Pharmaceutical compositions and methods for treatment of intraepithelial dysplasia and neoplasia

Abstract

Pharmaceutical compositions comprising active doses of EGCG or a combination of EGCG and quinine or EGCG and a zinc salt for topical treatment of dysplastic or neoplastic cells in mucosal or cutaneous epithelial cells, including cervical intra-epithelial neoplasia, and methods for their use to induce regression of mucosal dysplasia are provided.

Description

[0001] PHARMACEUTICAL COMPOSITIONS AND METHODS FOR TREATMENT OF INTRAEPITHELIAL DYSPLASIA AND NEOPLASIA

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Application No. 63 / 639,422, filed April 26, 2024. The contents of the aforementioned application are hereby incorporated by reference in their entirety.

[0004] BACKGROUND

[0005] The present invention is in the technical field of cancer prevention and treatment. More particularly, the present invention is in the technical field of topical treatment of precancerous and early cancerous lesions of mucosal and cutaneous epithelium. Most cases of cervical cancer are initiated by infection of cells with Human Papillomavirus (HPV); the same is true of other mucosal epithelial cancers, including vulvar, vaginal, anal and oropharyngeal malignancies. Cervical squamous epithelial abnormalities are graded from low to high, the lowest identifiable fomi being Atypical Squamous Cells (ASC), which is divided into two groups: Undetermined Significance (ASCUS) and ‘“Cannot exclude HSIL’ (ASC-H), followed by Cervical Intraepithelial Neoplasia 1 (C1N 1) to higher grades, C1N2 and CIN3, using the CIN classification system. Using The 2014 Bethesda System (TBS), the terminology “Low-Grade Squamous Intraepithelial Lesions (LSIL)” to “High-Grade Squamous Intraepithelial Lesions (HSIL)” are employed. Beyond CIN3 or HSIL lesions, squamous cell abnormalities are either described as having “features suspicious for invasion” or Squamous Cell Carcinoma. Glandular abnonnalities can be described using either system. The umbrella term for such cervical epithelial cell abnormalities, whether squamous or glandular, is “cervical dysplasia”. Cervical dysplasia can be diagnosed several ways, including by naked eye visual examination using acetic acid (visual inspection with acetic acid (VIA)) or Lugol’s iodine (visual inspection with Lugol’s iodine (VILI)) or with the assistance of an intravaginal microscope known as a colposcopc. In certain cases, diagnostic biopsies arc taken and assessed via histology, at which point, the CIN classification system is used. Cervical dysplasia can also diagnosed using exfoliative cytology (Papanicolaou Test, “Pap smear”, or “Pap test”), in which a scraping of cervical epithelial cells are collected and analyzed using appropriate staining methods. Cervical dysplasia diagnosed by cytology is graded using The Bethesda System (TBS) where LSIL (encompassing HPV / mild dysplasia) is an appropriate surrogate for CIN 1 , and HSIL (encompassing moderate and severe dysplasia / Carcinoma in Situ) an appropriate surrogate for CIN2 / 3. Low-grade dysplastic lesions (e.g. ASCUS. CIN1 / LSIL) are often managed by an observational approach, requiring repeat cervical examination known colloquially as “watching -and-waiting”. Despite the fact that a large proportion of ASCUS and CIN1 / LSIL lesions do regress naturally typically within 2-5 years, the diagnosis itself imposes fear and psychosocial stress during the waiting period. Nonetheless, low-grade abnormalities carry the risk of progress toward malignancy, but existing surgical and ablative treatments can damage the cervix, compromising the ability to have a successful pregnancy, and are therefore not considered warranted for ASCUS and CIN1, and undesirable for CIN2. However, as almost all cervical cancer begins as low-grade abnormalities, a relatively benign method for eliminating dysplastic cells would have a large impact on the risk of developing cervical cancer in individuals and on its incidence in populations. This is especially important in low-resource settings where cervical dysplasia screening and treatment are not widespread or affordable.

[0006] Tire present invention is in the technical field of treatinent of dysplasia and neoplasia in mucosal and cutaneous epithelium. More particularly, the present invention is in the technical field of topical therapy of epithelial neoplasia. More particularly, the present invention is in the technical field of pharmaceutical compositions, especially intravaginal suppositories, that promote regression of epithelial neoplasia, including cervical dysplasia, genital, oral and anal mucosal neoplasia, and warts, including genital warts.

[0007] The earliest stages of epithelial dysplasia in the cervix, ASCUS and CIN1, are generally not treated, and are simply observed and graded during routine cervical examination using a combination of Papanicolau (Pap) tests, molecular diagnostic testing to determine the presence of HPV (particularly high- risk HPV strains (HR-HPV) which cause cervical dysplasia), and via visual examination using staining techniques during colposcopy (intravaginal microscopy) with or without the inclusion of diagnostic biopsies, given that a substantial fraction may regress unaided over time, and current treatment modalities cause damage to the cervix that may compromise the ability to become pregnant or carry a pregnancy to term. If CIN 1 progresses to CIN2 or CIN3, presenting a greater risk of transition of malignant cervical cancer, the most common treatments are surgical ablation, including LEEP (loop electrosurgical excision procedure) and thermal ablation, involving destruction of dysplastic regions and associated margins of nontransformed epithelial cells with a heated probe, and laser conization, also involving epithelial destruction with margins. Both surgical and ablative techniques result in consequent pain and risk of pregnancy-impairing damage. Both excisional and ablative surgical treatments require an average of two - six weeks post-procedure to heal, may cause ongoing discomfort, and generally cannot be repeated for recurrent dysplasia due to excessive cervical damage. Surgical procedures require local or general anesthetic, and run the risk of causing infections, bleeding, scarring to the cervix, and may compromise the ability to become pregnant and carry a pregnancy to term. Thermal ablation similarly damages normal tissues surrounding areas of dysplasia, causes cramping, and the combination of smoke generated by the procedure and smell of cauterized mucosal tissue is unpleasant and can cause distress to the patient. Surgical procedures also require extensive operator expertise, years of training, and associated certification to perform successfully, resulting in limits to access to appropriate treatment for cervical dysplasia. Cervical procedures frequently induce a vagal response which can lead to dizziness, fainting, nausea, ringing ears, and sweating.

[0008] There exists a need for a relatively benign topical treatment capable of inducing regression of ASCUS. CIN1, CIN2 or CIN3 to lower grades of dysplasia or normal status, thereby reducing risk of transition to cervical cancer, which would have a large impact on both personal health and upon healthcare costs for populations. A treatment that can be self-administered would be especially advantageous.

[0009] SUMMARY

[0010] The present invention comprises novel compositions for topical therapy of epithelial dysplasia and neoplasia, including but not limited to cervical neoplasia, vulvar, oropharyngeal and anal dysplasia, and genital warts.

[0011] The present invention provides compositions comprising EGCG (epigallocatechin gallate, a polyphenol extracted from green tea) alone as an active agent, or in combination with quinine as dual active agents, for treatment of epithelial dysplasia, including all stages of cervical dysplasia, vulvar, vaginal, anal and oropharyngeal dysplasia and neoplasia, genital warts, and cutaneous precancerous and cancerous lesions, including actinic keratosis, and some squamous cell cancers. The invention comprises novel pharmaceutical compositions, including combinations of active agents with unexpected synergistic activity, and which overcome stability problems, fonnulations for topical delivery of the active agents to sites of epithelial dysplasia.

[0012] In one embodiment for treatment of cervical epithelial dysplasia, the invention provides intravaginal suppositories that can be self-administered for a brief treatment period to promote regression of cervical dysplasia toward lower grades, including elimination of dysplasia and subsequent HPV- ncgativc molecular diagnostics.

[0013] Enumerated Embodiments

[0014] 1. A pharmaceutical composition comprising epigallocatechin gallate (EGCG), wherein the pharmaceutical composition is formulated as a suppository or atopical ointment, cream, or gel.

[0015] 2. A pharmaceutical composition comprising epigallocatechin gallate (EGCG), which lacks an EGCG degradant, or which comprises the EGCG degradant at a level less than 50%, 40%, 30%, 20%, 10%, 5%, 3%, 2%, or 1% of the EGCG, optionally wherein the pharmacal composition is fonnulated as a suppository or atopical ointment, cream, or gel. 3. A pharmaceutical composition comprising epigallocatechin gallate (EGCG), in a dosage form comprising 25-75 mg, 30-70 mg, 40-60 mg, or 45-55 mg, e.g., 50 mg, or 225-500 mg of tire EGCG.

[0016] 4. A pharmaceutical composition comprising EGCG, wherein the pharmaceutical composition lacks an antioxidant or lacks an excipient with antioxidant or reducing activity other than EGCG, or comprises an antioxidant or an excipient with antioxidant or reducing activity other than EGCG at a level of less than 5, 4, 3, 2. or 1 mg per dosage form (e.g., less than 2.5, 2, 1.5, 1, or 0.5 mg of antioxidant or excipient with antioxidant or reducing activity / g of suppository).

[0017] 5. The pharmaceutical composition of embodiment 4, wherein the antioxidant other than EGCG or a salt thereof is astaxanthin, and the astaxanthin is present at a level of less than 5, 4, 3, 2, or 1 mg per dosage form (e.g., less than 2.5, 2, 1.5, 1, or 0.5 mg / ml).

[0018] 6. A pharmaceutical composition comprising EGCG, wherein the pharmaceutical composition lacks a reducing agent other than EGCG, or comprises a reducing agent other than EGCG at a level of less than 5, 4. 3, 2, or 1 mg per dosage form (e.g.. less than 2.5. 2, 1.5, 1, or 0.5 mg / ml).

[0019] 7. The pharmaceutical composition of any of the preceding embodiments, which further comprises quinine or a salt thereof (e.g., quinine hydrochloride, quinine sulfate, or quinine sulfate dihydrate), quinine congeners and metabolites, quinidine, synthetic quinoline compounds, chloroquine, hydroxychloroquine, mefloquine, primaquine, quinacrine, and / or acridine orange.

[0020] 8. Tire pharmaceutical composition of any of the preceding embodiments, which further comprises zinc or a salt thereof, e.g., zinc acetate, zinc gluconate, or zinc citrate.

[0021] 9. The pharmaceutical composition of any of the preceding embodiments, which further comprises sucrose or a salt thereof.

[0022] 10. The pharmaceutical composition of any of the preceding embodiments, which further comprises citric acid (e.g., citric acid monohydrate) or a salt thereof.

[0023] 11. The pharmaceutical composition of any of the preceding embodiments, wherein:

[0024] (i) the EGCG is present at an equal amount by weight as the quinine or salt thereof;

[0025] (ii) the EGCG is present at a higher amount by weight than the zinc or salt thereof;

[0026] (iii) the quinine or salt thereof is present at a higher amount by weight than the zinc or salt thereof; and / or

[0027] (iv) the ratio of the EGCG to quinine or salt thereof to zinc or salt thereof is 10 + / - 20%: 10 + / - 20% : 1 + / - 20%. 12. The pharmaceutical composition of any of the preceding embodiments, wherein:

[0028] (i) the EGCG is present at 1-10%, e.g., 1.5-8.5%, e.g., 2-6%, e.g., 2.5-5%, e.g., about 2.5% or about 5% w / w;

[0029] (ii) the quinine or the salt thereof is present at 1-10%, e.g., 1.5-8.5%, e.g., 2-6%, e.g., 2.5-5%, e.g., about 2.5% or about 5%w / w;

[0030] (iii) the citric acid (e.g., citric acid monohydrate) is present at 0.1-1%. e.g., 0.2-0.8%, e.g., 0.3- 0.7%, e.g., 0.4-0.6%, e.g., about 0.5% w / w;

[0031] (iv) the sucrose is present at 1-4%, e.g., 1.5-3.5%, e.g., 2-3%, e.g., about 2.5% w / w; and / or

[0032] (v) the zinc (e.g., zinc acetate) is present at 0.01-1%, e.g., 0.1-0.9%, e.g., 0.1-0.5%, e.g., about 0.2%, e.g., 0.24% w / w.

