in an in vitro disk diffusion assay.” Cancer Lett. 1995; 96(1): 55-62.

11. Wu, F. E., et al. “Additional bioactive acetogenins, annomutacin and (2,4-trans and cis)- 10R-annonacin-A-ones, from the leaves of Annona muricata.” J. Nat. Prod. 1995; 58(9): 1430-37.

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12. 481BWu, F. E., et al. “New bioactive monotetrahydrofuran Annonaceous acetogenins, annomuricin C and muricatocin C, from the leaves of Annona muricata.” J. Nat. Prod. 1995; 58(6): 909-5. suggested dose : 2 tabs three times a day after two weeks discontinue the treatment for (2) days then re-start again

482BPurdue News 483BSeptember 1997 Graviola, Pawpaw Tree are common names for: Annona muricata 484BPawpaw shows promise in fighting drug-resistant tumors 485BWEST LAFAYETTE, Ind. -- The pawpaw tree, which bears the largest fruit native to North America, may bear new fruit forscientists seeking ways to fight cancer. Purdue University researcher Jerry McLaughlin, working with doctoral student Nicholas Oberlies, has found compounds in the bark of the tree that have shown preliminary success in fighting some drug-resistant cancers.The studies, published in two separate journal articles this summer, show that the pawpaw compounds not only are effective in killing tumors that have proven resistant to anti-cancer agents, but also seem to have a special affinity for such resistantcells. 486BThe findings were detailed in the journal Cancer Letters and the Journal of Medicinal Chemistry. Though further studies are needed to pinpoint exactly how the pawpaw compounds work within the cancer cell, McLaughlin says their effect is to pull the plug on the energy-producing mechanisms in the cell. McLaughlin notes, however, that the effect on drug-resistant cells has been studied only in laboratory cultures and will require additional study in animals before it can be tested in humans."Multidrug-resistant cancer is hard to treat because the cancer cell has developed a mechanism to get around the anticancer agent," says McLaughlin, professor of pharmacognosy in Purdue's School of Pharmacy and Pharmacal Science. "Tumor cells that survive chemotherapy treatments often recover with increased resistance to the agent used in the originaltreatment program as well as to other related drugs." 487BSuch resistance can develop when surviving cancer cells develop one or more mechanisms to accelerate the removal of noxious substances, including anti-cancer drugs. 488BOne of the most common mechanisms used to circumvent the anti-cancer agents is to develop a "pump" that is capable of pushing anti-cancer agents out of the cell before they can kill it. These pumps are called P-glycoprotein mediated pumps and are named for the type of protein used to construct and operate them. Though all cells have the ability to develop such a pump, normal cells seldom do. Even in cancer cells, which do not respond normally to the body's control mechanisms, only a small percentage of cells develop this pumping mechanism. "If having this pump was such a good deal, all cells would have it. But all cells don't," McLaughlin says. "In a given population of cancer cells in a person, maybe only 2 percent of the cancer cells possess this pump. But it's those 2 percent of cancer cells that eventually grow and expand to create drug-resistant tumors." 489BOne of the tricks currently attempted in treating cancer patients is to flood the body with other compounds to keep the pump busy, and then administer high doses of an anticancer agent in hopes that some of it will be able to stay in long enough to kill the cancer cell. "But the high doses of the drugs required for this treatment often produce side effects, such as loss of blood pressure, so the patient often succumbs to the side effects of the treatment," McLaughlin says. Though this pump mechanism is efficient at eliminating most anti-cancer agents, McLaughlin, whose research group has identified more than 40 pawpaw compounds with anti-cancer properties, discovered a series of the compounds, called Annonaceous acetogenins, that were capable of killing cancer cells that employed this mechanism. He then designed a laboratory study to analyze the cytotoxic or cell-killing effects of one of the compounds, called bullatacin, on human mammary cancer cells. The study compared bullatacin's effects on standard, nonresistant cancer cells and on multidrug-resistant cells.In the June issue of Cancer Letters, the research team reported that bullatacin prefere intially killed the multidrugresistant cells by inhibiting the production of adenosine triphosphate, or ATP. 490BATP is a compound that works to release energy in acell and is essential to all cell processes. "A multidrug-resistant cell requires a tremendous amount of energy to run the pump and extrude things out of the cell," McLaughlin says. "By inhibiting ATP production, we're essentially pulling the plug on its energy

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source."Though the pawpaw compounds also inhibited ATP production in noncancerous cells and nonresistant cancer cells, those cells were not affected as dramatically, McLaughlin says. Pawpaw shows promise in fighting drug-resistant tumors file:///C:/HTML-Flies/Grav-Purdue-Study.html 2 of 2 2/8/2005 12:42 PM "Normal cells and standard cancer cells may be able to minimize the effects of this compound because they don't require the vast amounts of energy needed by the pumprunning cells," McLaughlin says. "The resistant cell is using its extra energy for this pump as well as to grow, so it is really taxed for energy. When we mess with the energy supply, it kills the cell." McLaughlin and his group then did a follow-up study to test a series of 14 structurally similar pawpaw compounds to determine the structural features that maximize this biological activity in multidrug-resistant cancer cells. The results were published in the June issue of the Journal of Medicinal Chemistry. "This study tells us how to maximize this activity, so we have a pretty good idea what compounds we'd like to try in animals with multidrug-resistant tumors," McLaughlin says. If proven effective in animals and humans, McLaughlin says, the compounds may be used to treat multidrug resistance in a variety of cancers, because many types of cancer cells develop resistance by employing a pump.The studies were funded by National Institutes of Health/National Cancer Institute, the Indiana Elks Cancer Research Fundand Purdue Research Foundation. Purdue has filed a patent on the use of the pawpaw compounds. Source: Jerry McLaughlin, (765) 494-1455; e-mail, jac@pharmacy.purdue.edu Writer: Susan Gaidos, (765) 494-2081; e-mail, susan_gaidos@uns.purdue.edu Purdue News Service: (765) 494-2096; e-mail, purduenews@uns.purdue.edu ABSTRACT: Cancer Letters 115 (1997) 73-79 The Annonaceous acetogenin bullatacin is cytotoxic against multidrug-resistant human mammary adenocarcinoma cells Nicholas H. Oberlies, Vicki L. Croy, Marietta L. Harrison, Jerry L. McLaughlinDepartment of Medicinal Chemistry and Molecular Pharmacology, School of Pharmacy and Pharmacal Sciences, Purdue University.Cytotoxic effects of the Annonaceous acetogenin, bullatacin, were studied in multidrug-resistant (MDR) human mammary adenocarcinoma (MCF- 7/Adr) cells vs. the parental non-resistant wild type (MCF-7/wt) cells. Bullatacin was effectively cytotoxic to the MCF-7/Adr cells while it was more cytostatic to the MCF- 7/wt cells. ATP depletion is the mode of action of the Annonaceous acetogenins, and these agents offer a special advantage in the chemotherapeutic treatment of MDR tumors that have ATP-dependent mechanisms. ABSTRACT: J. Med. Chem. 1997, 40, 2102- 2106 Structure-activity relationships of diverse Annonaceous acetogenins against multidrugresistant human mammary adenocarcinoma (MCF-7/Adr) cells Nicholas H. Oberlies, Ching-jer Chang, Jerry L. McLaughlin Department of Medicinal Chemistry and Molecular Pharmacology, School of Pharmacy and Pharmacal Sciences, Purdue University. Fourteen structurally diverse Annonaceous acetogenins, representing the three main classes of bis-adjacent, bis-nonadjacent, and single-THF ring(s), were tested for their ability to inhibit the growth of adriamycin-resistant human mammary adenocarcinoma (MCF-7/Adr) cells. This cell line is resistant to treatment with adriamycin, vincristine, and vinblastine and is, thus, multidrug-resistant (MDR). Among a series of bis-adjacent THF ring acetogenins, those with the stereochemistry of threotrans- threo-trans-erythro (from C-15 to C-24) were the most potent with as much as 250 times the potency of adriamycin. A spacing of 13 carbons between the flanking hydroxyl of the THF ring system and the gamma-unsaturated lactone seems to be optimum with a spacing of 11 and 9 carbons being significantly less active. Several single-THF ring compounds were also quite potent, with gigantetrocin A (11) being the most potent compound tested. The acetogenins may, thus, have chemotherapeutic potential, especially with regard to MDR tumors.

