TW200407119A - Methods and compositions to treat conditions associated with neovascularization - Google Patents

Abstract

This invention provides a method to inhibit neovascularization in tissue by delivering to the cell or tissue an effective amount of sulforaphane, or a pharmaceutically acceptable salt, derivative or prodrug thereof. Also provided herein is a method for treating a disease associated with hyperproliferation of endothelial cells and/or neovascularization by administering to a subject an effective amount of sulforaphane, or a pharmaceutically acceptable salt, derivative or prodrug thereof. Kits to treat patients are provided as well.

Description

200407119 (1) 发明, related application with cross-reference to invention description This application is a US provisional application No. 6 0/4 0 1 J 3 0 which is claimed as 35 US C & 119 (e), application date 2 The rights and interests of August 5, 2002, the contents of which are hereby incorporated herein by reference. [Technical Field to which the Invention belongs] The present invention belongs to the field of medicine. In particular, they are related to antiangiogenic drugs to prevent and treat diseases. [Prior art] Angiogenesis is the creation of new vascular structures through the growth of existing microvessels. In this role, endothelial cells are released from the basement membrane by the degradation of proteolytic enzymes. Such cells then migrate, divide, and form newly differentiated vascular structures from the parent blood vessel (Risau, (1 997) Nature 3 8 6:67 1 -6 7 4; Wilting et al .. (1 99 5) Cell. Mol Biol. Res. 4 1 (4): 2 1 9-23 2). It has been found that the functions of different biological factors in the book can control angiogenesis (Bussolino et al., (1997) Trends in Biochem sci 22 (7): 251-256; Folkman and D'Amore, (1996), Cell 8 7: 1 1 5 3-1 1 5 5). Such factors include proteins with various functions, such as: growth factors, cell surface receptors, proteins, protein cells, protein inhibitors, and extracellular matrix proteins (A chenand Stacker, (198) Int. J. Exp. Pathol. 79: 255-265; Devalaraja and Richmond. (1999) Trends in Pharmacol. Sci. 20 (4): 1 51-156; Ha n ah an, (2) (2) 200407119 (1 99 7) Science 2 7 7: 4 8 -50; M aisonpierre et a]; (1 9 9 7) Science 2 7 7: 5 5-6 0; Suri et al5 (1 9 9 6) Cell 8 7: 1 1 7 1-1 1 8 0; Sato et a]. (1 9 9 5) Nature 376: 70-74; M igna 11 i and Rifkin, (1 9 9 6) Enzyme Protein 4 9: 1 1 7-1 3 7 ; P intuccie 1 a t.? (1 9 96) Semin thromb He most 22 (6) 517-524; Vernon and Sage, (1 995) Am. J. Pathol. 1 4 7 (4): 8 7 3- 8 8 3; Brooks et al. 5 (1 9 9) Science 264: 569-571; Koch et al. (1995)

Nature 3 7 6: 5 1 7-5 1 9). The complexity of angiogenic effects and the diversity of factors that control their progression provide many suitable therapeutic intervention points to control the formation of blood vessels in vivo. Angiogenesis usually occurs during embryogenesis, growth, and special circumstances, such as wound healing and the female reproductive cycle, and needs to be carefully controlled (W i 11 ing and Crist, (19 9 6) Naturewissenschaften 8 3: 1 5 3-1 64; Goodger and Rogers, (1 9 9 5) Microcirulation 2: 329-343; Augustin et al. 5 (1995) Am. J. Pathol. 14 7 (2): 339-35 1). Some important steps in angiogenesis are: 1) signals from growth factors (ie, vascular endothelial growth factor, VEGF); 2) matrix metalloproteinase (MMP) and VEGF receptor interactions: 3) endothelial cells migrate to The growth factor signal is the site: and 4) the formation of endothelial cell tubules. Pathological angiogenesis plays a major role in many human diseases, including tumor growth and migration cancer, diabetic retinopathy, rheumatoid arthritis, and other inflammatory diseases, such as: psoriasis (F 〇] kma η; (1 9 9 5) Nature Med.] (1): 2 7-3 1; P o 1 verini? (1 9 9 5) (3) (3) 200407119

Rheumatology 38 (2): 103-1 12; Healy et al.? (1998) Hum. Reprod. Update 4 (5): 7 3 6-3 9 6) In such cases, disease progression is unregulated and sustained Driven by sexual angiogenesis. For example, in rheumatoid arthritis, new microvessels invade joints and destroy cartilage. In diabetic retinopathy, microvessels of the retina invade the vitreous, causing bleeding and causing blindness. Importantly, tumor growth and metastasis are strongly related to angiogenesis. Most solid tumors are exposed to a long period of angiogenesis, during which time their growth is limited to 1-2 mm in diameter. At this size, tumor cells can obtain the necessary oxygen and nutrients through passive diffusion. Such tiny tumor masses will eventually start angiogenesis and recruit surrounding blood vessels, and allow the growing microvessels to supply blood to the tumor mass, providing the potential for continued tumor expansion and the migration of malignant cells to a distance. Although much progress has been made in understanding the elimination phenomenon that occurs during pathological angiogenesis, effective pharmaceutical compounds have not yet been developed to control angiogenesis in vivo. Therefore, effective treatments that control angiogenesis have soothing potential for many human diseases. Traditionally, investigating pharmaceutical compounds has been to screen chemical compounds with satisfactory medicinal properties and then test their toxicity and efficacy in vivo. The compounds selected by the sequential method often have in vivo toxic side effects, and this method has not been successful in suppressing the effective angiogenesis effect of the disease. Recently, molecular biology techniques have been used to develop inhibitors of hemorrhage S production. For example: angi statin (O'Reilly et al "(1 994) Cell 79 (2): 3 1 5 -3 2 8) and endostati η (Ο 5 R ei] 1 y et al., (1997) Cell 88 (2): 2 7 7- 285), (4) 200407119 Control the formation of blood vessels. However, the protein therapeutics are expensive to produce and found to be difficult to cultivate and deliver to each other. At present, 'protein angiogenesis inhibitors have not developed into therapeutic drugs. Therefore, people need a safe A therapeutic compound administered to a patient and effective in inhibiting pathological growth of vascular endothelial cells. The present invention provides a composition method suitable for this purpose and having related advantages.