[0033] 13. The pharmaceutical composition of any of the preceding embodiments, comprising one or more (e.g., 1, 2, 3, 4, or 5) excipients, wherein the excipients comprise PEG300, PEG300 stearate (e.g., PEG300 monostearate), PEG400, PEG400 stearate (e.g., PEG400 monostearate), PEG1500, PEG1500 stearate (e.g., PEG1500 monostearate), PEG3350, PEG3350 stearate (e.g., PEG3350 monostearate) and / or HPMC (e.g., HPMC KI OOM).

[0034] 14. The pharmaceutical composition of any of the preceding embodiments, comprising one or more (e.g., 1, 2, 3, 4, 5, or more) PEG stearates, e.g., one or more PEG monostearates comprising a PEG having a molecular weight in the range of 200-2000, e.g., one or more PEG monostearates comprising a PEG having a molecular weight in the range of 300-1500.

[0035] 15. The pharmaceutical composition of any or the preceding embodiments, comprising:

[0036] (a) 1-10%, e.g., 1.5-8.5%. e.g., 2-6%, e.g., 2.5-5%, e.g.. about 2.5% or about 5% w / w quinine or salt thereof, e.g., quinine sulfate:

[0037] (b) 1-10%, e.g., 1.5-8.5%, e.g., 2-6%, e.g., 2.5-5%, e.g., about 2.5% or about 5% w / w EGCG;

[0038] (c) 12.5-27.5%, e.g., 15-25%, e.g., 17.5-22.5%, e.g., about 20% w / w PEG 400 monostearate;

[0039] (d)

[0040] (i) 15-35%, e.g., 20-30%, e.g., 22.5-27.5%, e.g., about 25%, e.g., 25.16% w / w PEG 400; or

[0041] (ii) a composition comprising PEG stearate, e.g., PEG300 monostearate and PEG1500 monostearate, e.g., 0.01-99.9% PEG300 monostearate and 0.01-99.9% PEG1500 monostearate, e.g., Tcfosc® 1500;

[0042] (e) 30-50%, e.g., 35-45%, e.g., 37.5-42.5%, e.g., about 40% w / w PEG 3350; (f) 0.1-1%, e.g., 0.2-0.8%, e.g., 0.3-0.7%, e.g., 0.4-0.6%, e.g., about 0.5% w / w citric acid, e.g., citric acid monohydrate;

[0043] (g) 1-4%, e.g., 1.5-3.5%, e.g., 2-3%, e.g., about 2.5% w / w sucrose;

[0044] (h) 0.01-1%, e.g., 0.1-0.9%, e.g., 0.1-0.5%, e.g., about 0.2%, e.g., 0.24% w / w zinc acetate; and

[0045] (i) 2.5-12.5%, e.g., 5-10%, e.g., 6-9%, e.g., about 7%. e.g., 6.6% w / w HPMC. e.g., HPMC K100M.

[0046] 16. The pharmaceutical composition of any of the preceding embodiments, which is free of, is substantially free of, or comprises less than:

[0047] (i) 75%, 50%, 25%, 10%, 1%. 0.1%, or 0.01% by weight of Sanguinaria canadensis (bloodroot); and / or

[0048] (ii) 1%, 0.1%. or 0.01% by weight of Hydrastis, e.g., Hydrastis tincture, thuja oil. tea tree oil, bitter orange oil, vitamins (e.g., vitamin A), iron, ferric sulfate, ferrous sulfate, green tea extract, catechins other than EGCG (e.g., (-)-epicatechin (EC), (-)-epicatechin-3-gallate (ECG), (-)-epigallocatechin (EGC), gallocatechin, catechin gallate, gallocatechin gallate), and / or theasinensin A.

[0049] 17. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the pharmaceutical composition is formulated as a suppository.

[0050] 18. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the pharmaceutical composition, suppository , or topical ointment, cream, or gel is formulated for:

[0051] (i) vaginal formulation;

[0052] (ii) anal administration;

[0053] (iii) topical administration; or

[0054] (iv) oral administration.

[0055] 19. A method of treating or preventing a cancer, pre-cancer, or infection in a subject in need thereof, comprising administering the pharmaceutical composition of any of the previous embodiments to a subject.

[0056] 20. The method of embodiment 19, wherein the cancer, pre-cancer, or infection is, or is identified as, a mucosal epithelial cancer (e.g., vulvar, vaginal, anal, or oropharyngeal malignancy), a surface dysplasia (e.g., vulvar, anal, inguinal, oropharyngeal), an epithelial neoplasia (e.g., cervical dysplasia, genital mucosal neoplasia (e.g., vulvar epithelial dysplasia), oral mucosal neoplasia, oropharyngeal dysplasia, oropharyngeal neoplasia, or anal mucosal neoplasia), cutaneous precancerous and cancerous lesions (e.g., actinic keratosis), a squamous cell cancer, a glandular cell cancer, warts (e.g., genital warts, e.g., HPV- induced external genital warts), cervical cancer, or HPV infection.

[0057] 21. The method of embodiments 19 or 20. wherein the subject is infected, or is identified as infected, with HPV 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, or 59.

[0058] 22. The pharmaceutical composition of any of the preceding embodiments, which is an ointment, cream, or gel.

[0059] 23. The pharmacal composition of embodiment 22, which comprises a stabilizing excipient.

[0060] 24. The pharmaceutical composition of embodiment 22 or 23, which comprises an organic acid (e.g., citric acid), zinc or a pharmaceutically acceptable salt thereof (e.g., zinc acetate), and / or sucrose.

[0061] 25. The pharmaceutical composition of any of embodiments 22-24, which comprises a ratio of oil to water of 85: 15, 80:20, 75:25, 60:40, 55:45, 50:50, 45:55, or 40:60.

[0062] 26. The pharmaceutical composition of any of embodiments 22-25, which comprises a first PEG (e.g., PEG 4000) and a second PEG (e.g., PEG 400).

[0063] 27. The pharmacal composition of embodiment 26, wherein the first PEG and the second PEG have a molecular weight of 200-6000, e.g., 200-500, 500-1000, 1500-2000, 2500-3000, 3500-4000, 4500-5000, or 5500-6000.

[0064] 28. The pharmaceutical composition of embodiments 26 or 27, wherein the first PEG and the second PEG have different molecular weights.

[0065] 29. The pharmaceutical composition of any of embodiments 22-28, which comprises 60-80% (e.g., 80%, 75%, 70%, 65%, or 60%) PEG 400 and 20-40% (e.g., 20%, 25%, 30%, 35%, or 40%) PEG 4000.

[0066] 30. The pharmaceutical composition of any of embodiments 22-29, which comprises PEG-monostearate.

[0067] 31. The pharmacal composition of any of embodiments 22-30, which comprises petrolatum.

[0068] 32. The pharmacal composition of any of embodiments 22-31, which is substantially free of water.

[0069] 33. The pharmacal composition of any of embodiments 22-32, which comprises glycerol and / or propylene glycol. 34. The pharmaceutical composition of any of embodiments 22-33, which comprises a cellulosic polymer (e.g., ethylcellulose, e.g., at 1 to 5%).

[0070] 35. A pharmaceutical composition comprising quinine or a salt thereof, wherein the pharmaceutical composition is formulated as a suppository', and wherein the pharmaceutical composition: a) is free of, is substantially free of, or comprises less than 16%, 10%, 5%, 1%, 0.1%, or 0.01% by weight of, purified leaf extract of Neem; b) is free of. is substantially free of, or comprises less than 8%, 4%, 2%, 1%, 0.1%, or 0.01% by weight of, purified saponins; c) is free of, is substantially free of, or comprises less than 8%, 4%, 2%, 1%, 0. 1%, or 0.01% byweight of, purified saponins from Sapindns muckerossi; and / or d) is free of, is substantially free of, or comprises less than 4%, 2%, 1%, 0.1%, or 0.01% by weight of, Mentha citrata oil.

[0071] 36. Tire pharmaceutical composition of embodiment 35, which is free of: a) and b); a) and c); a) and d); b) and c); b) and d); and / or c) and d).

[0072] 37. The pharmaceutical composition of embodiment 35, which is substantially free of: a) and b); a) and c); a) and d); b) and c); b) and d); and / or c) and d). 38. The pharmaceutical composition of embodiment 35, which comprises less than 1%, 0.1%, or 0.01% by weight of each of: a) and b); a) and c); a) and d); b) and c); b) and d); and / or c) and d).

[0073] 39. A method of treating a mucosal epithelial cancer (e.g., vulvar, vaginal, anal, or oropharyngeal malignancy), oral mucosal neoplasia, or anal mucosal neoplasia, cutaneous precancerous and cancerous lesions (e.g., actinic keratosis), a squamous cell cancer, warts (e.g., genital warts), cervical cancer, or HPV infection with HPV 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, or 59, in a subject in need thereof, comprising administering to the subject an effective amount of a pharmaceutical composition comprising quinine or a salt thereof.

[0074] 40. Use of a pharmaceutical composition comprising quinine or a salt thereof in the manufacture of a medicament for treating a mucosal epithelial cancer (e.g., vulvar, vaginal, anal, or oropharyngeal malignancy), oral mucosal neoplasia, or anal mucosal neoplasia, cutaneous precancerous and cancerous lesions (e.g., actinic keratosis), a squamous cell cancer, warts (e.g., genital warts), cervical cancer, or HPV infection with HPV 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, or 59 in a subject in need thereof.

[0075] 41. A pharmaceutical composition comprising quinine or a salt thereof for use in treating a mucosal epithelial cancer (e.g., vulvar, vaginal, anal, or oropharyngeal malignancy), oral mucosal neoplasia, or anal mucosal neoplasia, cutaneous precancerous and cancerous lesions (e.g., actinic keratosis), a squamous cell cancer, warts (e.g., genital warts), cervical cancer, or HPV infection with HPV 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, or 59, in a subject in need thereof.

[0076] 42. A method of treating a mucosal epithelial cancer (e.g., vulvar, vaginal, anal, or oropharyngeal malignancy), an epithelial neoplasia (e.g., cervical dysplasia, genital mucosal neoplasia, oral mucosal neoplasia, oropharyngeal dysplasia, oropharyngeal neoplasia, or anal mucosal neoplasia), cutaneous precancerous and cancerous lesions (e.g., actinic keratosis), a squamous cell cancer, warts (e.g., genital warts), cervical cancer, or HPV infection, in a subject in need thereof, comprising administering to the subject an effective amount of a pharmaceutical composition comprising zinc gluconate. 43. Use of a pharmaceutical composition comprising zinc gluconate in the manufacture of a medicament for treating a mucosal epithelial cancer (e.g., vulvar, vaginal, anal, or oropharyngeal malignancy), an epithelial neoplasia (e.g., cervical dysplasia, genital mucosal neoplasia, oral mucosal neoplasia, oropharyngeal dysplasia, oropharyngeal neoplasia, or anal mucosal neoplasia), cutaneous precancerous and cancerous lesions (e.g., actinic keratosis), a squamous cell cancer, warts (e.g., genital warts), cervical cancer, or HPV infection, in a subject in need thereof.