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495B2. MEMORIAL SLOAN KETTERING 496Bwww.mskcc.org 497BGRAVIOLA 498BScientific Name: 499BAnnona muricata. Family: Annonaceae 500BCommon Name: 501Bcherimoya, guanabana, soursop, sour sop, custard apple, brazilian paw paw 502BGraviola,guanábano, guanavana, guanaba, corossol épineux, huanaba, toge-banreisi, 503Bdurian benggala, nangka blanda, cachiman épineux 504BClinical Summary 505BDerived from a tree in the rain forests of Africa, South America, and Southeast Asia. The bark, leaves, root, and fruits have been used as traditional remedies in many countries. Extracts o f graviola have been shown to have antiviral (1), antiparasitic, antirheumatic, astringent, emetic (2), antileishmanial and cytotoxic (3) (4) effects. Graviola has also been shown to be effective against multidrug resistant cancer cells line (5) (4). There are no large scale studies in humans on the effects of graviola. Alkaloids extracted from graviola may cause neuronal dysfunction and degeneration leading to symptoms reminiscent of Parkinson's Disease (6). 506BPurported uses

• 507BCancer treatment • 508BHerpes • 509BInfections • 510BParasitic infections • 511BSedation

512BConstituents 513BAcetogenins, quinolines, isoquinolines, annopentocins and annomuricins (7). Coreximine and reticuline subfractions have been isolated from root bark (6). 514BMechanism of Action 515BAnnonaceous acetogenins, a phytochemical isolated from the leaves, bark and twigs of graviola, are thought to be the active ingredient. The ethanolic extract of Annona muricata was found to inhibit the Herpes simplex virus (1). The ethyl acetate extract was found to be more active than the methanolic or hexanolic extracts. Extracts were shown to be effective against Leishmania braziliensis, L. panamensis promastigotes and the cancer cell line U 937 (3) and hepatoma cell lines(8) in vitro. Extracts were also shown to be lethal to the fresh-water mollusk Biomphalaria glabrata, which acts as a host for the parasitic worm Schistosoma mansoni (2). Alkaloids from graviola have been shown to be detrimental to the survival of dopaminergic nerve cells in vitro. This may in turn lead to neuronal dysfunction and degeneration. Graviola-induced cell death was inhibited by the supplementation of glucose suggesting that cell death was caused by energy depletion (6). 516BGraviola extracts have also been shown to be effective against the growth of Adriamycin resistant human mammary adenocarcinoma (MCF-7/Adr) by blocking the cancer cell's access to ATP and by inhibiting the actions of plasma membrane glycoprotein (5). Graviola may also have antidepressive activity due to its ability to stimulate serotonine receptors (7). 517BPharmacokinetics 518BLittle is known about the absorption, distribution, metabolism or excretion of graviola.

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Adverse Reactions Graviola may cause movement disorders and myeloneuropathy with symptoms similar to Parkinson's disease (6). Literature Summary and Critique There are no large scale studies in humans on the effects of graviola. References

1. Padma P, Pramod NP, Thyagarajan SP, Khosa RL. Effect of the extract of Annona muricata and Petunia nyctaginiflora on Herpes simplex virus. J Ethnopharmacol. 1998;61:81-3.

2. dos Santos AF,.Sant'Ana AE. Molluscicidal properties of some species of Annona. Phytomedicine. 2001;8:115-20.

3. Jaramillo MC, Arango GJ, Gonzalez MC, Robledo SM, Velez ID. Cytotoxicity and antileishmanial activity of Annona muricata pericarp. Fitoterapia 2000;71:183-6.

4. Liaw CC, Chang FR, Lin CY, Chou CJ, Chiu HF, Wu MJ et al. New cytotoxic monotetrahydrofuran annonaceous acetogenins from Annona muricata. J Nat.Prod. 2002;65:470-5.

5. Oberlies NH, Chang CJ, McLaughlin JL. Structure-activity relationships of diverse Annonaceous acetogenins against multidrug resistant human mammary adenocarcinoma (MCF-7/Adr) cells. J Med Chem. 1997;40:2102-6.

6. Lannuzel A, Michel PP, Caparros-Lefebvre D, Abaul J, Hocquemiller R, Ruberg M.Toxicity of Annonaceae for dopaminergic neurons: potential role in atypical parkinsonism in Guadeloupe. Mov Disord. 2002;17:84-90.

7. Roman G. Tropical myeloneuropathies revisited. Curr.Opin.Neurol 1998;11:539- 44. 8. Chen JC, Tsai CC, Chen LD, Chen HH, Wang WC. Therapeutic effect of gypenoside on chronic liver

injury and fibrosis induced by CCl4 in rats. Am.J Chin Med. 2000;28:175-85. 9. Graviola.org. http://www.graviola.org/cgi/gpp.cgi?000001-000001-

000000&../../1graviola/parsed_pages/graviola-data.htm (Accessed July 15, 2008) PUBMED/MEDLINE RESEARCH: Ingredients: Graviola leaf and stem, mullaca whole herb, guacatonga leaf and stem, espinheira santa leaf, bitter melon whole herb, vassourinha leaf, mutamba bark, cat’s claw vine. Suggested Use: Take 3-4 capsules three times daily on an empty stomach. Contraindications:

• Not to be used during pregnancy or while breast-feeding. • Several ingredients in this formula have demonstrated hypotensive, vasodilator, and cardiodepressant

activities in animal studies. People with low blood pressure or those taking antihypertensive drugs should check with their physician before taking Graviola and monitor their blood pressure accordingly.

• Do not take in conjuction with CoQ10 and other supplements which increase cellular ATP and may reduce the benefits.

Drug Interactions: This product may potentiate antihypertensive and cardiac depressant drugs. OVERVIEW OF CLINICAL DOCUMENTATION Other Practitioner Observations: Several ingredients in this formula have demonstrated significant in vitro antimicrobial properties. Chronic or long-term use of this plant may lead to die-off of friendly bacteria in the digestive tract due to its antimicrobial

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properties. Supplementing the diet with probiotics and digestive enzymes is advisable if this product is used for longer than 30 days. 535BDrinking plenty of water (at least 8 glasses a day) is helpful to reduce Herxheimer reactions. 536BClinical Documentation and Research: 537BThis formulated product has not been the subject of any clinical research. Available thirdparty documentation and clinical research on each ingredient in this formula can be found at the Natura Herbs website. A partial listing of published research on these ingredients is shown below: 538BGraviola (Annona muricata)

1. 539BLiaw, C. C., et al. “New cytotoxic monotetrahydrofuran Annonaceous acetogenins from Annona muricata.” J. Nat. Prod. 2002; 65(4): 470–75.

2. 540BChang, F. R., et al. “Novel cytotoxic Annonaceous acetogenins from Annona muricata.” J. Nat. Prod. 2001; 64(7): 925–31.

3. 541BZeng, B. B., et al. “Studies on mimicry of naturally occurring Annonaceous acetogenins: Non-THF analogues leading to remarkable selective cytotoxicity against human tumor cells.” Chemistry. 2003 Jan 3; 9(1): 282-90.

4. 542BNicolas, H., et al. “Structure-activity relationships of diverse Annonaceous acetogenins against multidrug resistant human mammary adenocarcinoma (MCF-7/Adr) cells.” J. Med. Chem. 1997; 40(13): 2102–6.

5. 543BGonzalez-Coloma, A., et al. “Selective action of acetogenin mitochondrial complex I inhibitors.” Z. Naturforsch [C]. 2002; 57(11-12): 1028-34.