[Summary of the Invention]

It has been found that calyxane and its derivatives can inhibit endothelial cell growth and proliferation and angiogenesis. Therefore, the present invention provides a method for inhibiting the proliferation of endothelial cells, especially for reaching specific pathological degree or specific cells in a tissue. The invention also provides a method for inhibiting the formation of new blood vessels in a tissue. Each method requires the delivery of an effective amount of rapamidine, or a pharmaceutically acceptable salt, derivative or prodrug thereof, to the cell or tissue. One method can be used in combination with other compounds, compositions, or other cells that inhibit the growth of new blood vessels or transitions, such as those that occur during cancer treatment. These treatments include, but are not limited to, chemotherapy, radiotherapy and the application of anti-neovascular compounds. The present invention also provides a method for treating diseases related to endothelial cell proliferation and / or neovascularization, which comprises administering to a patient an effective amount of calyxane, or a pharmaceutically acceptable salt, derivative or Prodrug. One method This invention can be combined with other compounds, composition 5 or with inhibition of neovascularization and transitional cell growth, such as in the treatment of cancer, these treatments include, but are not limited to, chemotherapy, radiotherapy and Suitable for anti-angiogenesis and compounds. The invention also provides a kit for treating patients. (5) (5) 200407119 The present invention further provides a method for screening new treatments to confirm that the effect of the oral treatment is the same as or similar to that of the Sulphur Treatment Center or its pharmaceutically acceptable salts, derivatives or prodrug Or better. Screening needs to compare the anti-hypertensive effects of glucosinolates, or their medically acceptable salts, derivatives, or prodrugs. The present invention further administers a compound, a composition, or a method and method for treating and preventing neovascular growth, angiogenesis, neoplasia, or cancer. These treatments include, but are not limited to, chemotherapy, radiotherapy, and the use of anti-neovascular growth compounds. [Embodiment] In this disclosure, various documents, patents, and published patent specifications are cited as references. The disclosures of such documents, patents, and published patent specifications are incorporated herein by reference in their entirety in order to more fully describe the state of the art of the present invention in this disclosure. Unless otherwise specified in the present invention, molecular biology (including the art of resection), microbiology, cell biology, biochemistry, and immune technology are all well-known techniques. This technique is fully explained in the literature. Definitions The meaning of certain terms in this document is defined as follows. The singular forms "a", "an", and "the" used in the description and the scope of the patent application include the meaning of the majority unless expressly stated otherwise. For example, "a cell" herein includes a majority of cells (6) 200407119 and includes mixtures thereof. The "package" in this document refers to the components and components included in the method, and does not send other components. When used to define composition, "essentially consisting of" shall exclude other important elements of the composition. Therefore, essentially consisting of a defined element excludes sources derived from separation and purification methods and pharmaceutically acceptable contaminants, such as: phosphate-buffered saline, which consists of "should be excluded from the composition of the invention Trace elements other than the essential method steps. Specific examples with definitions are within the scope of the present invention. Therefore: pH, temperature, time, concentration, and molecular weight are approximate 値, and the error can be in (+) or ( -) Interval 0.1. It is not a clear statement, but it is used before the index in this article. Although it is not a clear statement, the test described in this article is a lot of reagents known to be equivalent in the art. "Isolation" herein refers to separation Self-composed, cells and the compounds in them are usually related to natural objects. "Patient" or "host" refers to vertebrates, preferably dairy animals, and more preferably human diseases. Mammals include murine animals, monkeys, humans, Domestic animals, competitive animals, "cancer", "tumor" and "tumor" in this article can be used in singular or plural form, which means that they have a malignant transformation Diseased cells. Primary cancer cells (that is, cells taken from close sites) can be accepted by techniques (especially the listed elements and any composition other than method components). Trace amounts, preservatives, etc. "There are several other temporary definition terms when they are formed, such as the scope, which are understood, although" about ". It is also understood that the agents are only examples, and other parts, whether animal or mammal ( (But not limited to): and pets. It can be used for the purpose of malignant transformation of the host. It is distinguished from non-cancerous cells (-(7) (7) 200407119 test) and non-cancer cells. The definition of a cancer cell herein includes not only primary cancer cells, but also any cell derived from a cell ancestor. This includes metastatic cancer cells, as well as cultured cells in vitro and cell lines derived from cancer cells. When referring to this type of cancer usually refers to solid tumors, "clinically detectable" tumors are available; for example: computer tomography, magnetic resonance scanning (MRI), X-ray, ultrasound or palpation detection Tumor mass. Biochemical and immune findings alone do not fit this definition. "Inhibition" herein refers to stopping, delaying or slowing the growth, proliferation, or cell division or angiogenesis of endothelial cells in a tissue. Methods for monitoring inhibition include (but are not limited to): endothelial cell proliferation assay, blood content measurement, blood vessel bed volume, and quantitative determination of vascular structure density. When the cultured cells are mixed cells, the formation of new blood organs can be used to quantitatively measure the cells, showing the unique identity of endothelial cells, such as angiogenic factors, proteolytic enzymes, and endothelial cell-specific cell adhesion molecules to be monitored. "Composition" refers to a combination of an active agent with another inert and active compound or composition (such as a detectable agent or label), such as an adjuvant. "Pharmaceutical composition" is meant to include the active agent with an inert or A combination of active carriers to make a diagnostic or therapeutic composition for use in vitro, in vivo, or in vitro. "Pharmaceutically acceptable carrier" herein includes any standard pharmaceutical carrier, such as phosphate-buffered saline solution, water, and emulsions, such as oil / water or water / oil emulsions, and various types of wet化 剂。 Chemical agent. The composition may also include stabilizers and preservatives. For example, carriers, stabilizers, and Zo-11-(8) 200407119 agents, see Martin, R E MIN G TO N F S P H A R M. S D,] 5 1 (MackPubl. Co., Easton (1 975)). By "effective amount" is meant an amount sufficient to achieve a beneficial effect or desired result. For example, a therapeutic dose can achieve the desired therapeutic effect. This agent is the same or different from a dose effective to prevent the onset of disease or disease symptoms. An effective amount may be administered with one or more treatments, applications, or agents. As used herein, and unless otherwise described, "caproxen sulfur Alkane, is a "glyphosate" product that is degraded by sulfanil glucone. These ingredients are found in cruciferous vegetables (such as cabbage, broccoli, hard blue shoots, Brussels sprouts, broccoli, broccoli sprouts, Chinese cabbage, kale, Arugula, kohlrabi, mustard, radish, red sweet watercress). This term includes, unless otherwise described, the derivation of the vegetable sulphur compound and its pharmaceutically acceptable salts and prodrugs. And broccoli flower buds, there is a very rich glucosinolate glucosane derivative. The glucosinolate is also known as glucosylglucosane. The molecular formula of glucosinolate is C ^ H ^ NOSs and its molecular weight is 1 7 7.2 9. He is also known as "sulfur meals such as 4-methyl butane sulfite and (-)-1-isothiocyanines. One 4 (R)-" methyl thionine, but the courtyard may be It has anticancer properties because it can induce Period of detoxification, such as glutathione, S-transfer metabolite and benzophenone reduction metabolite. These anti-cancer properties are anti-carcinogens and toxic electrophilic substances. The applicant of the present invention has found that fortune sulforaphane can inhibit the endothelium Cells also have anti-angiogenic properties. According to such findings, the present invention can be dosed by the dose of hED.) The amount of "'cauliflower is usually broccoli, harvested and bio-specific glucose, and I ,,, : Can resist growth growth-12-(9) 200407119 Sending growth inhibition amount of vegetable sulfane, or a pharmacological substance, salt or prodrug to the cell can provide inhibitory endothelial button. The present invention also provides A method for inhibiting angiogenesis in tissues with pharmaceutically acceptable derivatives, salts or prodrugs that have good anti-angiogenic effects. This method can be performed in vitro or in vivo in vivo or in vivo when endothelial cells or vascularized Tissues can be cultured under conditions well known to those skilled in the art (for example, below). Tissues can be taken from cell lines established by themselves or cultured from samples taken from patient groups. The sulforaphane or its pharmacological properties can then be cultured. 