[0077] 44. A pharmaceutical composition for use in treating a mucosal epithelial cancer (e.g., vulvar, vaginal, anal, or oropharyngeal malignancy), an epithelial neoplasia (e.g., cervical dysplasia, genital mucosal neoplasia, oral mucosal neoplasia, oropharyngeal dysplasia, oropharyngeal neoplasia, or anal mucosal neoplasia), cutaneous precancerous and cancerous lesions (e.g., actinic keratosis), a squamous cell cancer, warts (e.g., genital warts), cervical cancer, or HPV infection, in a subject in need thereof.

[0078] 45. A method of treating a mucosal epithelial cancer (e.g., vulvar, vaginal, anal, or oropharyngeal malignancy), an epithelial neoplasia (e.g., cervical dysplasia, genital mucosal neoplasia, oral mucosal neoplasia, oropharyngeal dysplasia, oropharyngeal neoplasia, or anal mucosal neoplasia), cutaneous precancerous and cancerous lesions (e.g., actinic keratosis), a squamous cell cancer, warts (e.g., genital warts), cervical cancer, or HPV infection, in a subject in need thereof, comprising administering to the subject an effective amount of a pharmaceutical composition comprising zinc or a salt thereof, wherein the method does not comprise administration of or does not comprise administration of a substantial amount of (e.g., the composition is free of, or substantially free of, e.g., comprises less than 1%. 0.1%, or 0.01% by weight of each of): a) Sanguinaria canadensis (bloodroot); or b) bromelain.

[0079] 46. A method of treating a mucosal epithelial cancer (e.g., vulvar, vaginal, anal, or oropharyngeal malignancy), an epithelial neoplasia (e.g., cervical dysplasia, genital mucosal neoplasia, oral mucosal neoplasia, oropharyngeal dysplasia, oropharyngeal neoplasia, or anal mucosal neoplasia), cutaneous precancerous and cancerous lesions (e.g., actinic keratosis), a squamous cell cancer, warts (e.g.. genital warts), cervical cancer, or HPV infection, in a subject in need thereof, comprising administering to the subject an effective amount of a pharmaceutical composition comprising zinc or a salt thereof, wherein the method does not comprising administering an escharotic treatment, e.g., the pharmaceutical composition is not escharotic.

[0080] 47. A pharmaceutical composition comprising EGCG and quinine or a salt thereof. 48. A pharmaceutical composition comprising EGCG and zinc or a salt thereof.

[0081] 49. The pharmaceutical composition of embodiment 48, wherein the EGCG is complexed with the zinc.

[0082] 50. A pharmaceutical composition comprising zinc or a salt thereof and quinine or a salt thereof.

[0083] 51. A pharmaceutical composition comprising EGCG, quinine or a salt thereof, and zinc and a salt thereof.

[0084] 52. A pharmaceutical composition comprising EGCG complexed with sucrose.

[0085] 53. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of. or comprises less than 75%, 50%, 25%, 10%, 1%, 0.1 %, or 0.01 % by weight of, bromelain.

[0086] 54. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of, or comprises less than 75%, 50%, 25%, 10%, 1%, 0.1%, or 0.01% by weight of, Sanguinaria canadensis (bloodroot).

[0087] 55. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is not an escharotic composition.

[0088] 56. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of, or comprises less than 1%, 0.1 %, or 0.01% by weight of Hydrastis, e.g., Hydrastis tincture.

[0089] 57. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of, or comprises less than 1%, 0.1%, or 0.01% by weight of thuja oil.

[0090] 58. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of, or comprises less than 1%, 0.1%, or 0.01% by weight of tea tree oil.

[0091] 59. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of, or comprises less than 1%, 0.1%, or 0.01% by weight of bitter orange oil. 60. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of, or comprises less than 1%, 0.1%, or 0.01% by weight of vitamin A.

[0092] 61 . The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of, or comprises less than 1%, 0.1%, or 0.01% by weight of vitamins.

[0093] 62. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of, or comprises less than 1%, 0.1%, or 0.01% by weight of iron.

[0094] 63. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of, or comprises less than 1%, 0.1%. or 0.01% byweight of ferric sulfate.

[0095] 64. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of. or comprises less than 1%, 0.1%. or 0.01% by weight of ferrous sulfate.

[0096] 65. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the pharmaceutical composition has no more than 10, 9, 8, 7, 6, 5, 4. or 3 active ingredients.

[0097] 66. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the amount of EGCG is 175 to 500 milligrams per suppository.

[0098] 67. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the amount of EGCG is 200 to 400 milligrams per suppository.

[0099] 68. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the amount of EGCG is about 62.5 milligrams per suppository.

[0100] 69. Tire pharmaceutical composition, method, use, or composition for use of any of tire preceding embodiments, wherein the amount of EGCG is about 25-125 or 50-100 milligrams per suppository, e.g., 50 mg or 100 mg. 70. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the amount of quinine or a salt thereof is 40 to 200 milligrams per suppository.

[0101] 71. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the amount of quinine or a salt thereof is 60 to 120 milligrams per suppository.

[0102] 72. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the amount of quinine or a salt thereof is about 7 milligrams per suppository.

[0103] 73. The pharmaceutical composition, method, use, or composition for use of any embodiments 1-71, wherein the amount of quinine or a salt thereof is about 62.5 milligrams per suppository.

[0104] 74. The pharmaceutical composition, method, use, or composition for use of any embodiments 1-71, wherein the amount of quinine or a salt thereof is about 25-125 or 50-100 milligrams per suppository, e.g., 50 mg or 100 mg.

[0105] 75. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the amount of zinc or a salt thereof is about 1 milligram per suppository.

[0106] 76. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the amount of zinc or a salt thereof is about 0.2-0.3 or 0.24 milligram per suppository.

[0107] 77. The pharmaceutical composition, method, use, or composition for use of any embodiments 1-74, wherein the amount of zinc or a salt thereof is 5-20 milligrams per suppository, e.g., 7.5-17.5 milligrams per suppository, e.g., 10-15 milligrams per suppository.

[0108] 78. The pharmaceutical composition, method, use, or composition for use of any of embodiments 1- 74 or 77, wherein the amount of zinc or a salt thereof is about 12.5 milligram per suppository.

[0109] 79. The pharmaceutical composition of any of the preceding embodiments, which further comprises EGCG

[0110] 80. The pharmaceutical composition of any of the preceding embodiments, which further comprises quinine or a salt thereof.

[0111] 81. Tire pharmaceutical composition of any of the preceding embodiments, which further comprises zinc or a salt thereof, e.g., zinc acetate or zinc gluconate. 82. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the EGCG is present at a higher amount by weight than the quinine or salt thereof.

[0112] 83. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the ratio of the EGCG to the quinine or salt thereof is greater than 2, 3, or 4.

[0113] 84. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the EGCG is present at an amount at least 2, 3, 4 times the amount of the quinine or salt thereof.

[0114] 85. The pharmaceutical composition, method, use, or composition for use of any of embodiments 1- 81, wherein the EGCG is present at an equal amount by weight as the quinine or salt thereof.

[0115] 86. The pharmaceutical composition, method, use, or composition for use of any of embodiments 1- 81 or 85, wherein the ratio of the EGCG to the quinine or salt thereof is about 1 .

[0116] 87. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the EGCG is present at a higher amount by weight than the zinc or salt thereof.

[0117] 88. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the ratio of the EGCG to the zinc or salt thereof is greater than 200, 300, or 400.

[0118] 89. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the EGCG is present at an amount at least 200. 300, or 400 times the amount of the zinc or salt thereof.

[0119] 90. The pharmaceutical composition, method, use, or composition for use of any of embodiments 1- 87, wherein the ratio of the EGCG to the zinc or salt thereof is greater than 3, or 4, or 5.

[0120] 91. The pharmaceutical composition, method, use, or composition for use of any of embodiments 1- 87 or 90, wherein the EGCG is present at an amount at least 3, 4, or 5 times the amount of the zinc or salt thereof.

[0121] 92. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the quinine or salt thereof is present at a higher amount by weight than the zinc or salt thereof. 93. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the ratio of the quinine or salt thereof to the zinc or salt thereof is greater than 70, 90, or 110.

[0122] 94. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the quinine or salt thereof is present at an amount at least 70, 90, or 110 times the amount of the zinc or salt thereof.

[0123] 95. The pharmaceutical composition, method, use, or composition for use of any of embodiments 1- 92, wherein the ratio of the quinine or salt thereof to the zinc or salt thereof is greater than 3, or 4, or 5.

[0124] 96. The pharmaceutical composition, method, use, or composition for use of any of embodiments 1- 92 or 95, wherein the quinine or salt thereof is present at an amount at least 3, 4, or 5 times the amount of the zinc or salt thereof.

[0125] 97. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the EGCG is present at a higher amount by weight than the quinine or salt thereof, and the quinine or salt thereof is present at a higher amount by weight than the zinc or salt thereof.

[0126] 98. Tire pharmaceutical composition, method, use, or composition for use of any of tire preceding embodiments, wherein the EGCG is present at an equal amount by weight to the quinine or salt thereof, and the quinine or salt thereof is present at a higher amount by weight than the zinc or salt thereof.

[0127] 99. Tire pharmaceutical composition, method, use, or composition for use of any of tire preceding embodiments, wherein the ratio of the EGCG to quinine or salt thereof to zinc or salt thereof is 300 + / - 20%: 90 + / - 20% : 1 + / - 20%.

[0128] 100. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the ratio of the EGCG to quinine or salt thereof to zinc or salt thereof is 300 + / - 15%: 90 + / - 15% : 1 + / - 15%.

[0129] 101. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the ratio of the EGCG to quinine or salt thereof to zinc or salt thereof is 300 + / - 10%: 90 + / - 10% : 1 + / - 10%. 102. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the ratio of the EGCG to quinine or salt thereof to zinc or salt thereof is 300 + / - 5%: 90 + / - 5% : 1 + / - 5%.

[0130] 103. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the ratio of the EGCG to quinine or salt thereof to zinc or salt thereof is 4 + / - 20%: 4 + / - 20% : 1 + / - 20%.

[0131] 104. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the ratio of the EGCG to quinine or salt thereof to zinc or salt thereof is 4 + / - 15%: 4 + / - 15% : 1 + / - 15%.

[0132] 105. Tire pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the ratio of the EGCG to quinine or salt thereof to zinc or salt thereof is 4 + / - 10%: 4 + / - 10% : 1 + / - 10%.

[0133] 106. Tire pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the ratio of the EGCG to quinine or salt thereof to zinc or salt thereof is 4 + / - 5%: 4 + / - 5% : 1 + / - 5%.

[0134] 107. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the zinc salt is zinc acetate, zinc gluconate or zinc citrate.

[0135] 108. Tire pharmaceutical composition, method, use, or composition for use of the preceding embodiments, wherein the pharmaceutical composition has a pH of less than 7, e.g.. 3-6, e.g., about 4.5.

