Methods and Compositions Useful for Treating Cancer

Abstract

A cancer immunotherapy method for treating cancer in a patient comprising the steps of: removing lymphocytes from the peripheral blood of the patient; exposing the lymphocytes to one or more of i) a thymosin peptide, ii) a purified immune RNA or DNA. iii) one or more astragalus polysaccharides, iv) one or more letinous edodes polysaccharides, v) a cell wall component of Mycobacterium grannis, Mycobacterium phlei or Mycobaterium subtilis vi) a polysaccharide nucleic acid fraction of bacillus Calmette-Guerin, vii) a transfer factor viii) one or more non steroidal anti inflammatory agents and xi) an enkephalin chosen from methionine enkephalin or leucine enkephalin in vitro and infusing the exposed lymphocytes back into the patient is provided.

Claims

1. A composition comprising: i) a thymosin peptide, ii) a purified immune RNA or DNA. iii) one or more astragalus polysaccharides, iv) one or more letinous edodes polysaccharides, v) a cell wall component of Mycobacterium grannis, Mycobacterium phlei or Mycobaterium subtilis vi) a polysaccharide nucleic acid fraction of bacillus Calmette-Guerin vii) a transfer factor viii) one or more non steroidal anti inflammatory agents and ix) an enkephalin chosen from methionine enkephalin or leucine enkephalin.

2. The composition of claim 1, wherein the thymosin peptide is thymosin alpha 1 (TA1).

3. The composition of claim 1, wherein the enkephalin is methionine enkephalin.

4. The composition of claim 1, wherein the cell wall component is from Mycobacterium subtilis.

5. The composition of claim 1, wherein the non steroidal anti inflammatory agent is chosen from the group consisting of aceclofenac, acemetacin, amoxiprin, aspirin, azapropazone, benorilate, bromfenac, carprofen, celecoxib, choline magnesium salicylate, diclofenac, diflunisal, etodolac, etoracoxib, faislamine, fenbuten, fenoprofen, flurbiprofen, ibuprofen, indometacin, ketoprofen, ketorolac, lornoxicam, loxoprofen, lumiracoxib, meclofenamic acid, mefenamic acid, meloxicam, metamizole, methyl salicylate, magnesium salicylate, nabumetone, naproxen, nimesulide, oxyphenbutazone, parecoxib, phenylbutazone, piroxicam, salicyl salicylate, sulindac, sulfinprazone, suprofen, tenoxicam, tiaprofenic acid, and tolmetin.

6. The composition according to claim 5, wherein the non steroidal anti inflammatory agent is chosen from asprin and ibuprofen.

7. A cancer immunotherapy method for treating cancer in a patient, comprising the steps of: a) separating lymphocytes from the peripheral blood of the patient; b) exposing the lymphocytes to one or more of i) a thymosin peptide, ii) a purified immune RNA or DNA. iii) one or more astragalus polysaccharides, iv) one or more letinous edodes polysaccharides, v) a cell wall component of Mycobacterium grannis, Mycobacterium phlei or Mycobaterium subtilis vi) a polysaccharide nucleic acid fraction of bacillus Calmette-Guerin, vii) a transfer factor viii) one or more non steroidal anti inflammatory agents and xi) an enkephalin chosen from methionine enkephalin or leucine enkephalin in vitro and: c) infusing the exposed lymphocytes back into the patient.

8. The method of claim 7, wherein the removal step is performed by leukapheresis.

9. The method of claim 7, wherein the cancer is chosen from the group consisting of malignant tumors, benign tumors, solid tumors, sarcomas, carcinomas, hyperproliferative disorders, carcinoids, Ewing sarcomas, Kaposi sarcomas, brain tumors, tumors originating from the brain and/or the nervous system and/or the meninges, gliomas, glioblastomas, neuroblastomas, stomach cancer, kidney cancer, kidney cell carcinomas, prostate cancer, prostate carcinomas, connective tissue tumors, soft tissue sarcomas, pancreas tumors, liver tumors, head tumors, neck tumors, laryngeal cancer, esophageal cancer, thyroid cancer, osteosarcomas, retinoblastomas, thymoma, testicular cancer, lung cancer, lung adenocarcinoma, small cell lung carcinoma, bronchial carcinomas, breast cancer, intestinal cancer, colorectal tumors, colon carcinomas, rectum carcinomas, gynaecological tumors, ovary tumors/ovarian tumors, uterine cancer, cervical cancer, cervix carcinomas, cancer of body of uterus, corpus carcinomas, endometrial carcinomas, urinary bladder cancer, urogenital tract cancer, bladder cancer, skin cancer, epithelial tumors, squamous epithelial carcinoma, basaliomas, spinaliomas, melanomas, intraocular melanomas, leukemias, monocyte leukemia, chronic leukemias, chronic myelotic leukemia, chronic lymphatic leukemia, acute leukemias, acute myelotic leukemia, acute lymphatic leukemia and lymphomas.