'Salts or prodrugs added directly to culture medium or delivery of pharmaceutical. One dish turnip g sulfur derivatives of vegetable dish sulfane turnip turnip nitrous "unless specifically stated, embodiments comprising various separation. The name includes the racemic acid salt, and its purified formulae (D-, L-) and (D-, L), derivatives, such as, for example, pharmacologically acceptable salts of protease and prodrugs. The external purification method is as follows. One of its special features is the purified (D-) form. In another feature, it is only (Lfc) form. The (L-) form is a method in which each is commercially available and each is separated. For example, the use of HPLC for palmity, such as:, palmar-CBH and palmar-HAS, Tech Intr η ati ο na], AB · Perkins-E1 mer also has a separation kit, its name Sdfadex for palm separation group: acceptable method for derivation of 3 cell growth; sulfane, or its tissue, can be provided. Learn about the expertise of this skill in the living body. Cells and / or .Salts of acceptable derivatized composition of tissue sections. The specific implementation of this article is the application of the isomerized form of sulfanyl acetate, disulfide & sulphur-salted vegetables, and sulfanil is only racemic (D-) or palmitate-AGP Available from Chr OM for sale of palmar sets. (10) (10) 200407119 Implementations of this salt include: acetate, adipic acid, alginate, aspartate, benzene Formate, bisulfate, butyrate, citrate, camphor, camphor sulfonate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonic acid, trans Butenoate, glucoheptanoate, glyceryl phosphate, hemisulfate, heptanoate, caproate, hydrogen chloride, hydrogen bromide, hydrogen iodide, 2-fluorenylethanesulfonate, lactate, Maleate, mesylate, 2-naphthalenesulfonate, nicotinate, oxalate, palmitate, pectate, persulfate, phenylpropionate, picric acid H Salt, tert-valerate, propionate, succinate, tartrate, hydrosulfate 'tosylate and undecanoate. Examples of other salts include the compounds of the present invention Anions of substances such as: Na +, NHT, and NW 4 + (where W is a C 4 alkyl group). When used for treatment, the salts of the compounds of the present invention are salts accepted in medical class. However, non-medicine Scientifically acceptable salts of acids and bases can also be used, for example, to prepare or purify them into pharmaceutically acceptable compounds. Not every treatment is effective for each patient, so in vitro. The efficacy of the patient would be advantageous. Such methods are provided in the present invention to determine whether the treatment of calyxane can treat the specific disease associated with the pathological increase of endothelial cells in each patient. For example, it would be isolated from the patient The tissue living body is contacted with an effective amount of a medicinal composition and the therapeutic agent defined under conditions for effective cell growth and proliferation. The conventional method (eg, the CPAE assay described here) is used to determine the inhibition of pathological cell growth. It is shown that the composition and / or therapeutic agent of the present invention can effectively treat patients with disease. Angiogenesis or the formation of new blood vessels is the basic process of new blood vessels. -14-(11) (11) 200407119 process. They involve Physiological phenomena such as reproductive development and wound healing. Under normal circumstances, angiogenesis is highly regulated. However, many diseases are driven by persistent unregulated angiogenesis. In rheumatoid arthritis, New microvasculature invades joints and destroys cartilage. In diabetic retinopathy, new microvasculature invades the vitreous' in the retina, causing bleeding and blindness. Tumor growth and tumor metastasis are also interdependent with angiogenesis. Most primary solid tumors After a long period of no blood vessels (obvious latency), the maximum growth at this time is about 2 mm diameter. At this size, tumor cells can obtain the necessary oxygen and nutrients through simple passive diffusion. Such small tumor masses pass through Recruiting mature host blood vessels in the end can eventually open up angiogenesis and begin to grow new microvessels, eventually infiltrating into the tumor mass, thus causing the ruthless expansion of the tumor mass and bloody migratory walking. The onset of angiogenesis was initially triggered by a hypothetical foreign product and then subtly manipulated by the recipient's tumor cell growth factor, known as the "tumor angiogenesis factor" (TAF). The present invention also provides a method for treating patients with pathological neovascularization disease φ disease, which is administering to the patient an effective therapeutic amount of naphthyl sulfide · alkane (external nitrate), (D-) or (L ·) , Or a pharmaceutical composition containing one or more of these substances. "Treatment" herein refers to alleviating symptoms related to pathological neovascularization and / or endothelial cell growth and reducing neoangiogenesis or endothelial cell growth. The symptoms include (but are not limited to): arthritis symptoms, neovascular skin symptoms, diabetic retinopathy, Kaposi's sarcoma, aging macular degeneration, capillary dilatation, glaucoma, keloids, corneal transplant rejection Trauma granulomatosis, vascular fibrosis-15- (12) (12) 200407119 dimensional tumors, 0Ser-Webber syndrome, myocardial angiogenesis, and scleroderma. Exemplary arthritis symptoms are selected from: rheumatoid arthritis and osteoarthritis. In the treatment of cancer and solid tumors, "treatment includes the inhibition of blood vessel growth, causing tumors and / or cancer cells to lack the nutrients needed for their growth. Therefore tumors and growths will decrease in size and may disappear. Administration of medicines to treat arthritis symptoms, Will cause reduced cartilage (especially joint) blood vessel formation, resulting in increased mobility and flexibility in such areas. In the treatment of psoriasis, administration will reduce the symptoms on the skin, such as: pimple, flakes and visible blood vessels under the skin surface In diabetic retinopathy, the application of active lysate will reduce the formation of foreign blood vessels in the retina, resulting in unhindered vision. When treating Kaposi's sarcoma, the active lysate It can inhibit the growth and / or the further formation of blood vessels, thereby inhibiting the formation of damage and / or tumors. Active lysates are administered to patients, such as rats, rats or human patients, and medicines can be added to the medicine Academically acceptable carrier and for systemic, oral, transdermal or topical administration by the patient. Therapeutic amount can be used in the treatment method The toxicity of the lysate changes and changes. The active lysate can be delivered orally, intravenously, intraperitoneally, or percutaneously. When delivered to animals, this method is suitable for further confirmation of active lysate. Efficacy. In an example of an animal model, a group of nude mice (Balb / cNCRnu / nu females, Sim n η, G Π roy 5 CA) are each subcutaneously inoculated from about 105 to about 109 as defined herein After the transplantation is completed, the compound is administered, for example, injected subcutaneously near the transplantation site. The venier tester is used twice a week to determine whether the transplanted area is reduced. -16-(13) (13) 200407119 OK Proper use of MRL / lpr mice (MRL / MpJ-Fas5 Jackson Labs, Maine) to test or monitor the efficacy of arthritis symptoms. Benefits of positive treatment include reduced swelling of joints and hind feet of animals and reduced cartilage degradation (can be monitored by X-rays) A large group of Louis rats (8 weeks of age, 130-150 g, JACKSON LABS, MAINE, USA) received an immunoreactive injection of bovine cross-linked protein class II (B II) to produce rheumatoid disease. Soluble Emulsion of BII and ICFA (incomplete Freund's adjuvant) mixed with 0.1M acetic acid to 400 μg / ml (ug / mi) phase volume was injected at a concentration of 20 μg / 1 000 μl At the base of the rat's tail, when rheumatoid conditions occur, observe a series of physical symptoms and record the four-level classification within 28 days. Other animal models can also be used appropriately. The entire course of treatment can be Single dose, multiple doses, and continuous intermittent administration in vivo. The most effective method and method of administration are well known to those skilled in the art and will be based on the therapeutic composition, purpose of treatment, and target of treatment. Cell, and the patient being treated. Single and multiple administrations are possible, the dosage level and mode are selected by the treating physician. Appropriate dosage formulations and methods for administering reagents can be found below. The composition and the pharmaceutical preparation of the present invention can be used for preparing medicaments and health food supplements, and are administered according to conventional procedures (for example, a medical composition containing active ingredients) for treating humans and other animals. The pharmaceutical composition can be administered orally, intranasally ', parenterally or by inhalation, and can be in the form of tablets, lozenges, granules, capsules, medicine nine, ampoules, suppositories or aerosols. Water-soluble or water-insoluble active ingredients -17-(14) (14) 200407119 Diluents, syrups, granules or powders can also be in the form of suspensions, solutions and emulsions. In addition to the compound of the present invention, the pharmaceutical composition may contain other medically effective ingredients. More specifically, the active ingredients of the active lysate herein can be administered through any appropriate route during treatment, including oral, rectal, nasal, and coated (which includes transdermal , Aerosol, buccal, and sublingual), vaginal, parenteral (which includes subcutaneous, intramuscular, intravenous, and intradermal), and pulmonary routes of administration. The better path for φ will vary depending on the symptoms and age of the recipient, and the disease being treated. When treated in vivo, the patient's pathological endothelial cell growth or neovascularization is transplanted. It is the delivery of a therapeutically effective amount of calyxane, (racemate, D-, L-), or a pharmaceutically acceptable derivative, salt or prodrug thereof, to a patient to achieve the desired result. . The present invention also involves administering an effective therapeutic amount or a growth-suppressing amount of vegetable sulfane, (epilactate, D-, L,) or a pharmaceutically acceptable derivative, salt, or prolude Expulsion. Provide patients with a method for treating pathological neovascularization disorders. δH symptoms include (but are not limited to): arthritis symptoms, neovascular skin symptoms, diabetic retinopathy, Kaposi's sarcoma, aging macular degeneration, capillary dilatation, glaucoma, Keloids, corneal transplant rejection, traumatic granulomatosis, angiofibromas, Siei · -Webber syndrome, myocardial angiogenesis, and scleroderma. Exemplary arthritis symptoms are selected from: rheumatoid arthritis, psoriatic arthritis, and osteoarthritis. -18- (15) (15) 200407119 For patients, such as rats, rats or human patients, a pharmaceutically acceptable carrier can be added to the drug and used for systemic, oral, transdermal or local administration of the patient Apply to the patient. The amount of treatment can be determined experimentally and will vary depending on the pathological condition of the treatment, the patient treated, and the symptoms of the treatment. Not every individual treatment will be effective, so measuring the efficacy of individual patients in vitro would be an advantageous pre-step. Provided in the present invention are methods for determining whether such a composition or treatment can treat a patient's specific disease associated with the pathological proliferation or angiogenesis of endothelial cells. For example, 'isolating a tissue biopsy sample from a patient is contacted with an effective amount of a pharmaceutical composition as defined herein under conditions effective for cell growth and proliferative conditions. Using conventional methods, such as the CPAE measurement described herein, to measure the inhibition of pathological cell growth and growth, the results show that the composition and the therapeutic agent of the present invention can effectively