[0136] 109. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of. or comprises less than 1%. 0.1%. or 0.01% by weight of, green tea extract.

[0137] 110. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of, or comprises less than 1%, 0.1%, or 0.01% by weight of, catechins other than EGCG.

[0138] 111. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of. or comprises less than 1%, 0.1%. or 0.01% by weight of. (-)-epicatechin (EC). 112. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of, or comprises less than 1%, 0.1%, or 0.01% by weight of, (-)-epicatechin-3 -gallate (ECG).

[0139] 113. Tire pharmaceutical composition, method, use, or composition for use of any of tire preceding embodiments, which is free of, is substantially free of. or comprises less than 1%, 0.1%. or 0.01% by weight of (-)-epigallocatechin (EGC).

[0140] 114. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of, or comprises less than 1%, 0.1%, or 0.01% by weight of gallocatechin.

[0141] 115. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of, or comprises less than 1%, 0.1%, or 0.01% by weight of catechin gallate.

[0142] 116. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of. or comprises less than 1%. 0.1%. or 0.01% by weight of gallocatechin gallate.

[0143] 117. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of, or comprises less than 0.5%, 0.25%, or 0.01% by weight of a catechin other than EGCG.

[0144] 118. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, which is free of, is substantially free of. or comprises less than 1%, 0.1%. or 0.01% by weight of theasinensin A.

[0145] 119. The method, use, or pharmaceutical composition for use of any of the preceding embodiments, wherein the amount of zinc or salt thereof in the composition is less than 5%, 2%, or 1% of the total composition by volume.

[0146] 120. Tire pharmaceutical composition, method, use, or composition for use of any of tire preceding embodiments, wherein the pharmaceutical composition is formulated as a suppository.

[0147] 121. The pharmaceutical composition, method, use, or composition for use of embodiment 120, wherein the suppository is formulated for vaginal administration.

[0148] 122. The pharmaceutical composition, method, use, or composition for use of embodiment 120, wherein the suppository is formulated for anal administration. 123. The pharmaceutical composition, method, use, or composition for use of embodiment 120, wherein the pharmaceutical composition is formulated for topical administration.

[0149] 124. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the pharmaceutical composition is fonnulated for oral administration.

[0150] 125. Tire pharmaceutical composition, method, use, or composition for use of any of tire preceding embodiments, wherein the pharmaceutical composition lacks an oxidized form of EGCG, or which comprises the oxidized form of EGCG at a level less than 50%, 40%, 30%, 20%, 10%, 5%, 3%. 2%, or 1% of the EGCG.

[0151] 126. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the pharmaceutical composition comprises epigallocatechin gallate (EGCG), in a dosage form comprising 25-75 mg, 30-70 mg, 40-60 mg, or 45-55 mg, e.g., 50mg, or 225-500 mg of the EGCG.

[0152] 127. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the pharmaceutical composition lacks an antioxidant other than EGCG, or comprises an antioxidant otherthan EGCG at a level of less than 5, 4, 3, 2, or 1 mg per dosage form (e.g., less than 2.5, 2, 1.5, 1, or 0.5 mg / ml).

[0153] 128. Tire pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the pharmaceutical composition lacks a reducing agent other than EGCG, or comprises a reducing agent other than EGCG at a level of less than 5, 4, 3, 2, or 1 mg per dosage form (e.g., less than 2.5, 2, 1.5, 1, or 0.5 mg / ml).

[0154] 129. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the pharmaceutical composition: a) is free of, is substantially free of, or comprises less than 16%, 10%, 5%, 1%, 0.1%, or 0.01% by weight of, purified leaf extract of Neem; b) is free of, is substantially free of, or comprises less than 8%, 4%, 2%, 1%, 0.1%, or 0.01% by weight of, purified saponins; c) is free of, is substantially free of, or comprises less than 8%, 4%, 2%. 1%, 0. 1%, or 0.01% byweight of. purified saponins from Sapindus muckerossi; and / or d) is free of. is substantially free of. or comprises less than 4%, 2%. 1%, 0.1%, or 0.01% by weight of, Mentha citrata oil. 130. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the EGCG is complexed with the zinc.

[0155] 131. The pharmaceutical composition, method, use, or composition for use of any of the preceding embodiments, wherein the EGCG is complexed with the sucrose.

[0156] 132. A pharmaceutical composition comprising: a first poly-ethylene glycol (PEG) having an average molecular weight between 300-500 g / mol (e.g., PEG400, e.g., PEG400 monostearate). a second PEG having an average molecular weight between 1000-2000 g / mol, (e.g., PEG1500); and a third PEG having an average molecular weight between 3000-5000 g / mol, (e.g., PEG 3350 or PEG4000).

[0157] 133. Tire pharmaceutical composition of embodiment 132, wherein the first PEG has an average molecular weight between 325-475 g / mol, e.g., 350-450 g / mol, e.g., 375-425 g / mol.

[0158] 134. The pharmaceutical composition of embodiment 132 or 133, wherein the second PEG has an average molecular weight between 1200-1800 g / mol, e.g.. 1400-1600 g / mol.

[0159] 135. The pharmaceutical composition of any of embodiments 132-134, wherein the third PEG has an average molecular weight between 3250-4750 g / mol, e.g., 3500-4500 g / mol, e.g., 3750-4250 g / mol.

[0160] 136. The pharmaceutical composition of any of embodiments 132-135, wherein the ratio of the first PEG : second PEG : third PEG is 1 ± 20% : 2 ± 20% : 2 ± 20%, e.g., wherein the ratio is 1 ± 10% : 2 ± 10% : 2 ± 10%.

[0161] 137. The pharmaceutical composition of any of embodiments 132-136, wherein the ratio of the first PEG : second PEG is 1 ± 20% : 2 ± 20%, e.g., wherein the ratio is 1 ± 10% : 2 ± 10%.

[0162] 138. The pharmaceutical composition of any of embodiments 132-137, wherein the ratio of the second PEG : third PEG is 2 ± 20% : 2 ± 20%, e.g., wherein the ratio is 2 ± 10% : 2 ± 10%.

[0163] 139. The pharmaceutical composition of any of embodiments 132-138, wherein the ratio of the first PEG : third PEG is 1 ± 20% : 2 ± 20%, e.g., wherein the ratio is 1 ± 10% : 2 ± 10%.

[0164] 140. A pharmaceutical composition comprising: a first poly-ethylene glycol (PEG) having an average molecular weight between 1000-2000 g / mol, (e.g., PEG1500); and a second PEG having an average molecular weight between 3000-5000 g / mol, (e.g., PEG 3350 or PEG4000).

[0165] 141. Tire pharmaceutical composition of embodiment 140, wherein the first PEG has an average molecular weight between 1200-1800 g / mol, e.g., 1400-1600.

[0166] 142. The pharmaceutical composition of embodiment 140 or 141, wherein the second PEG has an average molecular weight between 3250-4750 g / mol, e.g., 3500-4500 g / mol, e.g., 3750-4250 g / mol.

[0167] 143. The pharmaceutical composition of any of embodiments 140-142, wherein the ratio of the first PEG : second PEG is 2.3 ± 20% : 1 ± 20%, e.g., wherein the ratio is 2.3 ± 10% : 1 ± 10%.

[0168] 144. A pharmaceutical composition comprising: one or more PEGs (e.g., a PEG400, a PEG1500, and a PEG4000); and a hard fat comprising two or more of (e.g., all of): i) a mono-glyceride ester of a fatty acid: ii) a di-glyceride ester of a fatty acid; and iii) a triglyceride ester of a fatty acid; wherein optionally the fatty acids have a carbon chain length of CIO to Cl 8.

[0169] 145. A pharmaceutical composition comprising: one or more PEGs (e.g., a PEG400, a PEG1500, and a PEG4000); and a hard fat comprising two or more of (e.g., all of): i) a mono-glyceride ester of a fatty acid; ii) a di-glyceride ester of a fatty acid; iii) a triglyceride ester of a fatty acid; iv) one or more PEG stearates, e.g., one or more PEG monostearates comprising a PEG having a molecular weight in the range of 200-2000, e.g., one or more PEG monostearates comprising a PEG having a molecular weight in the range of 300-1500; and wherein optionally the fatty acids have a carbon chain length of CIO to Cl 8. 146. The pharmaceutical composition of embodiments 144 and 145 wherein the hard fat comprises saturated fatty acids, e.g., glycerides, e.g., C12-C18 glycerides, e.g., C12-C18 mono-, di-, and triglycerides, e.g., Gattefosse Ovucire WL, or Tefose®, e.g., Tefose® 1500.

[0170] 147. Tire pharmaceutical composition of embodiment 144-146, wherein the triglyceride ester is more abundant than the monogylceride ester.

[0171] 148. The pharmaceutical composition of any of embodiments 144-147, wherein the triglyceride ester is more abundant than the digylceride ester.

[0172] 149. The pharmaceutical composition of any of embodiments 144-148, which further comprises an emulsifier, e.g., an ethoxylated fatty alcohol, e.g., ceteth-20 and / or steareth-20.

[0173] 150. The pharmaceutical composition of any of embodiments 144-149, wherein the ratio of the one or more PEGs to the hard fat is 25 ± 20% : 8 ± 20%, e.g., 25 ± 10% : 8 ± 10%.

[0174] 151. A pharmaceutical composition comprising: one or more PEG (e.g., a PEG400, a PEG 1500, a PEG 3350, and / or a PEG4000); and hydroxypropyl methylcellulose (HPMC) (e.g., HPMC KI OOM).

[0175] 152. The pharmaceutical composition of embodiment 151, wherein tire EIPMC has a molecular weight of 500,000-1,500,000 daltons, e.g., about 1,000.000 daltons.

[0176] 153. The pharmaceutical composition of embodiment 151 or 152, wherein the HPMC has a molecular weight of at least 500,000, at least 600,000, at least 700,000, at least 800,000, at least 900,000, or at least 1,000,000 daltons.

[0177] 154. Tire pharmaceutical composition of any of embodiments 151-153, wherein the HPMC has a viscosity of 50,000 - 150,000 mPas (millipascal seconds), e.g., 75,000 - 125,000 mPas, e.g., about 100,000 mPas.

[0178] 155. The pharmaceutical composition of any of embodiments 151-154, wherein the HPMC has a viscosity of at least 50,000 mPas, at least 60,000 mPas, at least 70,000 mPas, at least 80,000 mPas, at least 90,000 mPas, or at least 100,000 mPas.

[0179] 156. The pharmaceutical composition of any of embodiments 151-155, wherein the viscosity of the HPMC is measured at 2% concentration in water.

[0180] 157. Tire pharmaceutical composition of any of embodiments 151-156, wherein the HPMC is a K100M grade HPMC. 158. The pharmaceutical composition of any of embodiments 151-157, wherein the HPMC has a particle size of 100 to 300 pm, e.g., 150 to 250 pm, e.g., 170 to 250 pm.

[0181] 159. The pharmaceutical composition of any of embodiments 151-158, wherein the HPMC has a methoxyl content of 10-35%, e.g., 15-30%, e.g., 20-25%, e.g., 20-24%.

[0182] 160. The pharmaceutical composition of any of embodiments 151-159, wherein the HPMC has a hydroxypropyl content of 1-18%, e.g.. 4-15%. e g., 7-12%.

[0183] 161. The pharmaceutical composition of any of embodiments 151-160, wherein the HPMC has a use level of 0.05-5% polymer, e.g., 0.1-3% polymer, e.g., 0.2-2% polymer.