6. 544BTormo, J. R., et al. “Specific interactions of monotetrahydrofuranic annonaceous acetogenins as inhibitors of mitochondrial complex I.” Chem. Biol. Interact. 1999 Nov 1; 122(3): 171-83.

545BMullaca (Physalis angulata)

1. 546BChiang, H., et al. “Antitumor agent, physalin F from Physalis angulata L.” Anticancer Res. 1992; 12(3): 837–43.

2. 547BIsmail, N., et al. “A novel cytotoxic flavonoid glycoside from Physalis angulata.” Fitoterapia 2001 Aug. 72(6): 676–79.

3. 548BHayashi T, A cytotoxic flavone from Scoparia dulcis L. Chem Pharm Bull (Tokyo). 1988 Dec; 36(12): 4849-51.

4. 549BChiang, H. et al. “Inhibitory effects of physalin B and physalin F on various human leukemia cells in vitro.” Anticancer Res. 1992; 12(4): 1155–62.

5. 550BKawai, M., et al. “Cytotoxic activity of physalins and related compounds against HeLa cells.” Pharmazie. 2002; 57(5): 348–50.

551BGuacatonga (Casearia sylvestris)

1. 552BOberlies, N. H., et al. "Novel bioactive clerodane diterpenoids from the leaves and twigs of Casearia sylvestris." J. Nat. Prod.2002; 65(2): 95-99.

2. 553BItokawa, H., et al. "New antitumor principles, casearins A-F, for Casearia sylvestris Sw. (Flacourtiaceae)" Chem. Pharm. Bull. (Tokyo) 1990; 38(12): 3384-88.

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3. Itokawa, H., et al. "Antitumor principles from Casearia sylvestris Sw. (Flacourtiaceae), structure elucidation of new clerodane diterpenes by 2-D NMR spectroscopy." Chem. Pharm. Bull. (Tokyo) 1988 March; 36(4): 1585-88.

4. Morita, H., et al. "Structures and cytotoxic activity relationship of casearins, new clerodane diterpenes from Casearia sylvestris Sw." Chem. Pharm. Bull. (Tokyo) 1991 Dec; 39(3): 693-97.

5. Bolzani Vda S., et al. " Search for antifungal and anticancer compounds from native plant species of Cerrado and Atlantic Forest." An. Acad. Bras. Cienc. 1999; 71(2): 181-7.

Espinheira Santa (Maytenus ilicifolia)

1. Itokawa, H., et al. “Antitumor substances from South American plants.” Pharmacobio. Dyn. 1992; 15(1): S-2.

2. Arisawa, M., et al. “Cell growth inhibition of KB cells by plant extracts.” Natural Med. 1994; 48(4): 338–347.

3. Shirota, O., et al. “Cytotoxic aromatic triterpenes from Maytenus ilicifolia and Maytenus chuchuhuasca.” J. Nat. Prod. 1994; 57(12): 1675–81.

4. Ladino, C.A., et al. “Folate-maytansinoids: target-selective drugs of low molecular weight.” Int. J. Cancer. 1997 Dec 10; 73(6): 859-64.

Bitter Melon (Momordica charantia)

1. Claflin, A. J., et al. “Inhibition of growth and guanylate cyclase activity of an undifferentiated prostate adenocarcinoma by an extract of the balsam pear (Momordica charantia abbreviata).” Proc. Natl. Acad. Sci. 1978; 75(2): 989–93.

2. Terenzi, A., et al. “Anti-CD30 (BER=H2) immunotoxins containing the type-1 ribosomeinactivating proteins momordin and PAP-S (pokeweed antiviral protein from seeds) display powerful antitumor activity against CD30+ tumor cells in vitro and in SCID mice.” Br. J. Haematol. 1996; 92(4): 872–79.

3. Nagasawa, H., et al. “Effects of bitter melon (Momordica charantia) or ginger rhizome (Zingiber offifinale Rosc.) on spontaneous mammary tumorigenesis in SHN mice.” Am. J. Clin. Med. 2002; 30(2–3): 195–205.

4. Pongnikorn, S., et al. “Effect of bitter melon (Momordica charantia Linn) on level and function of natural killer cells in cervical cancer patients with radiotherapy.” J. Med. Assoc.Thai. 2003; 86(1): 61-8.

Vassourinha (Scoparia dulcis)

1. Nishino, H. “Antitumor-promoting activity of scopadulcic acid B, isolated from the medicinal plant Scoparia dulcis L.” Oncology 1993; 50(2): 100–3.

2. Noda, Y., et al. “Enhanced cytotoxicity of some triterpenes toward leukemia L1210 cells cultured in low pH media; possibility of a new mode of cell killing.” Chem. Pharm. Bull. 1997; 45(10): 1665–70.

3. Zuco, V., et al. “Selective cytotoxicity of betulinic acid on tumor cell lines, but not on normal cells.” Cancer Lett. 2002; 175(1): 17–25.

4. Ahsan, M., et al. “Cytotoxic diterpenes from Scoparia dulcis.” J. Nat. Prod. 2003; 66(7): 958-61. Mutamba (Guazuma ulmifolia)

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1. 573BKashiwada, Y., et al. “Antitumor agents, 129. Tannins and related compounds as selective cytotoxic agents.” J. Nat. Prod. 1992; 55(8): 1033–43.

2. 574BIto, H., et al. “Antitumor activity of compounds isolated from leaves of Eriobotrya japonica.” J. Agric. Food Chem. 2002; 50(8): 2400–3.

575BCat’s Claw (Uncaria tomentosa)

1. 576BSheng, Y. “Induction of apoptosis and inhibition of proliferation in human tumor cells treated with extracts of Uncaria tomentosa.” Anticancer Res. 1998 Sep-Oct;18(5A): 3363-8.

2. 577BLemaire, I., et al. “Stimulation of interleukin-1 and -6 production in alveolar macrophages by the neotropical liana, Uncaria tomentosa.” J. Ethnopharmacol. 1999 Feb;64(2):109-15.

578BIndian J Exp Biol. 2007 May;45(5):480-5. 579B1: Baskar R, Rajeswari V, Kumar TS. 580BDepartment of Biotechnology, Kumaraguru College of Technology, Coimbatore 641 006, India Antioxidant potential of leaves of three different species of Annona was studied by using different in vitro models eg., 1,1-diphenyl-2-picryl hydrazyl (DPPH), 2,2-azinobis-(3-ethylbenzothizoline-6-sulphonate) (ABTS), nitric oxide, superoxide, hydroxy radical and lipid peroxidation. The ethanolic extract of A. muricata at 500 microg/ml showed maximum scavenging activity (90.05%) of ABTS radical cation followed by the scavenging of hydroxyl radical (85.88%) and nitric oxide (72.60%) at the same concentration. However, the extract showed only moderate lipid peroxidation inhibition activity. In contrast, the extract of A. reticulata showed better activity in quenching DPPH (89.37%) and superoxide radical (80.88%) respectively. A.squamosa extract exhibited least inhibition in all in vitro antioxidant models excepting hydroxyl radical (79.79%). These findings suggest that the extracts of A. muricata possess potent in vitro antioxidant activity as compared to leaves of A. squamosa and A. reticulata suggesting its role as an effective free radical scavenger, augmenting its therapeutic 581BPMID: 17569293 [PubMed - indexed for MEDLINE] 582BRelated articles

• 583B_ In vitro antioxidant studies of Annona squamosa Linn. leaves. Indian J Exp Biol. 2004 Aug; 42(8):803-7. [Indian J Exp Biol. 2004]

• 584B_ In vitro antioxidant studies of Sphaeranthus indicus (Linn). Indian J Exp Biol. 2006 Dec; 44(12):993-6. [Indian J Exp Biol. 2006]

• 585B_ In vitro antioxidant activity of Diospyros malabarica Kostel bark. Indian J Exp Biol. 2006 Jan; 44(1):39-44. [Indian J Exp Biol. 2006]