treat patients. Although the music ingredient can be administered alone, it should preferably be administered as a blood formulation containing at least one effective ingredient (as defined above) with one or more pharmaceutically acceptable carriers and optionally other therapeutic agents. Each carrier must be "acceptable" and compatible with the other ingredients of the formulated species and not harmful to the patient. Preparations include oral, rectal, nasal, and coated (which include: percutaneous, cheek And sublingual), vaginal, parenteral (including subcutaneous, intramuscular, intravenous, and intradermal) and pulmonary preparations. Formulation methods can conveniently be in unit dosage form and can be prepared by any of the methods well-known on Witchcraft. The method includes the step of combining an active ingredient with a carrier to form one or more accessory ingredients. Generally, -19- (16) (16) 200407119 is used to prepare the preparation uniformly and tightly combining the active ingredient with a liquid carrier or a finely divided solid carrier or both, and then shape the product as necessary. The fatal preparations suitable for oral administration may be unlinked units (eg capsules, starch capsules or tablets) each containing a predetermined amount of active ingredient; it may be a 'powder or granule; a water-soluble or water-insoluble liquid solution Or suspension; or oil-in-water liquid emulsion or water-in-oil liquid emulsion. The active ingredient may also be a Chinese herbal medicine, a sugar medicine or a paste sauce. Tablets can be made by compression or molding, and can be considered to include one or more accessory ingredients. In the preparation of compressed tablets' the active ingredients in a loose form (such as powder or granules) can be compressed with a suitable machine, and binding agents (such as polyvinylpyrrolidone, gelatin, hydrocarbylpropylmethylcellulose), lubricants can be mixed if necessary. , Inert diluents, preservatives, disintegrating agents (such as: sodium starch glycolate, cross-linked polyvinylpyrrolidone, cross-linked methyl cellulose sodium) surface active or dispersing agents. Molded tablets can be made by moistening a powdery compound mixture with an inert liquid diluent using a suitable machine and molding. The tablets can be coated or dispersed as required, and the active ingredients can be formulated to provide sustained or controlled release formation, for example, various proportions of propylmethylcellulose can be used to provide the desired release form. The tablets may provide intestinal release as needed rather than in the stomach. Formulations suitable for topical administration in the oral cavity include: lozenges, which contain active ingredients commonly flavored with sucrose and gum arabic and targacons; the formulations may include active ingredients and inerts, such as gelatin And glycerin, or sucrose and gum arabic; and the preparation may be a mouthwash containing an active ingredient and a suitable liquid. The pharmaceutical composition for coating and administration according to the present invention can be made into a medicine. 20- (17) (17) 200407119 ointment, cream, suspension, coating, powder solution, ointment, 'spray, air gel or oily Thing. In addition, the preparation may contain a patch or dressing, such as a bandage or an adhesive plaster impregnated with the active ingredient and, if necessary, one or more excipients or diluents. When treating eyes or other external tissues, such as mouth and skin diseases, it is preferable to apply a coated ointment or cream preparation containing an active ingredient. When it is formulated into an ointment, the medicine can be prepared into an ointment using paraffin or a substance that can be mixed with water. In addition, the pharmaceutical ingredient can be formulated into an emulsifiable concentrate with an oil-in-water emulsifiable substance. If necessary, the aqueous phase of the emulsifiable concentrate includes, for example, at least about 30% w / w of a polyhydric alcohol, that is, an alcohol having two or more groups, such as propylene glycol, butane-1,3-diol, mannose Sugar alcohol, sorbitol, glycerin, and polyethylene glycol and mixtures thereof. The coating formulation may optionally include a compound that promotes absorption or penetration of the active ingredient of the drug through the skin or other infected areas. Examples of the real skin penetration enhancer include: dimethyl (yl) subseptum and related analogues. The oily phase of the emulsion of the present invention may be constituted in any conventional manner using conventional ingredients. This oil phase may contain only emulsifiers (i.e., conventional emulsifiers) and may optionally contain at least one emulsifier and a fat or oil or a mixture of fat and oil. Preferred hydrophilic emulsifiers may include lipophilic emulsifiers as stabilizers. The preferred ones may also include oils and fats. Emulsifiers with or without stabilizers can be made into so-called emulsifying waxes, and waxes and oils and / or fats can be made into so-called emulsifying ointments, which can form an oily dispersed phase of an emulsifiable concentrate. Suitable emulsifiers and emulsifier stabilizers for the present invention include:-~ (18) (18) 200407119 T, veen60, Span80, cetyl alcohol, myristyl alcohol, glyceryl monostearate, and lauryl Sodium sulfate. Since the active compound has very low solubility in most oils used in pharmaceutical emulsion preparations, it can be prepared by selecting an appropriate oil or fat according to the desired cosmetic properties. Therefore, preferred emulsifiable concentrates should have appropriate firmness to avoid leakage of non-greasy, non-staining, and washable products from the esophagus or other containers. Linear or branched, mono- or dibasic alkyl esters can be used, such as: di-isoadipate, isocetyl stearate, coconut fatty acid propylene glycol diester, isopropyl myristate , Oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate, or chain ester reference compounds, the conventional Crodamol CAP, the latter three being the preferred esters. These esters can be used alone or in combination depending on the nature. In addition, high melting point lipids can be used, such as white soft paraffin or liquid paraffin or other mineral oils. Formulations suitable for eye coating and administration, including active ingredients in eye drops that can be dissolved or suspended in a suitable carrier, especially Water-soluble solvents. Formulations for rectal administration may contain appropriate bases, which include, for example, a cocoa butter or salicylate suppository. Formulations suitable for vaginal administration may be vaginal suppositories, emboli, creams, gels, ointments, foams or spray formulations, which may contain suitable carriers known in the art in addition to the prodrug component. Formulations suitable for nasal administration, in which the carrier can be a solid including a coarse powder having a particle size of, for example, between about 20 to about 500 microns, which can be administered by inhalation, that is, from a powder containing powder near the nose. The container is quickly inhaled through the nasal cavity at -22-(19) (19) 200407119. Carriers suitable for, for example, nasal spray, nasal drip, or aerosol administration of a sprayer include a five-component water-soluble or liquid solution of the prodrug. Formulations suitable for parenteral administration include isotonic and water-insoluble isotonic solutions; sterilized injection solutions that can contain antioxidant oxidants (j, buffer solutions, bacteriostatic agents, and Blood isotonic solutes; and water-soluble and non-water-soluble sterilized suspensions, which may include suspending agents and thickeners, and microlipids or other designs to calibrate compounds to blood compositions or one or more An organ system is an example system. Preparations can exist in unit-dose or multi-dose sealed containers, such as ampoules and vials, and can be stored in a freeze-dried state, and only need to be added with a sterilized liquid carrier before use, For example, water can be used as an injection. Ready-to-use injection solutions and suspensions can be prepared from the previously described sterilized powders, granules, and tablets. Preferred unit dosage forms contain daily doses or units, and daily dosages (list (Above), or the pharmaceutical ingredients of its appropriate lysates. In addition to the above ingredients, the formulations of the present invention may include other ingredients that are known and adjusted in this art. Relevant various types of agents, such as suitable oral preparations, may further include: sweeteners, thickeners, and flavoring agents. They may also contain additional active agents, such as: anti-tumor, anti-cancer, Anti-angiogenic agent or immune enhancer. Prosthetic acid (racemic acid salt or optically pure combination), its prodrug, salt or its derivative and the same composition can also be used as a veterinary preparation It can be prepared, for example, by methods known in the art. -23- (20) (20) 200407119 The present invention further provides a method for co-selecting a therapeutic agent for inhibiting neovascularization or endothelial cell growth. The screening method comprises: (a ) Contacting the drug with an appropriate cell or tissue sample; (b) contacting the appropriate cell isolation sample or tissue sample with an effective treatment of Xia Zhicai Sulfur Institute or its medically acceptable derivatives, salts or prodrugs Contact, and (c) comparing the growth of the samples in steps (a) and (b), if the inhibitory effect of any agent on growth in step (a) is the same or similar to that in the sample in step (b), that is Inhibit new Therapeutic agents for tube formation or endothelial cell growth. Other treatments and agents described herein can be combined with samples as needed. The present invention also provides a kit for treating patients with pathological neovascularization disorders. The kit includes effective treatments Amounts of rapamidine (racemate, (D-) or (L0), or a pharmaceutically acceptable derivative, salt or prodrug thereof, and instructions. Conditions applicable to this set, It is selected from the group consisting of arthritis symptoms, neovascular skin symptoms, diabetic retinopathy, restenosis symptoms, Gabo's Kapose's sarcoma, aging macular degeneration, capillary dilatation, glaucoma, keloids, corneal transplant rejection , Granulomatous trauma, hemangiofibromas, os 1 er-Webber syndrome, myocardial angiogenesis, scleroderma rheumatoid arthritis, psoriatic arthritis, and osteoarthritis. The following examples are intended to illustrate, but not limit the invention. Example 1 24-(21) (21) 200407119 Example 1: Extraction and Purification of Phytosulfan is extracted and purified from the seeds of cruciferous plants. These methods are based on MATUSHESKI and colleagues () Eight §1 ^ 〇 :,? 〇〇 (^ 1 to 茁., 49; 1 8 67 -1 8 7 2). (D; L) -caprothane is synthesized from organic chemistry ((Kim , Sj Yi, (] 8 96). J. Org. Chem. 5], 26 1 3-2 6 1 5) D-caprosulfane is a racemic mixture of (D, L) -capro Sulfane is obtained using a palm column separation. All vegetable sulfanes can be purchased from LKTLABORATORIES