[0184] 162. The pharmaceutical composition of any of embodiments 151-161, wherein the ratio of the one or more PEGs to the HPMC is 25 ± 20% : 3 ± 20%, e.g., 25 ± 10% : 3 ± 10%.

[0185] 163. The pharmaceutical composition of any of embodiments 151-162, which further comprises one or more active ingredients.

[0186] 164. The pharmaceutical composition of any of embodiments 151-163, which further comprises PEGoxol-7 Stearate.

[0187] 165. The pharmaceutical composition of any of embodiments 151-163, which further comprises PEG 400 stearate.

[0188] 166. The pharmaceutical composition of any of embodiments 151-165, which further comprises hydroxyethylcellulose (HEC).

[0189] 167. A pharmaceutical composition comprising: epigallocatechin gallate (EGCG) or a salt thereof; and sucrose or a salt thereof, wherein the pharmacal composition is formulated as a suppository.

[0190] 168. The pharmaceutical composition of embodiment 167, which further comprises citric acid or a salt thereof (e.g., citric acid monohydrate).

[0191] 169. A pharmaceutical composition comprising: a first poly-ethylene glycol (PEG) having an average molecular weight between 300-500 g / mol (e.g., PEG400), a second PEG having an average molecular weight between 1000-2000 g / mol, (e.g., PEG1500); and a third PEG having an average molecular weight between 3000-5000 g / mol, (e.g., PEG4000).

[0192] 170. Tire pharmaceutical composition of embodiment 169, wherein the first PEG has an average molecular weight between 325-475 g / mol, e.g., 350-450 g / mol, e.g., 375-425 g / mol.

[0193] 171. The pharmaceutical composition of embodiment 169 or 170, wherein the second PEG has an average molecular weight between 1200-1800 g / mol, e.g.. 1400-1600 g / mol.

[0194] 172. The pharmaceutical composition of any of embodiments 169-171, wherein the third PEG has an average molecular weight between 3250-4750 g / mol, e.g., 3500-4500 g / mol, e.g., 3750-4250 g / mol.

[0195] 173. The pharmaceutical composition of any of embodiments 169- 172, wherein the ratio of the first PEG : second PEG : third PEG is 1 ± 20% : 2 ± 20% : 2 ± 20%, e.g., wherein the ratio is 1 ± 10% : 2 ± 10% : 2 ± 10%.

[0196] 174. Tire pharmaceutical composition of any of embodiments 169-173, wherein the ratio of the first PEG : second PEG is 1 ± 20% : 2 ± 20%, e.g., wherein the ratio is 1 ± 10% : 2 ± 10%.

[0197] 175. The pharmaceutical composition of any of embodiments 169-174, wherein the ratio of the second PEG : third PEG is 2 ± 20% : 2 ± 20%, e.g., wherein the ratio is 2 ± 10% : 2 ± 10%.

[0198] 176. The pharmaceutical composition of any of embodiments 169-175, wherein the ratio of the first PEG : third PEG is 1 ± 20% : 2 ± 20%, e.g., wherein the ratio is 1 ± 10% : 2 ± 10%.

[0199] 177. Tire pharmaceutical composition of any of embodiments 169-176, which further comprises a hard fat.

[0200] 178. The pharmaceutical composition of embodiment 177, wherein the hard fat comprises two or more of (e.g., all of): i) a mono-glyceride ester of a fatty acid; ii) a di-glyceride ester of a fatty acid; and iii) a triglyceride ester of a fatty acid.

[0201] 179. The pharmaceutical composition of embodiment 177 wherein the hard fat comprises hard fat Polyoxyl 20 cetostearyl ether, e.g., Gattefosse Ovucire WL.

[0202] 180. The pharmaceutical composition of embodiment 177 wherein the hard fat comprises Tefose®, e.g., Tefose® 1500.

[0203] 181. Tire pharmaceutical composition of embodiment 178 or 180, wherein the triglyceride ester is more abundant than the monogylceride ester. 182. The pharmaceutical composition of any of embodiments 178-181, wherein the triglyceride ester is more abundant than the digylceride ester.

[0204] 183. The pharmaceutical composition of any of embodiments 151-182, which further comprises an emulsifier, e.g., an ethoxylated fatty alcohol.

[0205] 184. The pharmaceutical composition of any of embodiments 132-183, which further comprises a surfactant, wherein optionally the surfactant is HPMC.

[0206] 185. The pharmaceutical composition of any of embodiments 132-184, which further comprises an emulsifier, wherein optionally the emulsifier is HPMC.

[0207] 186. The pharmaceutical composition of any of embodiments 132-185, which further comprises a mucoadhesive agent, wherein optionally the mucoadhesive agent is HPMC.

[0208] 187. The pharmaceutical composition of any of embodiments 151-186, wherein the HPMC has a molecular weight of 500,000-1,500,000 daltons, e.g., about 1,000,000 daltons.

[0209] 188. The pharmaceutical composition of any of embodiments 151-187, wherein the HPMC has a molecular weight of at least 500,000, at least 600,000. at least 700,000, at least 800,000. at least 900,000, or at least 1.000.000 daltons.

[0210] 189. The pharmaceutical composition of any of embodiments 151-188, wherein the HPMC has a viscosity of 50,000 - 150,000 mPas, e.g., 75,000 - 125,000 mPas, e.g., about 100,000 mPas.

[0211] 190. The pharmaceutical composition of any of embodiments 151-189, wherein the HPMC has a viscosity of at least 50,000 mPas, at least 60,000 mPas, at least 70,000 mPas, at least 80,000 mPas, at least 90,000 mPas, or at least 100,000 mPas.

[0212] 191. Hie pharmaceutical composition of any of embodiments 151-190, wherein the viscosity of the HPMC is measured at 2% concentration in water.

[0213] 192. The pharmaceutical composition of any of embodiments 151-191. wherein the HPMC is a KI OOM grade HPMC.

[0214] 193. The pharmaceutical composition of any of embodiments 151-192, wherein the HPMC has a particle size of 100 to 300 pm, e.g., 150 to 250 pm, e.g., 170 to 250 pm.

[0215] 194. The pharmaceutical composition of any of embodiments 151-193, wherein the HPMC has a methoxyl content of 10-35%, e.g., 15-30%, e.g., 20-25%, e.g., 20-24%. 195. The pharmaceutical composition of any of embodiments 151-194, wherein the HPMC has a hydroxypropyl content of 1-18%, e.g., 4-15%, e.g., 7-12%.

[0216] 196. The pharmaceutical composition of any of embodiments 151-195, wherein the HPMC has a use level of 0.05-5% polymer, e.g., 0.1-3% polymer, e.g., 0.2-2% polymer.

[0217] 197. Tire pharmaceutical composition of any of embodiments 132-196, which further comprises a colorant, e.g., wherein the colorant comprises astaxanthin or a salt thereof, or canthaxanthin or a salt thereof.

[0218] 198. The pharmaceutical composition of any of embodiments 132-197, which is free of, is substantially free of, or comprises less than 1%, 0.1 %, or 0.01 % by weight of, a poly(acrylic acid), e g., carbopol .

[0219] 199. Tire pharmaceutical composition of any of embodiments 132-198, which is free of, is substantially free of, or comprises less than 1%, 0.1%, or 0.01% by weight of, a polysorbate, e.g., a Tween polysorbate.

[0220] 200. The pharmaceutical composition of any of embodiments 132-199, which further comprises one or more stabilizing agents, wherein optionally one of the one or more stabilizing agents is citric acid or a salt thereof (e.g., citric acid monohydrate).

[0221] 201. The pharmaceutical composition of any of embodiments 132-200, wherein one of the one or more stabilizing agents is sucrose or a salt thereof.

[0222] 202. Tire pharmaceutical composition of any of embodiments 132-201, which further comprises citric acid or a salt thereof, wherein optionally the citric acid or salt thereof is present at a level of 2-20 mg per suppository, e.g., 5-15 mg per suppository, e.g., about 11 mg per suppository.

[0223] 203. The pharmaceutical composition of any of embodiments 132-201. which further comprises citric acid or a salt thereof, wherein optionally the citric acid or salt thereof is present at a level of 0.01-1 mg per suppository, e.g., 0.05-0.5 mg per suppository, e.g., 0. 1-0.2 mg per suppository, e.g., about 0.24 mg per suppository.

[0224] 204. Tire pharmaceutical composition of any of embodiments 132-203, which further comprises sucrose or a salt thereof.

[0225] 205. The pharmaceutical composition of embodiment 204, wherein the sucrose or salt thereof is present at a level of 40-70 mg. e.g., about 55 mg or 50 mg per suppository. 206. The pharmaceutical composition of embodiment 204, wherein the sucrose or salt thereof is present at a level of 0.25-1.25 times the amount of EGCG in grams, e.g., 0.5-1 times the amount of EGCG in grams, e.g., about 0.75 or 1 times the amount of EGCG in grams.

[0226] 207. The pharmaceutical composition of embodiment 204, wherein the sucrose or salt thereof is present at 0.5 to 2.0 times the molar amount of EGCG.

[0227] 208. Tire pharmaceutical composition of any of embodiments 204 or 207, wherein the sucrose or salt thereof is present at a ratio of 1 mol sucrose to 1 mol EGCG.

[0228] 209. A pharmaceutical composition comprising: a first poly-ethylene glycol (PEG) having an average molecular weight between 300-500 g / mol (e.g., PEG400), a second PEG having an average molecular weight between 2850-3350 g / mol, (e.g., PEG3350); a hard fat comprising:

[0229] (a) i) a mono-glyccridc ester of a fatty acid; ii) a di-glyccridc ester of a fatty acid; and iii) a triglyceride ester of a fatty acid; and / or

[0230] (b) one or more PEG stearates, e.g., one or more PEG monostearates comprising a PEG having a molecular weight in the range of 200-2000, e.g., one or more PEG monostearates comprising a PEG having a molecular weight in the range of 300-1500; hydroxypropyl methylcellulose (EIPMC); and citric acid or a salt thereof (e.g., citric acid monohydrate).

[0231] 210. Tire pharmaceutical composition of embodiment 209, wherein the ratio of the one or more PEGs to the citric acid or salt thereof is 25 ± 20% : 0.2 ± 20%, e.g., 25 ± 10% : 0.2 ± 10%.

[0232] 211. Tire pharmacal composition of embodiment 209 or 210, wherein the ratio of tire hard fat to the HPMC is 8 ± 20% : 3 ± 20%, e.g., 8 ± 10% : 3 ± 10%.

[0233] 212. The pharmaceutical composition of any of embodiments 209-211, wherein the ratio of the HPMC to the citric acid is 3 ± 20% : 0.2 ± 20%, e.g., 3 ± 10% : 0.2 ± 10%.

[0234] 213. The pharmaceutical composition of any of embodiments 209-212, which further comprises sucrose or salt thereof. 214. The pharmaceutical composition of embodiment 213, wherein the ratio of one or more PEGs to the sucrose or salt thereof is 25 ± 20% : 1 ± 20%, e.g., 25 ± 10% : 1 ± 10%.

[0235] 215. The pharmaceutical composition of embodiment 213 or 214, wherein the ratio of the hard fat to the sucrose or salt thereof is 8± 20% : 1 ± 20%, e.g., 8 ± 10% : 1 ± 10%.