• 586B_ Cytisus scoparius link--a natural antioxidant. BMC Complement Altern Med. 2006 Mar 16; 6:8. Epub 2006 Mar 16. [BMC Complement Altern Med. 2006]

• 587B_ Antioxidant activity of Caesalpinia digyna root. J Ethnopharmacol. 2007 Sep 5; 113(2):284-91. Epub 2007 Jun 26. [J Ethnopharmacol. 2007]

• 588B_ J Ethnopharmacol. 2007 May 22: 111(3):630-5. Epub 2007 Jan 18. 589BAntiprotozoal and cytotoxic activities in vitro of Colombian Annonaceae. 590BOsorio E, Arango GJ, Jiménez N, Alzate F, Ruiz G, Gutiérrez D, Paco MA, 591BGiménez A, Robledo S. 592BGrupo de Investigación en Sustancias Bioactivas (GISB), Facultad de Química Farmacéutica, Corporación de Patologías Tropicales, Universidad de Antioquia, A.A. 1226, Medellín, Colombia. Ethnobotanical and chemotaxonomical studies for antiparasitic activity of Colombian Annonaceae were carried out. In vitro antiprotozoal activity of 36 extracts obtained from six different species was determined against promastigotes of three Leishmania species, epimastigotes of Trypanosoma cruzi and both chloroquine sensitive (F32) and resistant (W2) Plasmodium falciparum. Cytotoxic activity was evaluated in U-937 cells. Active extracts were selected according their selectivity index (SI). Extracts from Annona muricata, Rollinia exsucca, Rollinia

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pittieri and Xylopia aromatica were active against Leishmania spp. and Trypanosoma cruzi showing IC50 values lower than 25 microg/ml. Hexane extract from Rollinia pittieri leaves was the most selective against Trypanosoma cruzi and Leishmania spp. (IS=10 and 16, respectively). The extracts from Desmopsis panamensis, Pseudomalmea boyacana, Rollinia exsucca and Rollinia pittieri showed good antiplasmodial activity (IC50 < 10 microg/ml). No correlation between antiplasmodial activity and inhibition of beta-hematin production was found. The present study gives specific and useful information about antiprotozoal and cytotoxic activities of some Annonaceae extracts. Results presented here also demonstrate which plants and/or plant parts could be useful in the treatment of leishmaniasis, Chagas' disease and malaria. PMID: 17296281 [PubMed - indexed for MEDLINE] Related articles

• _ Antiprotozoal and cytotoxic screening of 45 plant extracts from Democratic Republic of Congo. • J Ethnopharmacol. 2008 Feb 12; 115(3):409-15. Epub 2007 Dec 18. [J Ethnopharmacol. 2008] • _ Evaluation of antiprotozoal and plasmodial enoyl-ACP reductase inhibition potential of turkish

medicinal plants. Phytother Res. 2005 Feb; 19(2):162-6. [Phytother Res. 2005] • _ In vitro antiprotozoal activities and cytotoxicity of some selected Cameroonian medicinal plants. J

Ethnopharmacol. 2007 Apr 20; 111(1):8-12. Epub 2006 Nov 10. [J Ethnopharmacol. 2007] • _ A review of natural products with antileishmanial activity. Phytomedicine. 2005 Jun; 12(6-7):514-35.

[Phytomedicine. 2005] • _ Diamidines as antitrypanosomal, antileishmanial and antimalarial agents. Curr Opin Investig Drugs.

2006 Feb; 7(2):147-57. [Curr Opin Investig Drugs. 2006] 1: J Ethnobiol Ethnomed. 2006 Oct 13;2:45 Ethnomedicines used in Trinidad and Tobago for urinary problems and diabetes mellitus. Lans CA. BCICS, University of Victoria, British Columbia, V8W 2Y2, Canada. cher2lans@netscape.net BACKGROUND: This paper is based on ethnobotanical interviews conducted from 1996-2000 in Trinidad and Tobago with thirty male and female respondents. METHODS: A non-experimental validation was conducted on the plants used for urinary problems and diabetes mellitus: This is a preliminary step to establish that the plants used are safe or effective, to help direct clinical trials, and to inform Caribbean physicians of the plants' known properties to avoid counter-prescribing. RESULTS: The following plants are used to treat diabetes: Antigonon leptopus, Bidens alba, Bidens pilosa, Bixa orellana, Bontia daphnoides, Carica papaya, Catharanthus roseus, Cocos nucifera, Gomphrena globosa, Laportea aestuans, Momordica charantia, Morus alba, Phyllanthus urinaria and Spiranthes acaulis. Apium graviolens is used as a heart tonic and for low blood pressure. Bixa orellana, Bontia daphnoides, Cuscuta americana and Gomphrena globosa are used for jaundice. The following plants are used for hypertension: Aloe vera, Annona muricata, Artocarpus altilis, Bixa orellana, Bidens alba, Bidens pilosa, Bonta daphnoides, Carica papaya, Cecropia peltata, Citrus paradisi, Cola nitida, Crescentia cujete, Gomphrena globosa, Hibiscus sabdariffa, Kalanchoe pinnata, Morus alba, Nopalea cochinellifera, Ocimum campechianum, Passiflora quadrangularis, Persea americana and Tamarindus indicus. The plants used for kidney problems are Theobroma cacao, Chamaesyce hirta, Flemingia strobilifera, Peperomia rotundifolia, Petiveria alliacea, Nopalea cochinellifera, Apium graveolens, Cynodon dactylon, Eleusine indica, Gomphrena globosa, Pityrogramma calomelanos and Vetiveria zizanioides. Plants are also used for gall stones and for cooling. CONCLUSION: Chamaesyce hirta, Cissus verticillata, Kalanchoe pinnata, Peperomia spp., Portulaca oleraceae, Scoparia dulcis, and Zea mays have sufficient evidence to support their traditional use for urinary problems, "cooling" and high cholesterol. Eggplant extract as a hypocholesterolemic agent has some support but needs more study. The plants used for hypertension, jaundice and diabetes that may be safe and justify more formal evaluation are Annona squamosa, Aloe vera, Apium graveolens, Bidens alba, Carica papaya, Catharanthus roseus, Cecropia peltata, Citrus paradisi, Hibsicus sabdariffa, Momordica charantia, Morus alba,

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Persea americana, Phyllanthus urinaria, Tamarindus indicus and Tournefortia hirsutissima. Several of the plants are used for more than one condition and further trials should take this into account. 607BPMID: 17040567 [PubMed - indexed for MEDLINE] 608BPMCID: PMC1624823 609BRelated articles

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• 611B_ Comparison of plants used for skin and stomach problems in Trinidad and Tobago with Asian ethnomedicine. J Ethnobiol Ethnomed. 2007 Jan 5; 3:3. Epub 2007 Jan 5.