Example 2: Determination of Transplanted Meat Skin Cells of Brassinosulphane Measurement of Meat Skin Cells: The measurement of this skin cell was performed in a sentence CONALLY, etal. (1 9 8 6), ANAL · BI0CHEM, 152, 1 3 6 -4 0, and modified Method (LIANG AND WONG (1 9 99) ANGIOGENESIS: FROM THE MOLECULAR TO INTEGRATIVE PHARMACOLOGY, edited by MARADOUD-AKIS. Kluwes Academic / Plenm Pu NEW YORK), bovine CPAE cell line was purchased from American Type φ Culture Collection (ATCC), Cultured at a confluent state of MEM-10E to 95%. This cell can be prepared from a tissue culture flask using 0.25% trypsin solution, and then flattened on a 24 liter tissue culture plate with the same culture medium and having a density of 10, Q 0 0 cells per liter. After the plates were incubated at 37 ° C and 5% carbon dioxide gas for 8 hours, the test samples and controls can be added. Each 100 microliter sample was placed in two different wells for reproducibility. Guarantee. After incubating these samples for 60 hours, the medium solution can be absorbed, and then the number of cells can be determined by the acid phosphatase colorimetry method. Results and discussion:

Table I. L-caprothane inhibition of endothelial cell assays. L-Brazilian Sulfane Concentration (μg / ml) 0.78 μg / ml 1.56 μg / ml 3.13 μg / ml 6.25 μg / ml 12.5 μg / ml 25 μg / ml Inhibition of Endothelial Cell Growth (Percent) 0% 28.33% 71.38% 92.13% 94.5% 94.5% Table LLD- Asparagine sulfane inhibition endothelial cell assay. L-Brazilian Sulfane Concentration (μg / ml) 0.78 μg / ml 1.56 μg / ml 3.13 μg / ml 6.25 μg / ml 12.5 μg / ml 25 μg / ml Endothelial Cell Growth Inhibition (Percent) 0% 35.94% 74.13% 94.4 % 95.8% 95.6% -26- (23) 200407119 Table iu.d5l-Cabrasulfane inhibitory endothelial cell assay. Concentration of L-carrageenan (μg / ml) 0.78 μg / ml 1.56 μg / ml 3.13 μg / ml 6.25 μg / ml 12.5 μg / ml 25 μg / ml Inhibition of endothelial cell growth (percentage) 0% 31.85% 69.65% 92.72% 95.05% 95.08%