[0236] 216. The pharmaceutical composition of any of embodiments 213-215, wherein the ratio of the HPMC to the sucrose or salt thereof is 3± 20% : 1 ± 20%, e.g., 3 ± 10% : 1 ± 10%.

[0237] 217. The pharmacal composition of any of embodiments 213-216, wherein the ratio of the citric acid or salt thereof to the sucrose or salt thereof is 0.2± 20% : 1 ± 20%, e.g., 0.2 ± 10% : 1 ± 10%.

[0238] 218. The pharmaceutical composition of any of embodiments 213-217, wherein the ratio of one or more PEGs to the EIPMC to the citric acid or salt thereof is 25± 20% : 3 ± 20% : 0.2 ± 20%, e.g., 25 ± 10% : 3 ± 10% : 0.2 ± 10%.

[0239] 219. The pharmaceutical composition of any of embodiments 213-218, wherein the ratio of one or more PEGs to the HPMC to the citric acid or salt thereof to the sucrose or salt thereof is 25± 20% : 3 ± 20% : 0.2 ± 20% : 1 ± 20%. e.g., 25 ± 10% : 3 ± 10% : 0.2 ± 10% : 1 ± 10%.

[0240] 220. The pharmaceutical composition of any of embodiments 209-219, wherein the first PEG has an average molecular weight between 325-475 g / mol, e.g., 350-450 g / mol, e.g., 375-425 g / moL

[0241] 221 . The pharmaceutical composition of any of embodiments 209-220, wherein the second PEG has an average molecular weight between 1200-1800 g / mol, e.g., 1400-1600 g / mol.

[0242] 222. Tire pharmaceutical composition of any of embodiments 209-221, wherein the third PEG has an average molecular weight between 3250-4750 g / mol, e.g., 3500-4500 g / mol, e.g., 3750-4250 g / mol.

[0243] 223. The pharmaceutical composition of any of embodiments 209-222, wherein the ratio of the first PEG : second PEG : third PEG is 1 ± 20% : 2 ± 20% : 2 ± 20%, e.g., wherein the ratio is 1 ± 10% : 2 ± 10% : 2 ± 10%.

[0244] 224. The pharmaceutical composition of any of embodiments 209-223, wherein the ratio of the first PEG : second PEG is 1 ± 20% : 2 ± 20%, e.g., wherein the ratio is 1 ± 10% : 2 ± 10%.

[0245] 225. The pharmaceutical composition of any of embodiments 209-224, wherein the ratio of the second PEG : third PEG is 2 ± 20% : 2 ± 20%, e.g., wherein the ratio is 2 ± 10% : 2 ± 10%. 226. The pharmaceutical composition of any of embodiments 209-225, wherein the ratio of the first PEG : third PEG is 1 ± 20% : 2 ± 20%, e.g., wherein the ratio is 1 ± 10% : 2 ± 10%.

[0246] 227. Tire pharmaceutical composition of any of embodiments 209-226, wherein the hard fat comprises saturated fatty acids, e.g., glycerides, e.g., C12-C18 glycerides, e.g., C12-C18 mono-, di-, and tri- glycerides, e.g., Gattefosse Ovucire WL, or Tefose®, e.g., Tefose® 1500.

[0247] 228. The pharmaceutical composition of any of embodiments 209-227, wherein the triglyceride ester is more abundant than the monogylceride ester.

[0248] 229. The pharmaceutical composition of any of embodiments 209-228, wherein the triglyceride ester is more abundant than the digylceride ester.

[0249] 230. Tire pharmaceutical composition of any of embodiments 209-229, which further comprises an emulsifier, e.g., an ethoxylated fatty alcohol, e.g., ceteth-20 and / or steareth-20.

[0250] 231. Tire pharmaceutical composition of any of embodiments 209-230, wherein the HPMC has a molecular weight of 500,000-1,500,000 daltons, e.g., about 1,000,000 daltons.

[0251] 232. The pharmaceutical composition of any of embodiments 209-231, wherein the HPMC has a molecular weight of at least 500.000, at least 600,000. at least 700,000, at least 800,000. at least 900,000, or at least 1,000,000 daltons.

[0252] 233. The pharmaceutical composition of any of embodiments 209-232, wherein the HPMC has a viscosity of 50,000 - 150,000 mPas, e.g., 75,000 - 125,000 mPas, e.g., about 100,000 mPas.

[0253] 234. Tire pharmaceutical composition of any of embodiments 209-233, wherein the HPMC has a viscosity of at least 50,000 mPas, at least 60,000 mPas, at least 70,000 mPas, at least 80.000 mPas, at least 90,000 mPas, or at least 100,000 mPas.

[0254] 235. The pharmaceutical composition of any of embodiments 209-234, wherein the viscosity of the HPMC is measured at 2% concentration in water.

[0255] 236. The pharmaceutical composition of any of embodiments 209-235, wherein the HPMC is a K100M grade HPMC.

[0256] 237. The pharmaceutical composition of any of embodiments 151-236, wherein the HPMC has a particle size of 100 to 300 pm, e.g., 150 to 250 pm, e.g., 170 to 250 pm.

[0257] 238. Hie pharmaceutical composition of any of embodiments 151-237, wherein the HPMC has a methoxyl content of 10-35%, e.g., 15-30%, e.g.. 20-25%, e.g.. 20-24%. 239. The pharmaceutical composition of any of embodiments 151-238, wherein the HPMC has a hydroxypropyl content of 1-18%, e.g., 4-15%, e.g., 7-12%.

[0258] 240. The pharmaceutical composition of any of embodiments 151-239, wherein the HPMC has a use level of 0.05-5% polymer, e.g., 0.1-3% polymer, e.g., 0.2-2% polymer.

[0259] 241. The pharmaceutical composition of any of embodiments 132-240, wherein the suppository comprises 0.01, 0.02, 0.03, 0.04, 0.05. 0.06, 0.07, 0.08, 0.09, or 0.1 g of EGCG or a salt thereof.

[0260] 242. The pharmacal composition of any of embodiments 132-241, wherein the suppository compnses 0.01-0.1, 0.03-0.08, or 0.05-0.07 g of EGCG.

[0261] 243. The pharmaceutical composition of any of embodiments 132-242, wherein the suppository comprises 0.04 to 0.2, or 0.06 to 0.12 g of quinine or a salt thereof.

[0262] 244. The pharmaceutical composition of any of embodiments 132-242, wherein the suppository comprises about 62.5 mg of quinine or a salt thereof.

[0263] 245. The pharmaceutical composition of any of embodiments 132-242, wherein the suppository comprises about 50 mg of quinine or a salt thereof.

[0264] 246. The pharmacal composition of any of embodiments 132-245, wherein the suppository comprises 5-20, 7.5-17.5, or 10-15 mg of zinc or a salt thereof.

[0265] 247. The pharmaceutical composition of any of embodiments 132-246, wherein the suppository' comprises about 12.5 mg or 5mg of zinc or a salt thereof.

[0266] 248. The pharmaceutical composition of any of embodiments 132-247, wherein the suppository comprises 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg of citric acid or a salt thereof (e.g., citric acid monohydrate).

[0267] 249. The pharmacal composition of any of embodiments 132-248, wherein the suppository comprises 5-20. 7-18, 9-16, or 11-14 mg of citric acid or a salt thereof (e.g.. citric acid monohydrate).

[0268] 250. The pharmaceutical composition of any of embodiments 132-249, wherein the suppository comprises 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, or 0.1 g of sucrose or a salt thereof.

[0269] 251. The pharmaceutical composition of any of embodiments 132-250, wherein the suppository comprises 0.01-0.1, 0.03-0.08, or 0.05-0.07 g of sucrose or a salt thereof.

[0270] 252. The pharmaceutical composition of any of embodiments 132-251, wherein the suppository comprises 0.05, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5. or 0.55 g of PEG400. 253. The pharmaceutical composition of any of embodiments 132-252, wherein the suppository comprises 0.05-0.55, 0.15-0.45, or 0.25-0.35 g of PEG400.

[0271] 254. The pharmaceutical composition of any of embodiments 132-253, wherein the suppository comprises 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1, 1.05, 1.1, 1.15, or 1.2 g ofPEG1500.

[0272] 255. The pharmaceutical composition of any of embodiments 132-254, wherein the suppository comprises 0.5-1.2. 0.8-1.1, or 0.95-1.05 g of PEG1500.

[0273] 256. The pharmacal composition of any of embodiments 132-255, wherein the suppository comprises 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, or 0.9 g of PEG4000.

[0274] 257. The pharmaceutical composition of any of embodiments 132-256, wherein the suppository comprises 0.2-0.9, 0.3-0.8, 0.4-0.7, or 0.45-0.55 g of PEG4000.

[0275] 258. The pharmaceutical composition of any of embodiments 132-257, wherein the suppository comprises 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, or 0.85 g of ovucire.

[0276] 259. The pharmaceutical composition of any of embodiments 132-258, wherein the suppository comprises 0.25-0.85, 0.35-0.75, or 0.45-0.65 g of ovucire.

[0277] 260. A pharmaceutical composition comprising:

[0278] 1-4%. e.g., 1.5-3.5%, e.g.. 2-3%, e.g.. 2.5% w / w quinine;

[0279] 1-4%, e.g., 1.5-3.5%, e.g., 2-3%, e.g., 2.5% w / w EGCG;

[0280] 8-18%, e.g., 10-16%, e.g., 12-14%, e.g., 12.9% w / w PEG 400:

[0281] 15-35%, e.g., 20-30%, e.g., 22.5-27.5%, e.g., 25.3% w / w PEG 1500;

[0282] 15-35%, e.g., 20-30%, e.g., 22.5-27.5%, e.g., 25.3% w / w PEG 3350 or PEG 4000;

[0283] 12.5-27.5%, e.g., 15-25%, e.g., 17.5-22.5%, e.g., 20.3% w / w Ovucire WL3264;

[0284] 0.1-1%, e.g., 0.2-0.8%, e.g., 0.3-0.7%, e.g., 0.4-0.6%, e.g., 0.5% w / w citric acid (e.g., citric acid monohydrate);

[0285] 1-3%, e.g., 1.5-2.5%, e.g.. 2-3%, e.g., 2.5% w / w sucrose;

[0286] 0.1-1%, e.g., 0.2-0.9%, e.g.. 0.4-0.7%, e.g., 0.6% w / w zinc gluconate: and

[0287] 2.5-12.5%, e.g., 5-10%, e.g., 6-9%, e.g.. 7.6% w / w HPMC. 261. A pharmaceutical composition comprising:

[0288] 1-4%, e.g., 1 .5-3.5%, e.g., 2-3%, e.g.,2.5% w / w quinine:

[0289] 1-4%, e.g., 1.5-3.5%, e.g., 2-3%, e.g.,2.5% w / w EGCG;

[0290] 8-18%, e.g., 10-16%, e.g., 12-14%, e.g., 13.21% w / w PEG 400;

[0291] 15-35%, e.g., 20-30%, e.g., 22.5-27.5%, e.g., 25.3% w / w PEG 1500;

[0292] 15-35%, e.g., 20-30%, e.g., 22.5-27.5%, e.g., 25.3% w / w PEG 3350 or PEG 4000;

[0293] 12.5-27.5%, e.g., 15-25%. e.g., 17.5-22.5%, e.g., 20 % w / w PEGoxol-7 Stearate;

[0294] 0. 1-1%, e.g., 0.2-0.8%, e.g., 0.3-0.7%, e.g., 0.4-0.6%, e.g., 0.49% w / w citric acid (e.g., citric acid monohydrate);

[0295] 1-3%. e.g., 1.5-2.5%, e.g.. 2-3%, e.g.. 2.5% w / w sucrose;

[0296] 0.1-1%, e.g., 0.2-0.9%, e.g., 0.4-0.7%, e.g., 0.6% w / w zinc gluconate; and

[0297] 2.5-12.5%, e.g., 5-10%, e.g., 6-9%, e.g., 7.6% w / w HPMC.