10. The method of claim 7, wherein the cancer is liver cancer.

11. The method of claim 7, wherein the cancer is stomach cancer.

12. The method of claim 7, wherein the cancer is cervix carcinoma.

13. A method of treating cancer, comprising administering to a patient i) a thymosin peptide, ii) a purified immune RNA or DNA. iii) one or more astragalus polysaccharides, iv) one or more letinous edodes polysaccharides, v) a cell wall component of Mycobacterium grannis, Mycobacterium phlei or Mycobaterium subtilis vi) a polysaccharide nucleic acid fraction of bacillus Calmette-Guerin vii) a transfer factor viii) one or more non steroidal anti inflammatory agents and ix) an enkephalin chosen from methionine enkephalin or leucine enkephalin.

14. The method of claim 13, wherein the thymosin peptide is thymosin alpha 1 (TA1).

15. The method of claim 13, wherein the enkephalin is methionine enkephalin.

16. The method of claim 13, wherein the cell wall component is from Mycobacterium subtilis.

17. The composition of claim 13, wherein the non steroidal anti inflammatory agent is chosen from the group consisting of aceclofenac, acemetacin, amoxiprin, aspirin, azapropazone, benorilate, bromfenac, carprofen, celecoxib, choline magnesium salicylate, diclofenac, diflunisal, etodolac, etoracoxib, faislamine, fenbuten, fenoprofen, flurbiprofen, ibuprofen, indometacin, ketoprofen, ketorolac, lornoxicam, loxoprofen, lumiracoxib, meclofenamic acid, mefenamic acid, meloxicam, metamizole, methyl salicylate, magnesium salicylate, nabumetone, naproxen, nimesulide, oxyphenbutazone, parecoxib, phenylbutazone, piroxicam, salicyl salicylate, sulindac, sulfinprazone, suprofen, tenoxicam, tiaprofenic acid, and tolmetin.

18. The method of claim 13, wherein the cancer is chosen from the group consisting of malignant tumors, benign tumors, solid tumors, sarcomas, carcinomas, hyperproliferative disorders, carcinoids, Ewing sarcomas, Kaposi sarcomas, brain tumors, tumors originating from the brain and/or the nervous system and/or the meninges, gliomas, glioblastomas, neuroblastomas, stomach cancer, kidney cancer, kidney cell carcinomas, prostate cancer, prostate carcinomas, connective tissue tumors, soft tissue sarcomas, pancreas tumors, liver tumors, head tumors, neck tumors, laryngeal cancer, esophageal cancer, thyroid cancer, osteosarcomas, retinoblastomas, thymoma, testicular cancer, lung cancer, lung adenocarcinoma, small cell lung carcinoma, bronchial carcinomas, breast cancer, intestinal cancer, colorectal tumors, colon carcinomas, rectum carcinomas, gynaecological tumors, ovary tumors/ovarian tumors, uterine cancer, cervical cancer, cervix carcinomas, cancer of body of uterus, corpus carcinomas, endometrial carcinomas, urinary bladder cancer, urogenital tract cancer, bladder cancer, skin cancer, epithelial tumors, squamous epithelial carcinoma, basaliomas, spinaliomas, melanomas, intraocular melanomas, leukemias, monocyte leukemia, chronic leukemias, chronic myelotic leukemia, chronic lymphatic leukemia, acute leukemias, acute myelotic leukemia, acute lymphatic leukemia and lymphomas.

19. The method of claim 18, wherein the cancer is liver cancer.

20. The method of claim 18, wherein the cancer is stomach cancer.

21. The method of claim 18, wherein the cancer is cervix carcinoma.

22. The method of claim 13, wherein the purified immune RNA or DNA is in injection form.

23. The method of claim 13 wherein the cell wall component of Mycobacterium grannis, Mycobacterium phlei or Mycobaterium subtilis is in injection form.

24. The method of claim 23, wherein the cell wall component is from Mycobaterium subtilis.

25. The method of claim 13, wherein a polysaccharide nucleic acid fraction of bacillus Calmette-Guerin is in injection form.

26. The method of claim 13, wherein the enkephalin chosen from methionine enkephalin or leucine enkephalin is in injection form.