Each pair of calyxane can inhibit the growth of endothelial cells. The results of endothelial cell assays show that the addition of calyxane to the endothelial cell assay can effectively inhibit the growth of endothelial cells. At a concentration of 6.25 μg / ml, the growth of endothelial cells was inhibited by more than 92%. These results are precisely that the vegetable sulfane is a very effective inhibitor of endothelial cells. The measurements of the inhibition of angiogenesis by L-caprothane, D-caprothane, and D, L-caprothane at different Φ concentrations are shown in Figure 1. Vegetable sulfane is a very strong endothelial cell growth inhibitor even at very low concentrations. Example 3 Inhibition of a variety of cancer cell lines by caesium sulfane. These assays are based on the following steps: The cancer cell lines used include LncaP (prostate), SW4 80 (rectal) and HTB72 (melanoma), all purchased from AMERICAN TYPE CUTURE COLLECTION (ATTC), purchased, -27- (24) (24) 200407119 The cancer cells were placed in a 25 cm2 culture flask. Culture the medium of each cell to 90% confluent state. These cells are treated with trypsin, counted, and diluted to 1 500 cells per centiliter. One milliliter of cells is placed in a 24-well. Each well of a culture plate. Allow the cells to adhere overnight. Test samples at different concentrations were supplemented with 50 μl (u 1) water, p B S (phosphate buffered saline), or 1 X medium. Comparative sample wells will have the same volume of solution. The cells can grow for 60 hours at 37 ° C and 5% carbon dioxide gas. The culture medium of each well is aspirated, and the cells are washed with 1 ml of PBS and then started. Cell assay. 5000 microliters of substrate solution (1000 mg of molecular sodium acetate, ρΗ5.5, 0.2% Triton χ-]. 〇〇〇surfactant mg / ml p-nitrophenol phosphate) was added to the cell well Inside, the cell plate was incubated for 2 hours at 37.0, and then 1 ml of 0.1 mg (0.1 mg molecules) of sodium hydroxide was added. The optical density of these samples at 4 0 nm (microns) can be obtained from a micro plate器 测量。 Device determination. Results and discussion The inhibition of various cancer cell lines by L-caprothane has been listed in the following Tables IV to VI:-28- (25) 200407119 Table IV. Measurement of the growth inhibition of HTB72 cells by L-caprothane. Concentration of L. vegetable sulfane (μg / ml) 0.39 μg / ml 1.57 μg / ml 3.13 μg / ml 6.25 μg / ml 12.5 μg / ml 25 μg / ml HTB72 Cell growth inhibition percentage 17.21% 48.5% 89.6% 95.09% 95.83 % 96.3%