[0298] 262. A pharmaceutical composition comprising:

[0299] 1-4%, e.g., 1.5-3.5%, e.g., 2-3%, e.g., 2.5% w / w quinine;

[0300] 1-4%, e.g., 1.5-3.5%, e.g., 2-3%, e.g., 2.5% w / w EGCG;

[0301] 15-35%, e.g., 20-30%, e.g., 22.5-27.5%, e.g., 24.81% w / w PEG 400;

[0302] 25-55%, e.g., 30-50%, e.g., 35-45%, e.g., 40% w / w PEG 3350 or PEG 4000;

[0303] 12.5-27.5%. e.g., 15-25%. e.g., 17.5-22.5%. e.g., 20% w / w PEGoxol-7 Stearate;

[0304] 0.1-1%, e.g., 0.2-0.8%, e.g., 0.3-0.7%, e.g., 0.4-0.6%, e.g., 0.49% w / w citric acid (e.g., citric acid monohydrate);

[0305] 1-3%. e.g., 1.5-2.5%, e.g.. 2-3%, e.g.. 2.5% w / w sucrose;

[0306] 0.1-1%, e.g., 0.2-0.9%, e.g., 0.4-0.7%, e.g., 0.6% w / w zinc gluconate; and

[0307] 2.5-12.5%, e.g., 5-10%, e.g., 6-9%, e.g., 6.6% w / w HPMC.

[0308] 263. A pharmaceutical composition comprising:

[0309] 1-4%, e.g., 1.5-3.5%, e.g., 2-3%, e.g.,2.5% w / w quinine; 1-4%, e.g., 1.5-3.5%, e.g., 2-3%, e.g.,2.5% w / w EGCG;

[0310] 15-35%, e.g., 20-30%, e.g., 22.5-27.5%, e.g., 24.81% w / w PEG 400;

[0311] 25-55%, e.g., 30-50%, e.g., 35-45%, e.g., 40% w / w PEG 3350 or PEG 4000;

[0312] 12.5-27.5%, e.g., 15-25%, e.g., 17.5-22.5%, e.g., 20% w / w glycerol monostearate;

[0313] 0.1-1%, e.g., 0.2-0.8%, e.g., 0.3-0.7%, e.g., 0.4-0.6%, e.g., 0.49% w / w citric acid (e.g., citric acid monohydrate);

[0314] 1-3%, e.g., 1.5-2.5%, e.g.. 2-3%, e.g., 2.5% w / w sucrose;

[0315] 0.1-1%, e.g., 0.2-0.9%, e.g.. 0.4-0.7%, e.g., 0.6% w / w zinc gluconate; and

[0316] 2.5-12.5%, e.g., 5-10%, e.g., 6-9%, e.g.. 6.6% w / w HPMC.

[0317] 264. A pharmaceutical composition comprising:

[0318] 1-4%, e.g., 1.5-3.5%, e.g., 2-3%, e.g.,2.5% w / w quinine;

[0319] 1-4%, e.g., 1.5-3.5%, e.g., 2-3%, e.g.,2.5% w / w EGCG;

[0320] 8-18%, e.g., 10-16%, e.g., 12-14%, e.g., 12.9% w / w PEG 400;

[0321] 15-35%, e.g., 20-30%, e.g., 22.5-27.5%, e.g., 25.3 % w / w PEG 1500;

[0322] 15-35%, e.g., 20-30%, e.g., 22.5-27.5%, e.g., 25.3 % w / w PEG 3350 or PEG 4000;

[0323] 12.5-27.5%, e.g., 15-25%, e.g., 17.5-22.5%, e.g., 20.3 % w / w Ovucire WL3264;

[0324] 0.1-1%, e.g., 0.2-0.8%, e.g., 0.3-0.7%, e.g., 0.4-0.6%, e.g., 0.5% w / w citric acid (e.g., citric acid monohydrate);

[0325] 1-3%, e.g., 1.5-2. %, e.g.. 2-3%, e.g., 2.5% w / w sucrose;

[0326] 0.1-1%, e.g., 0.2-0.9%, e.g.. 0.4-0.7%, e.g., 0.6% w / w zinc gluconate; and

[0327] 2.5-12.5%, e.g.. 5-10%. e.g., 6-9%, e.g.. 7.6% w / w hydroxyethylcellulose (HEC).

[0328] 265. A pharmaceutical composition comprising:

[0329] 1-4%, e.g., 1.5-3.5%, e.g., 2-3%, e.g., 2.5% w / w quinine;

[0330] 1-4%, e.g., 1.5-3.5%, e.g., 2-3%, e.g., 2.5% w / w EGCG;

[0331] 15-35%, e.g., 20-30%, e.g., 22.5-27.5%, e.g., 24.81% w / w PEG 400; 25-55%, e.g., 30-50%, e.g., 35-45%, e.g., 40% w / w PEG 3350;

[0332] 12.5-27.5%, e.g., 15-25%, e.g., 17.5-22.5%, e.g., 20% w / w Ovucire WL3264;

[0333] 0.1-1%, e.g., 0.2-0.8%, e.g., 0.3-0.7%, e.g., 0.4-0.6%, e.g., 0.49% w / w citric acid (e.g., citric acid monohydrate);

[0334] 1-3%, e.g., 1.5-2.5%, e.g., 2-3%, e.g., 2.5% w / w sucrose;

[0335] 0.1-1%, e.g., 0.2-0.9%, e.g., 0.4-0.7%, e.g., 0.6% w / w zinc gluconate; and

[0336] 2.5-12.5%, e.g., 5-10%, e.g., 6-9%, e.g.. 7.6% w / w HPMC.

[0337] 266. A pharmaceutical composition comprising:

[0338] 1-4%, e.g., 1.5-3.5%, e.g.. 2-3%, e.g., 2.5% w / w quinine;

[0339] 1-4%. e.g., 1.5-3.5%, e.g.. 2-3%, e.g.. 2.5% w / w EGCG;

[0340] 15-35%, e.g., 20-30%, e.g., 22.5-27.5%, e.g., 24.81% w / w PEG 400;

[0341] 25-55%, e.g., 30-50%, e.g., 35-45%, e.g., 40% w / w PEG 3350;

[0342] 12.5-27.5%, e.g., 15-25%, e.g., 17.5-22.5%, e.g., 20% w / w PEG 400 stearate;

[0343] 0.1-1%, e.g., 0.2-0.8%, e.g., 0.3-0.7%, e.g., 0.4-0.6%, e.g., 0.49% w / w citric acid (e.g., citric acid monohydrate);

[0344] 1-3%, e.g., 1.5-2.5%, e.g., 2-3%, e.g., 2.5% w / w sucrose;

[0345] 0.1-1%, e.g., 0.2-0.9%, e.g., 0.4-0.7%, e.g., 0.6% w / w zinc gluconate; and

[0346] 2.5-12.5%, e.g., 5-10%, e.g., 6-9%, e.g.. 6.6% w / w HPMC.

[0347] 267. A pharmaceutical composition comprising:

[0348] 1-9%. e.g., 3-7%, e.g., 4-6%, e.g.. 5% w / w quinine;

[0349] 1-9%. e.g., 3-7%, e.g., 4-6%, e.g.. 5% w / w EGCG;

[0350] 10-30%, e.g., 15-25%, e.g., 17.5-22.5%, e.g., 19.21% w / w PEG 400;

[0351] 25-55%, e.g., 30-50%, e.g., 35-45%, e.g., 40% w / w PEG 3350;

[0352] 12.5-27.5%, e.g., 15-25%, e.g., 17.5-22.5%, e.g., 20% w / w PEG 400 stearate; 0.1-1%, e.g., 0.2-0.8%, e.g., 0.3-0.7%, e.g., 0.4-0.6%, e.g., 0.49% w / w citric acid (e.g., citric acid monohydrate);

[0353] 1-3%, e.g., 1.5-2.5%, e.g., 2-3%, e.g., 2.5% w / w sucrose;

[0354] 0.6-1.8%, e.g., 0.8-1.6%, e.g., 1-1.4%, e.g., 1.2% w / w zinc gluconate; and

[0355] 2.5-12.5%, e.g., 5-10%, e.g., 6-9%, e.g., 6.6% w / w HPMC.

[0356] 268. A pharmaceutical composition comprising;

[0357] 0.1-2%, e.g., 0.5-1.5%, e.g., 0.75-1.25%, e.g., 1% w / w quinine;

[0358] 0.1-2%, e.g., 0.5-1.5%, e.g.. 0.75-1.25%, e.g., 1% w / w EGCG;

[0359] 20-40%, e.g., 15-35%, e.g., 27.5-32.5%, e.g., 28.29% w / w PEG 400;

[0360] 25-55%, e.g., 30-50%, e.g., 35-45%, e.g., 40% w / w PEG 3350;

[0361] 12.5-27.5%, e.g., 15-25%, e.g., 17.5-22.5%, e.g., 20% w / w PEG 400 stearate;

[0362] 0.1-1%, e.g., 0.2-0.8%, e.g., 0.3-0.7%, e.g., 0.4-0.6%, e.g., 0.49% w / w citric acid (e.g., citric acid monohydrate);

[0363] 1-3%, e.g., 1.5-2.5%, e.g., 2-3%, e.g., 2.5% w / w sucrose;

[0364] 0.06-0.18%, e.g., 0.08-0.16%, e.g., 0.1-0.14%, e.g., 0.12% w / w zinc gluconate; and

[0365] 2.5-12.5%, e.g., 5-10%, e.g., 6-9%, e.g., 6.6% w / w HPMC.

[0366] 269. A pharmaceutical composition comprising;

[0367] 1-10%, e.g., 1.5-8.5%, e.g., 2-6%, e.g., 2.5-5%, e.g., 2.5% or 5% w / w quinine or a salt thereof (e.g., quinine sulfate);

[0368] 1-10%, e.g.. 1.5-8.5%, e.g., 2-6%. e.g., 2.5-5%, e.g., 2.5% or 5% w / w EGCG;

[0369] 15-40%, e.g., 20-35%, e.g., 22.5-27.5%, e.g., 25.16% w / w PEG 400;

[0370] 25-55%, e.g., 30-50%, e.g., 35-45%, e.g., 40% w / w PEG 3350;

[0371] 12.5-27.5%, e.g., 15-25%, e.g., 17.5-22.5%, e.g., 20% w / w PEG 400 stearate (e.g., PEG400 monostearate;

[0372] 0.1-1%, e.g., 0.2-0.8%, e.g., 0.3-0.7%, e.g., 0.4-0.6%, e.g., 0.5% w / w citric acid (e.g., citric acid monohydrate); 1-3%, e.g., 1.5-2.5%, e.g., 2-3%, e.g., 2.5% w / w sucrose:

[0373] 0.05-0.5%, e.g., 0.1 -0.4%, e.g., 0.15-0.3%, e.g., 0.24% w / w zinc acetate; and

[0374] 2.5-12.5%, e.g., 5-10%, e.g., 6-9%, e.g., 6.6% w / w HPMC (e.g., HPMC K100M).