27. The method of claim 26, wherein the enkephalin is methionine enkephalin.

28. The method of claim 13, wherein the thymosin peptide, one or more astragalus polysaccharides, one or more letinous edodes polysaccharides and the transfer factor are each in tablet form.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0079] FIG. 1 shows a tumor animal model with S-180 cells. After administration of an IV injection with the composition of the invention, once a day for 7 successive days, the tumor size was measured. FIG. 1a shows a mouse tumor prior to treatment and FIG. 1b after treatment with a composition of the invention showing tumor shrinkage.

[0080] FIG. 2 shows the tumor size of S-180 carcinoma removed from mice before treatment with a preferred composition of the invention (FIG. 2a), after treatment with a preferred composition of the invention (FIG. 2b) and after treatment with a normal saline solution

[0081] FIG. 3a-d show dendritic cells before (FIGS. 3a and c) and after treatment (3b and d) with the composition visualized by scanning electronic microscope (SEM) to determine the maturation degree.

[0082] FIG. 4 shows an MRI of patient 1 before administration of a preferred composition of the invention (FIG. 4a) and after administration (FIG. 4b), showing tumor shrinkage

[0083] FIG. 5 shows an MRI of patient 2 before administration of a preferred composition of the invention (FIG. 5a) and after administration (FIG. 5b).

[0084] FIG. 6 shows an MRI of patient 3 before administration of a preferred composition of the invention (FIGS. 6a and 6c) and after administration (FIGS. 6b and 6d).

[0085] FIG. 7 shows an MRI of patient 4 before administration of a preferred composition of the invention (FIG. 7a) and after administration (FIG. 7b).

[0086] FIG. 8 shows stained blood cells of patient 5 before administration of a preferred composition of the invention (FIG. 8a) and after administration (FIG. 8b).

TERMS AND DEFINITIONS USED

[0087] The components of the formulation may be formulated for administration in any convenient way for use in human or veterinary medicine and the invention therefore includes within its scope pharmaceutical compositions comprising an ingredient of the invention adapted for use in human or veterinary medicine. Such compositions may be presented for use in a conventional manner with the aid of one or more suitable carriers. Acceptable carriers for therapeutic use are well-known in the pharmaceutical art, and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (A. R. Gennaro edit. 1985). The choice of pharmaceutical carrier can be selected with regard to the intended route of administration and standard pharmaceutical practice. The pharmaceutical compositions may comprise as, in addition to, the carrier any suitable binder(s), lubricant(s), suspending agent(s), coating agent(s), and/or solubilizing agent(s).

Routes of Administration and Unit Dosage Forms

[0088] The routes for administration include oral (e.g., as a tablet, capsule, or as an ingestible solution), topical, mucosal (e.g., as a nasal spray or aerosol for inhalation), nasal, parenteral (e.g., by an injectable form), gastrointestinal, intraspinal, intraperitoneal, intramuscular, intravenous, intrauterine, intraocular, intradermal, intracranial, intratracheal, intravaginal, intracerebroventricular, intracerebral, subcutaneous, ophthalmic (including intravitreal or intracameral), transdermal, rectal, buccal, epidural and sublingual. The compositions of the invention may be especially formulated for any of those administration routes. In preferred embodiments, the pharmaceutical compositions of the invention are formulated in a form that is suitable for oral delivery.

[0089] There may be different composition/formulation requirements depending on the different delivery systems. It is to be understood that not all of the compounds need to be administered by the same route. Likewise, if the composition comprises more than one active component, then those components may be administered by different routes. By way of example, the pharmaceutical composition of the invention may be formulated to be delivered using a mini-pump or by a mucosal route, for example, as a nasal spray or aerosol for inhalation or ingestible solution, or parenterally in which the composition is formulated by an injectable form, for delivery, by, for example, an intravenous, intramuscular or subcutaneous route. Alternatively, the formulation may be designed to be delivered by multiple routes.

[0090] Where the agent is to be delivered mucosally through the gastrointestinal mucosa, it should be able to remain stable during transit though the gastrointestinal tract; for example, it should be resistant to proteolytic degradation, stable at acid pH and resistant to the detergent effects of bile. For example, the compound of Formula I may be coated with an enteric coating layer. The enteric coating layer material may be dispersed or dissolved in either water or in a suitable organic solvent. As enteric coating layer polymers, one or more, separately or in combination, of the following can be used; e.g., solutions or dispersions of methacrylic acid copolymers, cellulose acetate phthalate, cellulose acetate butyrate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate, polyvinyl acetate phthalate, cellulose acetate trimellitate, carboxymethylethylcellulose, shellac or other suitable enteric coating layer polymer(s). For environmental reasons, an aqueous coating process may be preferred. In such aqueous processes methacrylic acid copolymers are most preferred.