Table V. Measurement of Lncap cell growth inhibition by L-caprosulfane. Concentration of vegetable sulfane (μg / ml) 0.39 μg / ml] · 57 μg / ml 3.13 μg / ml 6.25 μg / ml 12.5 μg / ml 25 μg / ml Lncap Cell growth inhibition percentage 56.77% 74.22% 97.9% 97.9% 97.9 % 97.9% Table VI, Determination of L-Cabitrazine on SW4 8 0 Cell Growth Inhibition. O-Laptophan 0.39 1.57 3.13 6.25 12.5 25 Alkane concentration (picograms / mL mcg / μg / μg / μg / μg / G / ml) Milliliter Milliliter SW480 Cell growth inhibition percentage 15.13% 40.75% 83.27% 93.02% 94.3% 94.44%

-29- (26) (26) 200407119 We found that L-caprothane inhibits several cancer cell lines, including L11 cap (prostate cancer cell line) and SW 4 8 0 (rectal cancer cell line) and HTB72 ( Melanoma cancer cell line), L-caprothane at a very low concentration (6.25 μg / ml) has a strong inhibition on these various cancer cells, and this inhibition effect is especially effective against Ln cap (prostate cancer cell line). effective. Although the foregoing invention has been described in some detailed descriptions and embodiments for the sake of understanding, those skilled in the art can understand the present invention accordingly. The invention is altered and modified. For example, for those skilled in the art, the method of the present invention can be combined with one or more conventional anti-tumor, anti-angiogenic or immune-enhancing therapeutic agents and compositions, such as: sharks, shark cartilage, rokenan alcohol. Therefore, the above descriptions and examples should not be used to limit the true scope of the patent application scope of the present invention. [Schematic description] Figure 1 shows L-caproxane, D-vegetable service sulfur institute under endothelial cell assay test. And D'L-Inhibition of endothelial cell proliferation at different concentrations by TSSF. -30-

Claims (1)