• 612B[J Ethnobiol Ethnomed. 2007] Prev Vet Med. 2000 Jun 12; 45(3-4):201-20. • 613B[Prev Vet Med. 2000] • 614B_ Medicinal and ethnoveterinary remedies of hunters in Trinidad. BMC Complement Altern Med. 2001;

1:10. Epub 2001 Nov 30. [BMC Complement Altern Med. 2001] • 615B_ Medicinal plants of India with anti-diabetic potential. J Ethnopharmacol. 2002 Jun; 81(1):81-100. [J

Ethnopharmacol. 2002] 616B1:J Neural Transm Suppl. 2006;(70):153-7. 617B_ Final report on the safety assessment of capsicum annuum extract, capsicum annuum fruit extract, capsicum annuum resin, capsicum annuum fruit powder, capsicum frutescens fruit, capsicum frutescens fruit extract, capsicum frutescens resin, and capsaicin. Int J Toxicol. 2007; 26 Suppl 1:3-106. [Int J Toxicol. 2007] 618B1:Nat Prod Res. 2006 Mar;20(3):253-7. 619BAcetogenins in Annona muricata L. (annonaceae) leaves are potent molluscicides. 620BLuna Jde S, De Carvalho JM, De Lima MR, Bieber LW, Bento Ede S, Franck X, Sant'ana AE. 621BDepartamento de Química Fundamental, Universidade Federal de Pernambuco, 50.740-901 Recife-PE, Brazil. 622BAn ethanolic extract of the leaves of Annona muricata was shown to be toxic to adult forms of the snail Biomphalaria glabrata (LC50 9.32 microg mL(-1)) and to larvae of the brine shrimp Artemia salina (LC50 0.49 microg mL(-1)). Activity-guided fractionation of the extract gave rise to a sample with high molluscicidal activity that contained the acetogenins, annonacin (90%), isoannonacin (6%) and goniothalamicin (4%). 623BPMID: 16401556 [PubMed - indexed for MEDLINE] 624B1:Nat Prod Res. 2006 Jan;20(1):21-6 625BAntibacterial activity of eight Brazilian annonaceae plants. 626BTakahashi JA, Pereira CR, Pimenta LP, Boaventura MA, Silva LG. 627BDepartamento de Química -- ICEx -- Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, CEP 31270-901, Belo Horizonte, MG, Brazil. 628Bjacfab@dedalus.lcc.ufmg.br 629BSixteen extracts, obtained from eight Brazilian plants of Annonaceae family, were screened for their antibacterial activity: Xylopia frutescens, X. aromatica, X. amazonica, X. benthamii, Annona ambotay, A. crassiflora, A. muricata and A. cherimolia. Amongst the investigated extracts, six showed antibacterial activity against at least one of the tested organisms at the concentration of 100 microg/mL. The most active extracts were those prepared from X. frutescens, X. amazonica, and A. ambotay. A phytochemical screening showed the presence of anonaceus acetogenins in some active extracts. Eleven diterpenoids were also tested for comparison purposes. Six were natural products, previously isolated from Xylopia sp. (kaurenoic, frutoic, xylopic, 15beta-hydroxy-kaurenoic and trachylobanic acids plus kaurenol) and five were derivatives of such compounds, obtained by esterification or reduction reactions. Trachylobanic acid showed antibacterial activity against B. subtilis and S. aureus. 630BPMID: 16286303 [PubMed - indexed for MEDLINE]

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Prod Rep. 2005 Apr; 22(2):269-303. Epub 2005 Mar 15. [Nat Prod Rep. 2005] • _ [Chemical constituents of annonaceae plants and their antitumor activities] Zhongguo Yi Xue Ke Xue

Yuan Xue Bao. 2000 Aug; 22(4):376-82. [Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2000] 1:Mov Disord. 2005 Dec;20(12):1629-33

1:Int J Food Sci Nutr. 2004 Aug;55(5):407-14 Nutritional and sensory quality of stirred soursop (Annona muricata L.) yoghurt. Lutchmedial M, Ramlal R, Badrie N, Chang-Yen I. Department of Chemistry, Faculty of Science and Agriculture, University of the West Indies, St Augustine, Republic of Trinidad and Tobago, West Indies. Soursops (Annona muricata L.) are highly aromatic fruits with white juicy flesh and are native to tropical North and South America. The ripe fruits are highly perishable, as they become soft and easily bruised. The objectives of the study were to incorporate soursop nectar at 0%, 5%, 10% and 15% in stirred yoghurts and to analyse the products for chemical and sensory quality. A focus group evaluated the initial yoghurts for process modifications. Yoghurts were evaluated on sensory attributes of appearance and colour, body and texture, flavour and aroma, and overall quality. Yoghurts with 10% and 15% soursop nectar had the highest (P<0.05) overall quality scores (12.60/20 and 12.75/20, respectively) but differed (P<0.05) in flavour and aroma from plain yoghurt and 5% soursop yoghurt. Most panelists would consider purchase of 10% and 15% soursop yoghurts over 0% and 5% soursop yoghurts. These yoghurts provided high percentage daily values of zinc, phosphorus and calcium and a good level of protein. PMID: 15545049 [PubMed - indexed for MEDLINE] Related articles

• _ Physicochemical, nutritional and sensory quality of stirred 'dwarf' golden • apple (Spondias cytherea Sonn) yoghurts. Int J Food Sci Nutr. 2005 Sep; 56(6):445-54. • [Int J Food Sci Nutr. 2005] • _ Development and sensory evaluation of soy milk based yoghurt. Arch Latinoam Nutr. 2001 Mar;

51(1):100-4. [Arch Latinoam Nutr. 2001] • _ [A transcultural study on stirred strawberry yogurt: consumer acceptability versus sensory quality with

trained panel] Arch Latinoam Nutr. 2005 Mar; 55(1):77-85. [Arch Latinoam Nutr. 2005] • _ The impact of processing on the nutritional quality of food proteins. J AOAC Int. 2005 May-Jun;

88(3):904-22. [J AOAC Int. 2005] • _ Safety and nutritional assessment of GM plants and derived food and feed: the role of animal feeding

trials. Food Chem Toxicol. 2008 Mar; 46 Suppl 1:S2-70. Epub 2008 Feb 13. [Food Chem Toxicol. 2008]

1:Tree Physiol. 2004 Sep;24(9):1019-25 Flooding, root temperature, physiology and growth of two Annona species. Ojeda M, Schaffer B, Davies FS. Horticultural Sciences Department, University of Florida, IFAS, P.O. Box 110690, Gainesville, FL 32611, USA. The effects of root zone temperature (RZT) and flooding on physiology and growth of Annona glabra L. (pond apple) and A. muricata L. (soursop) were investigated. Trees planted in containers were exposed to RZTs of 5,

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10, 20, 25 or 35 degrees C in controlled root temperature chambers. Trees at each RZT were either non-flooded (control) or continuously flooded. There were four replications over time for each treatment combination. Pond apple was more flood-tolerant than soursop. A combination of flooding and RZTs of 5 and 10 degrees C resulted in tree mortality of both species by Week 4. Only trees that appeared to develop morphological adaptations survived continuous flooding. In both species, net CO2 assimilation (A) decreased to nearly zero within 1 week following exposure to RZTs of 5 or 10 degrees C and became consistently negative over the remaining experimental period. Flooding reduced leaf chlorophyll index (measured with a SPAD meter), A and plant growth, and increased root electrolyte leakage from soursop. Optimum growth occurred at RZTs of 25 to 35 degrees C for non-flooded pond apple trees and at 20 to 25 degrees C for flooded trees. Soursop exhibited maximum growth at RZTs of 35 degrees C under non-flooded conditions and at 25 degrees C under flooded conditions. 656BPMID: 15234899 [PubMed - indexed for MEDLINE] 657B1:J Chromatogr B Analyt Technbol Biomed Life Sci 2004 Jun 658B25;806(1):75 659BDetection and determination of reticuline and N-methylcoculaurine in the Annonaceae family using liquid chromatography-tandem mass spectrometry. 660BKotake Y, Okuda K, Kamizono M, Matsumoto N, Tanahashi T, Hara H, Caparros-Lefebvre D, Ohta S. 661BGraduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8551, Japan. yaichiro@hiroshima-u.ac.jp In Guadeloupe, the French West Indies, there is a high incidence of atypical parkinsonism or progressive supranuclear palsy, and all of the investigated patients had taken herbal tea or tropical fruits of the Annonaceae family. Local inhabitants consume the fruits, and also drink tea made from the leaves. In the present study, we used liquid chromatography-tandem mass spectrometry (LC/MS/MS) with multiple reaction monitoring (MRM) to detect lowmolecular- weight neurotoxic benzylisoquinoline derivatives in the Annonaceae family. We detected reticuline and N-methylcoculaurine in every Annona muricata sample examined, except for pulp and seed. They were not detected in sweetsop fruits. Norreticuline was not detected in any sample. These three compounds were toxic to SH-SY5Y neuroblastoma cells and inhibited mitochondrial respiratory complex I. It is possible that uptake of the benzylisoquinoline derivatives reticuline and N-methylcoculaurine and their accumulation in the brain may be related to the pathogenesis of the local endemic disease. 662BPMID: 15149614 [PubMed - indexed for MEDLINE] 663BRelated articles