[0091] When appropriate, the pharmaceutical compositions can be administered by inhalation, in the form of a suppository or pessary, topically in the form of a lotion, solution, cream, ointment or dusting powder, by use of a skin patch, orally in the form of tablets containing excipients such as starch or lactose, or in capsules or ovules either alone or in admixture with excipients, or in the form of elixirs, solutions or suspensions containing flavouring or colouring agents, or they can be injected parenterally, for example intravenously, intramuscularly or subcutaneously. For buccal or sublingual administration, the compositions may be administered in the form of tablets or lozenges, which can be formulated in a conventional manner.

[0092] When the composition of the invention is to be administered parenterally, such administration includes one or more of: intravenously, intraarterially, intraperitoneally, intrathecally, intraventricularly, intraurethrally, intrasternally, intracranially, intramuscularly or subcutaneously administering the agent; and/or by using infusion techniques.

[0093] Pharmaceutical compositions of the invention can be administered parenterally, e.g., by infusion or injection. Pharmaceutical compositions suitable for injection or infusion may be in the form of a sterile aqueous solution, a dispersion or a sterile powder that contains the active ingredient, adjusted, if necessary, for preparation of such a sterile solution or dispersion suitable for infusion or injection. This preparation may optionally be encapsulated into liposomes. In all cases, the final preparation must be sterile, liquid, and stable under production and storage conditions. To improve storage stability, such preparations may also contain a preservative to prevent the growth of microorganisms. Prevention of the action of micro-organisms can be achieved by the addition of various antibacterial and antifungal agents, e.g., paraben, chlorobutanol, or acsorbic acid. In many cases isotonic substances are recommended, e.g., sugars, buffers and sodium chloride to assure osmotic pressure similar to those of body fluids, particularly blood. Prolonged absorption of such injectable mixtures can be achieved by introduction of absorption-delaying agents, such as aluminium monostearate or gelatin.

[0094] Dispersions can be prepared in a liquid carrier or intermediate, such as glycerin, liquid polyethylene glycols, triacetin oils, and mixtures thereof. The liquid carrier or intermediate can be a solvent or liquid dispersive medium that contains, for example, water, ethanol, a polyol (e.g., glycerol, propylene glycol or the like), vegetable oils, non-toxic glycerine esters and suitable mixtures thereof. Suitable flowability may be maintained, by generation of liposomes, administration of a suitable particle size in the case of dispersions, or by the addition of surfactants.

[0095] For parenteral administration, the compound is best used in the form of a sterile aqueous solution, which may contain other substances, for example, enough salts or glucose to make the solution isotonic with blood. The aqueous solutions should be suitably buffered (preferably to a pH of from 3 to 9), if necessary. The preparation of suitable parenteral formulations under sterile conditions is readily accomplished by standard pharmaceutical techniques well-known to those skilled in the art.

[0096] Sterile injectable solutions can be prepared by mixing a compound of formula I with an appropriate solvent and one or more of the aforementioned carriers, followed by sterile filtering. In the case of sterile powders suitable for use in the preparation of sterile injectable solutions, preferable preparation methods include drying in vacuum and lyophilization, which provide powdery mixtures of the aldosterone receptor antagonists and desired excipients for subsequent preparation of sterile solutions.

[0097] The compounds according to the invention may be formulated for use in human or veterinary medicine by injection (e.g., by intravenous bolus injection or infusion or via intramuscular, subcutaneous or intrathecal routes) and may be presented in unit dose form, in ampoules, or other unit-dose containers, or in multi-dose containers, if necessary with an added preservative. The compositions for injection may be in the form of suspensions, solutions, or emulsions, in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, solubilizing and/or dispersing agents. Alternatively, the active ingredient may be in sterile powder form for reconstitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use.

[0098] The composition can be administered (e.g., orally or topically) in the form of tablets, capsules, ovules, elixirs, solutions or suspensions, which may contain flavoring or coloring agents, for immediate-, delayed-, modified-, sustained-, pulsed- or controlled-release applications.

[0099] The composition may also be presented for human or veterinary use in a form suitable for oral or buccal administration, for example in the form of solutions, gels, syrups, mouth washes or suspensions, or a dry powder for constitution with water or other suitable vehicle before use, optionally with flavoring and coloring agents. Solid compositions such as tablets, capsules, lozenges, pastilles, pills, boluses, powder, pastes, granules, bullets or premix preparations may also be used. Solid and liquid compositions for oral use may be prepared according to methods well-known in the art. Such compositions may also contain one or more pharmaceutically acceptable carriers and excipients which may be in solid or liquid form.