(1) (1) 200407119 Patent application scope 1. A medicinal composition for inhibiting the growth of endothelial cells, comprising the delivery of a growth-suppressing amount of calyxane, or a pharmaceutically acceptable derivative or salt thereof Or prodrug to the cell. 2. The pharmaceutical composition according to item 1 of the patent application scope, wherein the vegetable sulfane is selected from the group consisting of: D-carved sulfane, L-carved sulfanilane, and D, L. . 3. For example, the pharmaceutical composition of the scope of application for patent 1, wherein the vegetable φ sulfane derivative is vegetable sulfan nitrite. 4. A pharmaceutical composition for inhibiting angiogenesis in a tissue, comprising delivering an antiangiogenic effect in an amount that is an antiangiogenic effect, or a pharmaceutically acceptable derivative, salt or prodrug thereof, to the tissue. 5. The pharmaceutical composition according to item 4 of the patent application scope, wherein the vegetable sulfane is selected from the group consisting of: D-carved sulfane, L-carved sulfane and D, L-carved sulfane racemate . 6. The pharmaceutical composition as claimed in item 4 of the patent application, wherein the vegetable sulfan # sulfane derivative is nitrite. 7. A pharmaceutical composition for inhibiting the angiogenesis of major organs, including the delivery of an anti-angiogenic effect (D-caprosulfane, caprosulfane, or cacosulfan racemate, or Its pharmaceutically acceptable derivative, salt or prodrug to the tissue. 8. The pharmaceutical composition according to any one of the claims 1, 4 or 7, wherein the delivery is in vitro, in vivo, or In vitro delivery of pharmaceutical composition. -31-(2) (2) 200407119 9 · A pharmaceutical composition for treating a condition related to angiogenesis in a pathological tissue in a host, including a dish for delivering an antiangiogenic effect Sulfanilane, or a pharmaceutically acceptable derivative, salt or prodrug thereof, to the tissue. 10. If the pharmaceutical composition of item 9 of the scope of the patent application, the vegetable sulfurium can be selected from the full portfolio including D- Vegetable sulphur compound, L-sulphur compound and D, L-sulphan acid racemic acid salt 11. The pharmaceutical composition according to item 9 of the patent application, wherein the disorder is selected from: tumors, joints Symptoms of inflammation, neovascular skin symptoms, diabetic retinopathy Kaposi's sarcoma, aging macular degeneration, restenosis, telangiectasia, glaucoma, keloids, corneal transplant rejection, traumatic granulomatosis, angiofibroma, Osier-Webber syndrome, myocardial angiogenesis, psoriasis arthritis, and crusts 12. The pharmaceutical composition according to item 9 of the scope of patent application, wherein the condition is selected from prostate cancer, rectal cancer and melanoma cancer. 13. The pharmaceutical composition according to item 9 of the scope of patent application, wherein the condition is Selected from the symptoms of arthritis of rheumatoid arthritis and osteoarthritis. 1 4. The pharmaceutical composition according to item 9 of the scope of application for patent, wherein the vegetable sulfane derivative is vegetable sulfane nitrite. 15 · — species A medicinal composition for treating a condition related to pathological neovascularization of a patient's main organ, comprising a therapeutically effective amount of D-caprothane, L-caprothane, and D, L-caprothionine Alkane, or a pharmaceutically acceptable derivative, salt or prodrug thereof.] 6 · If the pharmaceutical group of any one of the scope of application for patents Nos. 9 to 5 is -32- (3) 200407119, system Oral, intravenous, 'intraperitoneal, or subcutaneous administration. 1 7 · The pharmaceutical composition according to any one of claims 9 to 15 of the scope of patent application' further includes the administration of an effective amount of anti- Angiogenesis agent 18. The pharmaceutical composition according to item 16 of the application, wherein the host or patient is an animal. 19. The pharmaceutical composition according to item 17 of the scope of patent application, wherein the animal is selected from the group consisting of pets, livestock, and human patients. 2 0—A pharmaceutical composition that improves the general health and well-being of a patient, and contains an effective amount of sulforaphane administered to the patient. 2 1. The pharmaceutical composition according to item 20 of the patent application, which is delivered as a health food supplement. 22. The pharmaceutical composition of claim 20, wherein the host or patient is an animal. 23. The pharmaceutical composition according to item 21 of the patent application, wherein the animal is selected from the group consisting of pets, livestock and human patients. 24. The pharmaceutical composition according to item 20 of the patent application, wherein the vegetable sulfane is selected from the group consisting of D-caprothane, L-caproine and D, L-caproine. 2 5. The pharmaceutical composition according to item 20 of the patent application scope, wherein the vegetable sulfane derivative is vegetable sulfane nitrite. 2 6. —A pharmaceutical composition for screening a therapeutic agent for inhibiting neovascularization and endothelial cell growth, which comprises the following steps: a. Contacting the agent with an appropriate cell or tissue sample;-33 · (4) ( 4) 200 407 119 b. Contacting isolated samples of appropriate cell and tissue samples with the effective treatment of centrifugane and its pharmaceutically acceptable derivatives, salts or prodrugs; and c * comparison step (a) Similar to the growth of the sample in step (b), and in which the inhibitory effect of any agent in step (a) on the growth is the same and similar to that in the sample in step (b), it is a treatment that inhibits the formation of new blood vessels or endothelial cells Agent. 27. The pharmaceutical composition according to item 26 of the patent application, wherein the contact is in vitro or in vivo. 28. The pharmaceutical composition according to item 26 of the patent application scope, wherein the vegetable sulfane is selected from the group consisting of: D-vegetable sulfanil, L-vegetable sulfanil and D, L _ vegetable sulfan racemic Acid salt. 29. The pharmaceutical composition according to item 24 of the patent application scope, wherein the vegetable sulfane derivative is vegetable sulfane nitrite. 30. The pharmaceutical composition of claim 25, further comprising contacting the sample of steps a and b with an effective amount of an antiangiogenic agent. 3 1 * A package for treating patients with pathological neovascularization-related conditions, which comprises a therapeutically effective amount of naphthane, and its pharmaceutically acceptable derivatives, salts or prodrugs, and instructions for use . 3 2. The set according to item 31 of the scope of application for a patent, wherein the disorder is selected from the group consisting of cancerous tumors, arthritis symptoms, neovascular skin symptoms, diabetic retinopathy, Kap0Si, s sarcoma, and aging retina Macular degeneration, restenosis, telangiectasia, glaucoma, keloids, corneal transplantation -34-(5) (5) 200407119 Rejection, trauma granulomatosis, vascular 'fibroma, Osier-Webber syndrome, myocardial angiogenesis, and stiffness Dermatosis. 33. The kit according to item 31 of the scope of patent application, wherein the condition is selected from the group consisting of prostate cancer, rectal cancer and melanoma cancer. 34. The kit according to item 31 of the scope of patent application, wherein the condition is an arthritis symptom selected from rheumatoid arthritis and osteoarthritis. 3 5. The kit according to item 31 of the scope of patent application, wherein the vegetable sulfane is selected from the group consisting of: D-caprothane, L-caproline and D, L-caprothane racemic acid. salt. 36. The kit according to item 31 of the scope of patent application, wherein the derivatized sulfane derivative is derivatized sulfane nitrite. -35 >