• 664B_ Possible relation of atypical parkinsonism in the French West Indies with consumption of tropical plants: a case-control study. Caribbean Parkinsonism Study Group. Lancet. 1999 Jul 24; 354(9175):281-6. [Lancet. 1999]

• 665B_ Analysis of benzylisoquinoline-type alkaloids by electrospray tandem mass spectrometry and atmospheric pressure photoionization. Eur J Mass Spectrom (Chichester, Eng). 2005; 11(3):325-33. [Eur J Mass Spectrom (Chichester, Eng). 2005]

• 666B_ Toxicity of Annonaceae for dopaminergic neurons: potential role in atypical parkinsonism in Guadeloupe. Mov Disord. 2002 Jan; 17(1):84-90. [Mov Disord. 2002] J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Jul 1; 854(1-2):1- 7. Epub 2007 Apr 5. [J Chromatogr B Analyt Technol Biomed Life Sci. 2007] Anal Bioanal Chem. 2005 Jun; 382(4):934-46. Epub 2005 May 25. [Anal Bioanal Chem. 2005]

667B1:J Protein Chem 2003 Nov;22(7-8):655-61. 668BIsolation and characterization of a lectin from Annona muricata seeds. 669BDamico DC, Freire MG, Gomes VM, Toyama MH, Marangoni S, Novello JC, Macedo ML. 670BDepartamento de Bioquímica, Instituto de Biología, Universidade Estadual de Campinas, Campinas (SP), Brazil.

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A lectin with a high affinity for glucose/mannose was isolated from Annona muricata seeds (Annonaceae) by gel filtration chromatography on Sephacryl S- 200, ion exchange chromatography on a DEAE SP-5 PW column, and molecular exclusion on a Protein Pak Glass 300 SW column. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (PAGE) yielded two protein bands of approximately 14 kDa and 22 kDa. However, only one band was seen in native PAGE. The Mr of the lectin estimated by fast-performance liquid chromatography-gel filtration on Superdex 75 was 22 kDa. The lectin was a glycoprotein with 8% carbohydrate (neutral sugar) and required divalent metal cations (Ca2+, Mg2+, and Mn2+) for full activity. Amino acid analysis revealed a large content of Glx, Gly, Phe, and Lys. The lectin agglutinated dog, chicken, horse, goose, and human erythrocytes and inhibited the growth of the fungi Fusarium oxysporum, Fusarium solani, and Colletotrichum musae. PMID: 14714732 [PubMed - indexed for MEDLINE] Related articles

• _ Isolation and partial characterization of a lectin from Artocarpus incisa L. • seeds. Phytochemistry. 1998 Apr; 47(7):1183-8. [Phytochemistry. 1998] • _ Purification, some properties of a D-galactose-binding leaf lectin from Erythrina indica and further

characterization of seed lectin. Biochimie. 2002 Oct; 84(10):1035-43. • [Biochimie. 2002] • _ Purification and characterization of a lectin from wild sunflower (Helianthus tuberosus L.) tubers.

Arch Biochem Biophys. 2002 Nov 15; 407(2):241-7. [Arch Biochem Biophys. 2002] • _ [Isolation of lectin from horse chestnut (Aesculus hippocastanum L.) seeds and study of its interaction

with carbohydrates and glycoproteins] Ukr Biokhim Zh. 1992 Sep-Oct; 64(5):47-52. [Ukr Biokhim Zh. 1992]

• _ Purification and characterization of a lectin from Annona coriacea seeds. Protein Pept Lett. 2003 Apr; 10(2):165-73. [Protein Pept Lett. 2003]

1:J Neurochem. 2004 Jan;88(1):63-9 1:J Pharm Biomed Anal. 2003 Dec 4;33(5):1025-31. Flow-injection amperometric determination of dopamine in pharmaceuticals using a polyphenol oxidase biosensor obtained from soursop pulp. Bezerra VS, de Lima Filho JL, Montenegro MC, Araújo AN, da Silva VL. Laboratory of Immunopathology Keizo Asami (LIKA), Department of Biochemistry, UFPE, 50670-901 Recife PE, Brazil. The amperometric determination of dopamine (Do) in pharmaceuticals formulations by flow injection analysis (FIA) is proposed. An enzymatically modified carbon paste electrode constituted by 25% (w/w) of polyphenol oxidase obtained from Annona muricata L. tissue, 30% (w/w) of graphite, 30% (w/w) of silicone and 15% (w/w) of 7,7,8,8 tetracyanoquinodimethane (TCNQ), was used as flow-through detector. The flow amperometric detection was carried out at a potential of 0.10 V (vs. Ag/AgCl) when an injected sample volume of 250 microl was inserted on a 0.3 M phosphate buffer carrier solution (pH 7.8) flowing at 2.5 ml/min. The developed biosensor showed good stability and reproducibility, enabling up to 500 determinations in 60 days, without considerable loss of enzymatic activity. The FIA system presented a linear response to Do concentrations in the interval from 2 x 10(-2) to 2 x 10(-4) M, with relative standard deviations lower than 1.5%. The kinetic parameter K(M) for the soluble and immobilized enzyme was 1.45 x 10(-2) and 1.91 x 10(-2) M, respectively. In the analyses of different commercially pharmaceutical formulations a relative deviation lower than about 3.4% was obtained. PMID: 14656593 [PubMed - indexed for MEDLINE] Related articles

• _ Flow injection amperometric determination of dipyrone in pharmaceutical formulations using a carbon paste electrode. Farmaco. 2003 Oct; 58(10):999-1004. [Farmaco. 2003]

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• 690B_ An amperometric flow-injection analysis biosensor for glucose based on graphite paste modified with tetracyanoquinodimethane. Anal Biochem. 1993 Oct; 214(1):233-7. [Anal Biochem. 1993]

• 691B_ Flow injection analysis with amperometric detection of naltrexone in pharmaceuticals. J Pharm Biomed Anal. 1997 Sep; 16(1):15-9. [J Pharm Biomed Anal. 1997]

692B1:Neuroscience. 2003;121(2):287-96. 693B1:Planta Med 2003 Mar;69(3):241-6. 694BNew adjacent Bis-tetrahydrofuran Annonaceous acetogenins from Annona muricata. 695BChang FR, Liaw CC, Lin CY, Chou CJ, Chiu HF, Wu YC. 696BGraduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC. Bioactivity-guided fractionation led to the isolation of two new Annonaceous acetogenins, annocatacin A ( 1). and annocatacin B ( 2). from the seeds and the leaves, respectively, of Annona muricata. Compounds 1 and 2 are the first examples where the adjacent bis-tetrahydrofuran ring system is located at C-15. The new structures were elucidated and characterized by spectral and chemical methods. Both Annonaceous acetogenins 1 and 2 showed significant in vitro cytotoxicity toward the human hepatoma cell lines, Hep G2 and 2,2,15, and were compared with the known adjacent bis-tetrahydrofuran acetogenins, neoannonin ( 3). desacetyluvaricin ( 4). bullatacin ( 5). asimicin ( 6). annoglaucin ( 7). squamocin ( 8). and rollimusin ( 9). 697BPMID: 12677528 [PubMed - indexed for MEDLINE] 698BRelated articles

• 699B_ Novel cytotoxic annonaceous acetogenins from Annona muricata. J Nat Prod. 2001 Jul; 64(7):925-31. [J Nat Prod. 2001]

• 700B_ Purpurediolin and purpurenin, two new cytotoxic adjacent bistetrahydrofuran annonaceous acetogenins from the seeds of Annona purpurea. J Nat Prod. 1998 May; 61(5):580-4. [J Nat Prod. 1998]