[0100] The tablets may contain excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate and glycine, disintegrants such as starch (preferably corn, potato or tapioca starch), sodium starch glycolate, croscarmellose sodium and certain complex silicates, and granulation binders such as polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin and acacia.

[0101] Additionally, lubricating agents such as magnesium stearate, stearic acid, glyceryl behenate and talc may be included.

[0102] The compositions may be administered orally, in the form of rapid or controlled release tablets, microparticles, mini tablets, capsules, sachets, and oral solutions or suspensions, or powders for the preparation thereof. In addition to the new solid-state forms of pantoprazole of the invention as the active substance, oral preparations may optionally include various standard pharmaceutical carriers and excipients, such as binders, fillers, buffers, lubricants, glidants, dyes, disintegrants, odourants, sweeteners, surfactants, mold release agents, antiadhesive agents and coatings. Some excipients may have multiple roles in the compositions, e.g., act as both binders and disintegrants.

[0103] Examples of pharmaceutically acceptable disintegrants for oral compositions include starch, pre-gelatinized starch, sodium starch glycolate, sodium carboxymethylcellulose, croscarmellose sodium, microcrystalline cellulose, alginates, resins, surfactants, effervescent compositions, aqueous aluminum silicates and cross-linked polyvinylpyrrolidone.

[0104] Examples of pharmaceutically acceptable binders for oral compositions include acacia; cellulose derivatives, such as methylcellulose, carboxymethylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose or hydroxyethylcellulose; gelatin, glucose, dextrose, xylitol, polymethacrylates, polyvinylpyrrolidone, sorbitol, starch, pre-gelatinized starch, tragacanth, xanthane resin, alginates, magnesium-aluminum silicate, polyethylene glycol or bentonite.

[0105] Examples of pharmaceutically acceptable fillers for oral compositions include lactose, anhydrolactose, lactose monohydrate, sucrose, dextrose, mannitol, sorbitol, starch, cellulose (particularly microcrystalline cellulose), dihydro- or anhydro-calcium phosphate, calcium carbonate and calcium sulphate.

[0106] Examples of pharmaceutically acceptable lubricants useful in the compositions of the invention include magnesium stearate, talc, polyethylene glycol, polymers of ethylene oxide, sodium lauryl sulphate, magnesium lauryl sulphate, sodium oleate, sodium stearyl fumarate, and colloidal silicon dioxide.

[0107] Examples of suitable pharmaceutically acceptable odourants for the oral compositions include synthetic aromas and natural aromatic oils such as extracts of oils, flowers, fruits (e.g., banana, apple, sour cherry, peach) and combinations thereof, and similar aromas. Their use depends on many factors, the most important being the organoleptic acceptability for the population that will be taking the pharmaceutical compositions.

[0108] Examples of suitable pharmaceutically acceptable dyes for the oral compositions include synthetic and natural dyes such as titanium dioxide, beta-carotene and extracts of grapefruit peel.

[0109] Examples of useful pharmaceutically acceptable coatings for the oral compositions, typically used to facilitate swallowing, modify the release properties, improve the appearance, and/or mask the taste of the compositions include hydroxypropylmethylcellulose, hydroxypropylcellulose and acrylate-methacrylate copolymers.

[0110] Examples of pharmaceutically acceptable sweeteners for the oral compositions include aspartame, saccharin, saccharin sodium, sodium cyclamate, xylitol, mannitol, sorbitol, lactose and sucrose.

[0111] Examples of pharmaceutically acceptable buffers include citric acid, sodium citrate, sodium bicarbonate, dibasic sodium phosphate, magnesium oxide, calcium carbonate and magnesium hydroxide.

[0112] Examples of pharmaceutically acceptable surfactants include sodium lauryl sulphate and polysorbates.

[0113] Solid compositions of a similar type may also be employed as fillers in gelatin capsules. Preferred excipients in this regard include lactose, starch, a cellulose, milk sugar or high molecular weight polyethylene glycols. For aqueous suspensions and/or elixirs, the agent may be combined with various sweetening or flavoring agents, coloring matter or dyes, with emulsifying and/or suspending agents and with diluents such as water, ethanol, propylene glycol and glycerin, and combinations thereof.

[0114] The composition may also, for example, be formulated as suppositories e.g., containing conventional suppository bases for use in human or veterinary medicine or as pessaries e.g., containing conventional pessary bases.

[0115] The composition may be formulated for topical administration, for use in human and veterinary medicine, in the form of ointments, creams, gels, hydrogels, lotions, solutions, shampoos, powders (including spray or dusting powders), pessaries, tampons, sprays, dips, aerosols, drops (e.g., eye ear or nose drops) or pour-ons.

[0116] For application topically to the skin, the agent of the invention can be formulated as a suitable ointment containing the active compound suspended or dissolved in, for example, a mixture with one or more of the following: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene polyoxypropylene compound, emulsifying wax, sorbitan monostearate, a polyethylene glycol, liquid paraffin, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, and water. Such compositions may also contain other pharmaceutically acceptable excipients, such as polymers, oils, liquid carriers, surfactants, buffers, preservatives, stabilizers, antioxidants, moisturizers, emollients, colourants, and odourants.

[0117] Examples of pharmaceutically acceptable polymers suitable for such topical compositions include acrylic polymers; cellulose derivatives, such as carboxymethylcellulose sodium, methylcellulose or hydroxypropylcellulose; natural polymers, such as alginates, tragacanth, pectin, xanthan and cytosan.

[0118] Examples of suitable pharmaceutically acceptable oils which are so useful include mineral oils, silicone oils, fatty acids, alcohols, and glycols.

[0119] Examples of suitable pharmaceutically acceptable liquid carriers include water, alcohols or glycols such as ethanol, isopropanol, propylene glycol, hexylene glycol, glycerol and polyethylene glycol, or mixtures thereof in which the pseudopolymorph is dissolved or dispersed, optionally with the addition of non-toxic anionic, cationic or non-ionic surfactants, and inorganic or organic buffers.

[0120] Examples of pharmaceutically acceptable preservatives include sodium benzoate, ascorbic acid, esters of p-hydroxybenzoic acid and various antibacterial and antifungal agents such as solvents, for example ethanol, propylene glycol, benzyl alcohol, chlorobutanol, quaternary ammonium salts, and parabens (such as methyl paraben, ethyl paraben and propyl paraben).

[0121] Examples of pharmaceutically acceptable stabilizers and antioxidants include ethylenediaminetetraacetic acid (EDTA), thiourea, tocopherol and butyl hydroxyanisole.

[0122] Examples of pharmaceutically acceptable moisturizers include glycerine, sorbitol, urea and polyethylene glycol.

[0123] Examples of pharmaceutically acceptable emollients include mineral oils, isopropyl myristate, and isopropyl palmitate.

[0124] The compounds may also be dermally or transdermally administered, for example, by use of a skin patch.

[0125] For ophthalmic use, the compounds can be formulated as micronized suspensions in isotonic, pH adjusted, sterile saline, or, preferably, as solutions in isotonic, pH adjusted, sterile saline, optionally in combination with a preservative such as a benzylalkonium chloride.

[0126] The pharmaceutical composition or unit dosage form of the invention may be administered according to a dosage and administration regimen defined by routine testing in the light of the guidelines given above in order to obtain optimal activity while minimizing toxicity or side effects for a particular patient. However, such fine tuning of the therapeutic regimen is routine in the light of the guidelines given herein.

[0127] The dosage of the active agents of the invention may vary according to a variety of factors such as underlying disease conditions, the individual's condition, weight, gender and age, and the mode of administration. An effective amount for treating a disorder can easily be determined by empirical methods known to those of ordinary skill in the art, for example by establishing a matrix of dosages and frequencies of administration and comparing a group of experimental units or subjects at each point in the matrix. The exact amount to be administered to a patient will vary depending on the state and severity of the disorder and the physical condition of the patient. A measurable amelioration of any symptom or parameter can be determined by a person skilled in the art or reported by the patient to the physician.

[0128] The pharmaceutical composition or unit dosage form may be administered in a single daily dose, or the total daily dosage may be administered in divided doses. In addition, co-administration or sequential administration of another compound for the treatment of the disorder may be desirable. To this purpose, the combined active principles are formulated into a simple dosage unit.

[0129] For combination treatment where the compounds are in separate dosage formulations, the compounds can be administered concurrently, or each can be administered at staggered intervals. For example, the compound of the invention may be administered in the morning and the antimuscarinic compound may be administered in the evening, or vice versa. Additional compounds may be administered at specific intervals too. The order of administration will depend upon a variety of factors including age, weight, gender and medical condition of the patient; the severity and aetiology of the disorders to be treated, the route of administration, the renal and hepatic function of the patient, the treatment history of the patient, and the responsiveness of the patient. Determination of the order of administration may be fine-tuned and such fine-tuning is routine in the light of the guidelines given herein.