• 701B_ Nine new cytotoxic monotetrahydrofuranic Annonaceous acetogenins from Annona montana. Planta Med. 2004 Oct; 70(10):948-59. [Planta Med. 2004]

• 702B_ Recent advances in Annonaceous acetogenins. Nat Prod Rep. 1996 Aug; 13(4):275-306. [Nat Prod Rep. 1996]

• 703B_ Annonaceous acetogenins: recent progress. J Nat Prod. 1999 Mar; 62(3):504-40. [J Nat Prod. 1999] 704B1:Plant Foods Hum Nutr 2002 Spring;57(2):165-71. 705BProximate composition and selected physicochemical properties of the seed, 706Bpulp and oil of sour sop (Annona muricata). 707BOnimawo IA. 708BBiochemistry Department, Ambrose Alli University, Edo State, Nigeria. Proximate composition and physicochemical analyses were carried out on the seed, pulp and extracted oil of sour sop (Annona muricata). The results showed that the seed contained 8.5% moisture, 2.4% crude protein, 13.6% ash, 8.0% crude fiber, 20.5% fat and 47.0% carbohydrate. The seed also contained 0.2% water soluble ash, 0.79% titratable acidity and 17.0 mg calcium/100 g. The pulp was found to contain 81% moisture, 3.43% titratable acidity and 24.5% nonreducing sugar. Selected physicochemical characteristics included refractive indices of 1.335 for the seed and 1.356 for the pulp, specific gravities of 1.250 for the seed and 1.023 for the pulp, pH values of 8.34 for the seed and 4.56 for the pulp, and soluble solids contents of 1.5 degrees Brix for the seed and 15 degrees Brix for the pulp. The extracted oil (20.5% yield) had a 60.43% unsaponifiable value, 23.54 KOH/g acid value, 100.98 KOH/g saponification value, 1.1 KOH/g peroxide value, 1.464 refractive index, 5.77 pH, 69.5 degrees Brix sosluble solids and 0.2900 specific gravity. 709BPMID: 12049148 [PubMed - indexed for MEDLINE] 710BRelated articles 711B_ Chemical composition of seeds and oil of Xylopia aethiopica grown in Nigeria. 712BPlant Foods Hum Nutr. 1999; 53(3):193-8. [Plant Foods Hum Nutr. 1999]

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_ Proximate composition and selected functional properties of Detarium microcarpum. Plant Foods Hum Nutr. 2001; 56(4):297-302. [Plant Foods Hum Nutr. 2001] _ [Chemical characteristics of the pulp and oil of the annona tree (Annona coriaceae)] Arch Latinoam Nutr. 1995 Sep; 45(3):237-41. [Arch Latinoam Nutr. 1995] _ Baobab food products: a review on their composition and nutritional value. Crit Rev Food Sci Nutr. 2009 Mar; 49(3):254-74. [Crit Rev Food Sci Nutr. 2009] _ Nutritional and functional properties of dates: a review. Crit Rev Food Sci Nutr. 2008 Nov; 48(10):877-87. [Crit Rev Food Sci Nutr. 2008] 1:J Nat Prod. 2002 Apr;65(4):470-5. New cytotoxic monotetrahydrofuran annonaceous acetogenins from Annona muricata. Liaw CC, Chang FR, Lin CY, Chou CJ, Chiu HF, Wu MJ, Wu YC. Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung 807, Taiwan, Republic of China. Three new monotetrahydrofuran annonaceous acetogenins, muricin H (1), muricin I (2), and cis-annomontacin (3), along with five known acetogenins, annonacin, annonacinone, annomontacin, murisolin, and xylomaticin, were isolated from the seeds of Annona muricata. Additionally, two new monotetrahydrofuran annonaceous acetogenins, cis-corossolone (4) and annocatalin (5), together with four known ones, annonacin, annonacinone, solamin, and corossolone, were isolated from the leaves of this species. The structures of all new isolates were elucidated and characterized by spectral and chemical methods. These new acetogenins exhibited significant activity in in vitro cytotoxic assays against two human hepatoma cell lines, Hep G(2) and 2,2,15. Compound 5 showed a high selectivity toward the Hep 2,2,15 cell line. PMID: 11975482 [PubMed - indexed for MEDLINE] Related articles

• _ Montacin and cis-montacin, two new cytotoxic monotetrahydrofuran annonaceous acetogenins from Annona montana. J Nat Prod. 2004 Nov; 67(11):1804-8. [J Nat Prod. 2004]

• _ Novel cytotoxic annonaceous acetogenins from Annona muricata. J Nat Prod. 2001 Jul; 64(7):925-31. [J Nat Prod. 2001]

• _ Muricatocins A and B, two new bioactive monotetrahydrofuran Annonaceous acetogenins from the leaves of Annona muricata. J Nat Prod. 1995 Jun; 58(6):902-8. [J Nat Prod. 1995]

• _ Annonaceous acetogenins: recent progress. J Nat Prod. 1999 Mar; 62(3):504-40. [J Nat Prod. 1999] • _ Acetogenins from Annonaceae. Fortschr Chem Org Naturst. 1997; 70:81-288. [Fortschr Chem Org

Naturst. 1997] 1:Mov Disord. 2002 Jan;17(1):84-90. Cytotoxicity and antileishmanial activity of Annona muricata pericarp - graviola. Fitoterapia. 2000 Apr;71(2):183-6. Hexane, ethyl acetate and methanol extracts of Annona muricata pericarp ( graviola ) were tested in vitro against Leishmania braziliensis and L. panamensis promastigotes, and against cell line U-937. The ethyl acetate graviola extract was more active than the other extracts and even of Glucantime used as reference substance. Its fractionation led to the isolation of three acetogenins--annonacin, annonacin A and annomuricin A. Two new mono-tetrahydrofuran ring acetogenins, annomuricin E and muricapentocin, from the leaves of Annona muricata - graviola. J Nat Prod. 1998 Apr;61(4):432-6. Bioactivity-directed fractionation of the leaf extract of Annona muricata L. (graviola) has resulted in the isolation of two new Annonaceous acetogenins, annomuricine (1) and muricapentocin (2). Compounds 1 and 2 are monotetrahydrofuran ring acetogenins bearing two flanking hydroxyl groups; however, each has three additional hydroxyl groups. Compound 1 has an erythro 1,2-diol, and 2 has a 1,5,9-triol moiety. Both 1 and 2 showed significant cytotoxicities against six types of human tumors, with selectivities to the pancreatic carcinoma (PACA-2) and colon adenocarcinoma (HT-29) cell lines. Graviola research.