DETAILED DESCRIPTION OF THE INVENTION

[0130] In accordance with the present invention, a cancer immunotherapy method for treating cancer in a patient comprising the steps of: [0131] a) separating lymphocytes from the peripheral blood of the patient; [0132] b) exposing the lymphocytes to one or more of i) a thymosin peptide, ii) a purified immune RNA or DNA. iii) one or more astragalus polysaccharides, iv) one or more letinous edodes polysaccharides, v) a cell wall component of Mycobacterium grannis, Mycobacterium phlei or Mycobaterium subtilis vi) a polysaccharide nucleic acid fraction of bacillus Calmette-Guerin, vii) a transfer factor viii) one or more non steroidal anti inflammatory agents and ix) an enkephalin chosen from methionine enkephalin or leucine enkephalin in vitro and: [0133] c) infusing the exposed lymphocytes back into the patient is provided.

[0134] The invention also provides a method of treating cancer comprising administering to a patient i) a thymosin peptide, ii) a purified immune RNA or DNA. iii) one or more astragalus polysaccharides, iv) one or more letinous edodes polysaccharides, v) a cell wall component of Mycobacterium grannis, Mycobacterium phlei or Mycobaterium subtilis vi) a polysaccharide nucleic acid fraction of bacillus Calmette-Guerin vii) a transfer factor viii) one or more non steroidal anti inflammatory agents and ix) an enkephalin chosen from methionine enkephalin or leucine enkephalin.

[0135] Alternatively, cancer patients can be treated through direct administration of agents mentioned above. The preferred composition, dosing regime and amounts are as follows:

(1) thymosin, IV or orally, 4 mg daily for two months;
(2) purified immune RNA, IV, 5 mg daily for two months;
(3) astragalus polysaccharide, IV or orally, 20 mg daily for two months;
(4) transfer factor, orally, 10 mg daily for two months;
(5) cell wall components, e.g. utilins, IV, 1 mg once a week for two months;
(6) polysaccharide of BCG, IV, three times, 2 mg in total, on days 1, 3, and 5;
(7) letinous edodes polysaccharide, IV or orally, 10 mg daily for two months;
(8) methionine encephalin, IV, 3 mg daily for three months; and
(9) ibuprofen, orally, 300 mg daily for 5 days.

[0136] The following examples provide a detailed illustration of the method of the present invention. These examples are not intended to limit or restrict the scope of the invention in any way.

[0137] The components of the composition invention are available from numerous commercial sources.

Examples

1. Impact on Lymphocytes Proliferation in Cancer Patients

[0138] 50 ml peripheral blood from a cancer patient was loaded on the upper layer of a Ficoll lymphocyte separation solution in a centrifuge tube and centrifuged at 1.500 rpm for 10 min. The white lymphocyte layer was removed with a pipette and the white lymphocyte layer was transferred cells were transferred into a flask. To the cells, the composition of the invention was added to dilute the cell number to 10.sup.5 cells/ml. The cells were cultured in an incubator at 37 C. for 24 h, whereby RPMI 1640 medium was used as a control. Increments of lymphocytes subpopulations where determined with flow cytometry using a BD Biosciences flow cytometer.

Average Percentage of Lymphocytes Subpopulations

[0139] After treating blood from 20 cancer patients with the composition of the invention, the following results were obtained using a BD Biosciences flow cytometer.

TABLE-US-00001 Control Treatment P CD3+ T cell 49.90 4.06 66.04 4.20 P < 0.01 CD8+ T cell 33.33 3.62 56.63 3.78 P < 0.01 CD4+ T cell 20.65 3.48 33.81 5.78 P < 0.01 NK cell 7.98 1.59 25.39 4.42 P < 0.01 Treg cell 4.62 0.64 2.85 0.48 P < 0.01 Total nucleated cell 1.10 0.13 4.01 0.75 P < 0.01

2. Testing Anticancer Activity of the Composition on Mice.

[0140] A tumor animal model was created with S-180 cells. An IV injection was administered with the preferred composition of the invention, once a day for 7 successive days and measure tumor size. Female C57BL/6 mice were obtained from Harlan SLAC Laboratory Animals Co. Ltd (Shanghai, China) (FIG. 1a). Mice were housed in the pathogen-free animal house at China Medical University and were used at 6-8 weeks of age. All experiments with animals were conducted in accordance with the Guide for the Care and Use of Laboratory Animals as adopted and promulgated by the China National Institutes of Health. Mice were inoculated with 10.sup.6 S180 cells subcutaneously to create a cancer model. After ending of treatment, the sizes of tumors were measured (FIGS. 1b and 2a-c).

3. Dendritic Cells Before and after Treatment with the Composition Separated dendritic cells were taken after treatment with the preferred composition of the invention followed by centrifugation and re-suspension in 0.5 ml 0.05 M pH 7.2 phosphorus buffer solution. The cells were then fixed in 2.5% glutaraldehyde, by adding 1% osmium tetroxide and allowing to stand overnight, dehydrated in ethanol and embedded in EPON resin. Sections were made on a Reiehert-Jung Ultracut E, stained with uranyl acetate and lead citrate. The cells were then viewed using a scanning electron microscope to determine degree of maturation (FIGS. 3a-d).
4. Treatment of Cancer Patients with the Composition Using Leukapharesis

[0141] Regime: Before treatment all cancer patients underwent CT scans (GECT/e USA). Cancer patients were then administered with the invention through iv or orally, every day for one month, from second month on taking the preferred composition of the invention every other day for another one month:

(1) thymosin, IV or orally, 4 mg daily for two months;
(2) purified immune RNA, IV, 5 mg daily for two months;
(3) astragalus polysaccharide, IV or orally, 20 mg daily for two months;
(4) transfer factor, orally, 10 mg daily for two months;
(5) cell wall components, e.g. utilins, IV, 1 mg once a week for two months;
(6) polysaccharide of BCG, IV, three times, 2 m (correct???YES), on days 1, 3, and 5;
(7) letinous edodes polysaccharide, IV or orally, 10 mg daily for two months;
(8) methionine encephalin, IV, 3 mg daily for three months; and
(9) ibuprofen, orally, 300 mg daily for 5 days.

[0142] After ending of treatment for two months, a CT scan was taken again and at the same time lymphocytes sub-populations were measured with flow cytometry.

[0143] Patient 1: a 55 year old female, with terminal lung cancer, treated with a composition of the invention via leukopharesis (white cells separated from blood and exposed to a preferred composition of the invention and reintroduced to the patient) four times in two months and by administration of a preferred composition using the preferred dosing:

(1) thymosin, IV or orally, 4 mg daily for two months;
(2) purified immune RNA, IV, 5 mg daily for two months;
(3) astragalus polysaccharide, IV or orally, 20 mg daily for two months;
(4) transfer factor, orally, 10 mg daily for two months;
(5) cell wall components, e.g. utilins, IV, 1 mg once a week for two months;
(6) polysaccharide of BCG, IV, three times, 2 mg in total on days 1, 3, and 5;
(7) letinous edodes polysaccharide IV or ora mg daily for two months;
(8) methionine encephalin, IV, 3 mg daily for three months; and
(9) ibuprofen, orally, 300 mg daily for 5 days.

[0144] Tumor shrinkage was observed as evidenced by the MRI shown in FIG. 4.

[0145] Patient a 56 year old male, with severe terminal liver cancer metastasized from rectal cancer treated with a composition of the invention leukopharesis (white cells separated from blood and exposed to a preferred composition of the invention and reintroduced to the patient) four times in two months and by administration of a preferred composition using the preferred dosing. Tumor shrinkage was observed as evidenced by the MRI in FIG. 5.

[0146] Patient 3: a 47 year old female with terminal lymphoma, treated with a composition of the invention via leukapharesis (white cells separated from blood and exposed to a preferred composition of the invention and reintroduced to the patient) four times in two months and by administration of a preferred composition using the preferred dosing. Tumor shrinkage observed as evidenced by the MRI shown in FIG. 6.

[0147] Patient 4: a 57 year old male with severe terminal liver cancer, treated with leukapharesis (white cells separated from blood and exposed to a preferred composition of the invention and reintroduced to the patient) using the composition of the invention four times in two months and by administration of a preferred composition using the preferred dosing. Tumor shrinkage was observed as evidenced by the MRI shown in FIG. 7.

[0148] Patient 5 a 25 year old male with severe terminal leukemia was treated with leukapharesis (white cells separated from blood and exposed to a preferred composition of the invention and reintroduced to the patient) with the composition four times in two months and by administration of a preferred composition using the preferred dosing. Tumor cells returned to normal as evidenced by microscopic examination shown in FIG. 8.

[0149] Patient 6 a woman, 53, with terminal cervical cancer and metastasized to whole body and with ascites. Post treatment with agents above for three months the patient removed and all ascites disappeared. Immune system returned to normal.

[0150] Patient 7 a man 66, with terminal stomach sinus cancer and digestive track blockage caused by cancer. Patient was treated with a preferred composition of the for four months with tumor shrinkage observed.