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733BIsoquinoline derivatives isolated from the fruit of Annona muricata ( graviola ) as 5- HTergic 5-HT1A receptor agonists in rats: unexploited antidepressive (lead) products. J Pharm Pharmacol. 1997 Nov;49(11):1145-9. 734BExtracts of the graviola plant have been shown to inhibit binding of [3H]rauwolscine to 5-HTergic 5-HT1A receptors in calf hippocampus, and three alkaloids, annonaine (1), nornuciferine (2) and asimilobine (3), isolated from the fruit have been shown to have IC50 values of 3 microM, 9 microM and 5 microM, respectively, although in ligandbinding studies it was not possible to determine whether interaction of these ligands with the receptor was agonistic or antagonistic. These results imply that the fruit of graviola possesses anti-depressive effects, possibly induced by compounds 1, 2 and 3, and that in the past potent leads for the development of anti-depressive therapeutics have not been used. 735BFive new monotetrahydrofuran ring acetogenins from the leaves of Annona 736Bmuricata - graviola. 737BJ Nat Prod. 1996 Nov;59(11):1035-42. 738BBioactivity-directed fractionation of the leaves of Annona muricata ( graviola ) resulted in the isolation of annopentocins A (1), B (2), and C(3), and cis- and trans-annomuricin- D-ones (4, 5). Compounds 1-3 are the first acetogenins reported bearing a monotetrahydrofuran (THF) ring with one flanking hydroxyl, on the hydrocarbon side, and another hydroxyl, on the lactone side, that is one carbon away from the THF ring. Compounds 4 and 5 from graviola were obtained in a mixture and are new mono-THF ring acetogenins bearing two flanking hydroxyls and an erythro-diol located between the THF and the ketolactone rings. Compound 1 was selectively cytotoxic to pancreatic carcinoma cells (PACA-2), and 2 and 3 were selectively cytotoxic to lung carcinoma cells (A-549); the mixture of 4 and 5 was selectively cytotoxic for the lung (A-549), colon (HT-29), and pancreatic (PACA-2) cell lines with potencies equal to or exceeding those of Adriamycin. Graviola anti-cancer benefits. 739BFive novel mono-tetrahydrofuran ring acetogenins from the seeds of Annona muricata ( graviola ). J Nat Prod. 1996 Feb;59(2):100-8. 740BBioactivity-directed fractionation of the seeds of Annona muricata L. ( graviola ) resulted in the isolation of five new compounds: cis-annonacin (1), cis-annonacin-10-one (2), cisgoniothalamicin (3), arianacin (4), and javoricin (5). Three of these (1-3) are among the first cis mono-tetrahydrofuran ring acetogenins to be reported. NMR analyses of published model synthetic compounds, prepared cyclized formal acetals, and prepared Mosher ester derivatives permitted the determinations of absolute stereochemistries. Bioassays of the pure graviola compounds, in the brine shrimp test, for the inhibition of crown gall tumors, and in a panel of human solid tumor cell lines for cytotoxicity, evaluated relative potencies. Compound 1 from graviola was selectively cytotoxic to colon adenocarcinoma cells (HT-29) in which it was 10,000 times the potency of adriamycin. 741BMuricatocins A and B, two new bioactive monotetrahydrofuran Annonaceous acetogenins from the leaves of Annona muricata (graviola). 742BJ Nat Prod. 1995 Jun;58(6):902-8. 743BThe leaves of Annona muricata (graviola) have yielded the novel monotetrahydrofuran Annonaceous acetogenins, muricatocins A [1] and B [2]. Each compound possesses five hydroxyl groups, with two hydroxyl groups at the C-10 and C-12 positions. The absolute configurations of 1 and 2 (except for positions C-10 and C-12) were determined by Mosher ester methodology. The C-10, C-12 acetonides (1c, 2c) suggested relative stereochemistry and significantly enhanced cytotoxicity against the A-549 human lung tumor cell line. Three known monotetrahydrofuran acetogenins, annonacin A, (2,4-trans)- isoannonacin, and (2,4-cis)-isoannonacin, were also found from graviola.

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Two new cytotoxic monotetrahydrofuran Annonaceous acetogenins, annomuricins A and B, from the leaves of Annona muricata ( graviola ). J Nat Prod. 1995 Jun;58(6):830-6. The leaves of graviola have yielded eight monotetrahydrofuran Annonaceous acetogenins. Two of them, annomuricins A [1] and B [2], whose chemical structures were deduced by ms, nmr, ir, and uv spectral and chemical methods, are novel and unusual. Compounds 1 and 2 each possess five hydroxyl groups; two hydroxyl groups are vicinal, with the vicinal group of 1 threo and that of 2 erythro. The absolute configurations of 1 and 2 were determined by Mosher ester methodology. Six monotetrahydrofuran acetogenins, previously described in the graviola seeds, were found in the graviola leaves; these are gigantetrocin A, annonacin-10-one, muricatetrocins A and B, annonacin, Antibacterial activity of eight Brazilian annonaceae plants. Takahashi JA, Pereira CR, Pimenta LP, Boaventura MA, Silva LG. Departamento de Química -- ICEx -- Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, CEP 31270-901, Brazil Sixteen extracts, obtained from eight Brazilian plants of Annonaceae family, were screened for their antibacterial activity: Xylopia frutescens, X. aromatica, X. amazonica, X. benthamii, Annona ambotay, A. crassiflora, A. muricata and A. cherimolia. Amongst the investigated extracts, six showed antibacterial activity against at least one of the tested organisms at the concentration of 100 microg/mL. The most active extracts were those prepared from X. frutescens, X. amazonica, and A. ambotay. A phytochemical screening showed the presence of anonaceus acetogenins in some active extracts. Eleven diterpenoids were also tested for comparison purposes. Six were natural products, previously isolated from Xylopia sp. (kaurenoic, frutoic, xylopic, 15beta-hydroxy-kaurenoic and trachylobanic acids plus kaurenol) and five were derivatives of such compounds, obtained by esterification or reduction reactions. Trachylobanic acid showed antibacterial activity against B. subtilis and S. aureus. PMID: 16286303 [PubMed - indexed for MEDLINE] Related articles

• _ Antimicrobial activity of some species of the family Combretaceae. Phytother Res. 2002 Sep; 16(6):555-61. [Phytother Res. 2002]

• _ HPLC quantitation of kaurane diterpenes in Xylopia species. Fitoterapia. 2001 Jan; 72(1):40-5. [Fitoterapia. 2001]

• _ Antibacterial activity of plant extracts from northwestern Argentina. J Appl Microbiol. 2007 Jun; 102(6):1450-61. [J Appl Microbiol. 2007]

• _ Acetogenins from Annonaceae: recent progress in isolation, synthesis and mechanisms of action. Nat Prod Rep. 2005 Apr; 22(2):269-303. Epub 2005 Mar 15. [Nat Prod Rep. 2005]

• _ [Chemical constituents of annonaceae plants and their antitumor activities] Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2000 Aug; 22(4):376-82. [Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2000]

In vitro antioxidant studies in leaves of Annona species. Baskar R, Rajeswari V, Kumar TS. Department of Biotechnology, Kumaraguru College of Technology, India. Antioxidant potential of leaves of three different species of Annona was studied by using different in vitro models eg., 1,1-diphenyl-2-picryl hydrazyl (DPPH), 2,2-azinobis-(3-ethylbenzothizoline-6-sulphonate) (ABTS), nitric oxide, superoxide, hydroxy radical and lipid peroxidation. The ethanolic extract of A. muricata at 500 microg/ml showed maximum scavenging activity (90.05%) of ABTS radical cation followed by the scavenging of hydroxyl radical (85.88%) and nitric oxide (72.60%) at the same concentration. However, the extract showed only moderate lipid peroxidation inhibition activity. In contrast, the extract of A. reticulata showed better activity in quenching DPPH (89.37%) and superoxide radical (80.88%) respectively. A.squamosa extract exhibited least inhibition in all in vitro antioxidant models excepting hydroxyl radical (79.79%). These findings suggest that the extracts of A. muricata possess potent in vitro antioxidant activity as compared to leaves of A. squamosa and A. reticulata suggesting its role as an effective free radical scavenger, augmenting its therapeutic PMID: 17569293 [PubMed - indexed for MEDLINE]

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760BRelated articles • 761B_ In vitro antioxidant studies of Annona squamosa Linn. leaves. Indian J Exp Biol. 2004 Aug;

42(8):803-7. [Indian J Exp Biol. 2004] • 762B_ In vitro antioxidant studies of Sphaeranthus indicus (Linn). Indian J Exp Biol. 2006 Dec; 44(12):993-

6. [Indian J Exp Biol. 2006] • 763B_ In vitro antioxidant activity of Diospyros malabarica Kostel bark. Indian J Exp Biol. 2006 Jan;

44(1):39-44. [Indian J Exp Biol. 2006] • 764B_ Cytisus scoparius link--a natural antioxidant. BMC Complement Altern Med. 2006 Mar 16; 6:8. Epub

2006 Mar 16. [BMC Complement Altern Med. 2006] • 765B_ Antioxidant activity of Caesalpinia digyna root. J Ethnopharmacol. 2007 Sep 5; 113(2):284-91. Epub

2007 Jun 26. [J Ethnopharmacol. 2007]