[0375] 270. The pharmaceutical composition of any of embodiments 132-269, which is formulated as a suppository.

[0376] 271. Tire pharmaceutical composition of any of embodiments 132-270, which further comprises epigallocatechin gallate (EGCG).

[0377] 272. The pharmaceutical composition of any of embodiments 132-271, which further comprises quinine or a salt thereof.

[0378] 273. The pharmaceutical composition of any of embodiments 132-272, which further comprises zinc or a salt thereof, e.g., zinc acetate, zinc gluconate or zinc citrate.

[0379] 274. The pharmaceutical composition of any of embodiments 132-273, which further comprises a pharmaceutical composition of any of embodiments 1-131.

[0380] 275. Tire pharmaceutical composition of any of the preceding embodiments, which is formulated for topical administration.

[0381] 276. The pharmaceutical composition of any of the preceding embodiments, which is a solid at 37 C.

[0382] 277. The pharmaceutical composition of any of the preceding embodiments, which does not melt at 37 C.

[0383] 278. The pharmaceutical composition of any of the preceding embodiments, which is a solid at 38, 39, 40, 41, 42, 43, 44, or 45 C.

[0384] 279. Tire pharmaceutical composition of any of the preceding embodiments, which does not melt at 38, 39, 40, 41, 42, 43, 44, or 45 C.

[0385] 280. The pharmaceutical composition of any of the preceding embodiments, which has a mean residence time in the female reproductive tract of 6-9 hours.

[0386] 281. The pharmaceutical composition of any of the preceding embodiments, which has a mean residence time in the female reproductive tract of less than 9, 12, or 24 hours, or less than 2, 3, 4, 5, 6, 7, 14, 21, or 28 days. 282. The pharmaceutical composition of any of the preceding embodiments, wherein the inactive ingredients fully erode in 6 - 9 hours in the female reproductive tract.

[0387] 283. The pharmaceutical composition of any of the preceding embodiments, wherein tire inactive ingredients fully erode in less than 9, 12, or 24 hours, or less than 2, 3, 4, 5, 6, 7, 14, 21, or 28 days in the female reproductive tract.

[0388] 284. The pharmaceutical composition of any of the preceding embodiments, which is a solid at room temperature.

[0389] 285. The pharmaceutical composition of any of the preceding embodiments, which is a solid at room temperature and converts to a gel in the female reproductive tract.

[0390] 286. The pharmaceutical composition of any of the preceding embodiments, which is mucoadhesive in the female reproductive tract.

[0391] 287. A method of treating a cancer, prc-canccr, or infection in a subject in need thereof, comprising administering to the subject an effective amount of a pharmaceutical composition of any of the preceding embodiments.

[0392] 288. Use of a pharmaceutical composition of any of the preceding embodiments in the manufacture of a medicament for treating a cancer or infection in a subject in need thereof.

[0393] 289. A pharmaceutical composition of any of the preceding embodiments for use in treating a cancer or infection in a subject in need thereof.

[0394] 290. A method of preventing a cancer or infection in a subject in need thereof, comprising administering to the subject an effective amount of a pharmaceutical composition of any of the preceding embodiments.

[0395] 291. Use of a pharmaceutical composition of any of the preceding embodiments in the manufacture of a medicament for preventing a cancer or infection in a subject in need thereof.

[0396] 292. A pharmaceutical composition of any of the preceding embodiments for use in preventing a cancer or infection in a subject in need thereof.

[0397] 293. Tire method, use, or composition for use of any of embodiments 287-292, wherein the cancer or infection is mucosal epithelial cancer (e.g., vulvar, vaginal, anal, or oropharyngeal malignancy), an epithelial neoplasia (e.g., cervical dysplasia, genital mucosal neoplasia, oropharyngeal dysplasia, oropharyngeal neoplasia, oral mucosal neoplasia, or anal mucosal neoplasia), cutaneous precancerous and cancerous lesions (e.g., actinic keratosis), a squamous cell cancer, warts (e.g., genital warts), cervical cancer, HPV infection, or HPV induced warts.

[0398] 294. The method, use, or composition for use of any of embodiments 287-292, wherein the pre-cancer is epithelial pre-cancer (e.g., cervical, vulvar, vaginal, anal, urethral, cutaneous, or oropharyngeal dysplasia).

[0399] 295. The method, use, or composition for use of any of embodiments 287-292, wherein the cancer is a mucosal epithelial cancer (e.g., vulvar, vaginal, anal, or oropharyngeal malignancy), cutaneous cancerous lesions, a squamous cell cancer, an adenocarcinoma, or cervical cancer.

[0400] 296. The method, use, or composition for use of any of embodiments 287-295, wherein the cancer or infection is accessible for topical treatment.

[0401] 297. Tire method of any of embodiments 287-295, wherein tire pharmaceutical composition is administered (e.g.. as a first treatment interval) once per day, e.g., for a period of 21 days.

[0402] 298. The method of embodiment 297, wherein the pharmaceutical composition is subsequently administered as a second treatment interval, wherein optionally the second treatment interval comprises administration once per day. e.g., for a period of 21 days.

[0403] 299. The method of any of embodiments 287-298, wherein the pharmaceutical composition is administered (e.g., as a first treatment interval) 1-2 times per week, e.g., for 16 weeks excluding during menses.

[0404] 300. Tire use or composition for use of any of embodiments 287-299, wherein the pharmaceutical composition is fonnulated for administration (e.g., as a first treatment interval) once per day, e.g., for a period of 21 days.

[0405] 301. The use or composition for use of embodiment 300. wherein the pharmaceutical composition is formulated for subsequent administration as a second treatment interval, wherein optionally the second treatment interval comprises administration once per day, e.g., for a period of 21 days.

[0406] 302. The use or composition of embodiment 301, wherein the treatment interval is repeated, e.g., every 6 or 12 months.

[0407] 303. The use or composition for use of any of embodiments 287-302, wherein the pharmacal composition is fonnulated for administration (e.g., as a first treatment interval) 1-2 times per week, e.g.. for 16 weeks excluding during menses. 304. The method, use, or composition for use of any of embodiments 287-303, wherein the subject has previously received surgical ablation (e.g., loop electrosurgical excision procedure (LEEP), resection, conization), cryoablation, or thermal ablation.

[0408] 305. The method, use, or composition of embodiment 304, wherein the subject had, or was identified as having, cervical intraepithelial neoplasia (CIN) or localized cervical cancer.

[0409] 306. The method, use, or composition for use of any of embodiments 287-305, wherein the pharmaceutical composition is administered as a first line therapy.

[0410] 307. The method, use, or composition for use of any of embodiments 287-306, wherein the cervical dysplasia is graded as atypical squamous cells of undetermined significance (ASCUS), atypical squamous cells - cannot exclude high grade squamous intraepithelial lesion (ASC-H), low-grade squamous intraepithelial lesion (LSIL), high-grade squamous intraepithelial lesion (HSIL), cervical intraepithelial neoplasia (CIN) 1. CIN2, or CIN3, or cervical glandular intraepithelial neoplasia (CGIN).

[0411] 308. The method, use, or composition for use of any of embodiments 287-307. wherein the HPV is HPV type 16, 18, 45, or 52.

[0412] 309. The method, use, or composition for use of any of embodiments 287-307. wherein the HPV is HPV type 6, 11, 31, 33, 34, 35, 39, 42, 43, 44, 51, 56, 58, 59, 66, 68, or 70.

[0413] 310. The method, use, or composition for use of any of embodiments 287-309, wherein the subject experiences a selective reduction of viability of dysplastic or neoplastic cells versus surrounding normal cells.

[0414] 311. The method, use, or composition for use of any of embodiments 287-310, which has an EC50 for cervical non-dysplastic cells that higher than the EC50 for cervical dysplastic cells.

[0415] 312. A method of making or manufacturing a pharmaceutical composition of any of the preceding embodiments, the method comprising combining the components of the pharmaceutical composition of any of the preceding embodiments, thereby making or manufacturing the pharmaceutical composition.

[0416] 313. A method of making or manufacturing a pharmaceutical composition, the method comprising combining two or more of (e.g., all of): epigallocatechin gallate (EGCG); quinine or a salt thereof; and zinc or a salt thereof (e.g.. zinc acetate, zinc gluconate, or zinc citrate). 314. The method of embodiment 313, wherein the method further comprises combining one or more excipients with the EGCG, quinine or salt thereof, and / or zinc or salt thereof.

[0417] 315. A method of making or manufacturing a pharmaceutical composition, the method comprising combining two or more of (e.g., all of): one or more PEGs; a hard fat comprising:

[0418] (a) i) a mono-glyceride ester of a fatty acid; ii) a di-glyceride ester of a fatty acid; and iii) a triglyceride ester of a fatty acid; and / or

[0419] (b) one or more PEG stearates, e.g.. PEG monostearates comprising a PEG having a molecular weight in the range of 200-2000, e.g., PEG monostearates comprising a PEG having a molecular weight in the range of 300-1500; hydroxypropyl methylcellulose (HPMC); and citric acid or a salt thereof (e.g., citric acid monohydratc).

[0420] 316. The method of embodiment 315, wherein the method further comprises combining one or more active ingredients with the one or more PEGs. hard fat, HPMC, or citric acid or a salt thereof (e.g., citric acid monohydrate).

[0421] 317. A method of making or manufacturing a pharmaceutical composition, the method comprising combining: one or more PEGs; a hard fat comprising: i) a mono-glyceride ester of a fatty acid; ii) a di -glyceride ester of a fatty acid; and iii) a triglyceride ester of a fatty acid; epigallocatechin gallate (EGCG); quinine or a salt thereof (e.g., quinine sulfate dihydrate); and zinc or a salt thereof (e.g.. zinc acetate, zinc gluconate or zinc citrate).

[0422] 318. A method of making or manufacturing a pharmaceutical composition, the method comprising combining: one or more PEGs: a hard fat comprising: one or more PEG stearates, e.g., PEG monostearates comprising a PEG having a molecular weight in the range of 200-2000, e.g., PEG monostearates comprising a PEG having a molecular weight in the range of 300-1500; epigallocatechin gallate (EGCG); quinine or a salt thereof (e.g., quinine sulfate dihydrate); and zinc or a salt thereof (e.g., zinc acetate, zinc gluconate or zinc citrate).

[0423] 319. Tire method of embodiment 318, wherein the combining is performed simultaneously or sequentially.

[0424] 320. The method of embodiment 319, wherein the combining comprises heating, e.g., heating to about 70 C.

[0425] 321. The method of embodiment 320, wherein the heating is performed in a microwave or water bath.

[0426] 322. The method of any of embodiments 312-321, which further comprises swirling or stirring the pharmaceutical composition.

[0427] 323. Tire method of any of embodiments 312-322, which further comprises adding sucrose or a salt thereof, e.g.. at the heating step.

[0428] 324. The method of any of embodiments 312-323, which further comprises adding citric acid or a salt thereof (e.g., citric acid monohydrate).

[0429] 325. The method of any of embodiments 312-324, which further comprises adding a colorant.

[0430] 326. The method of any of embodiments 312-325, which further comprises adding HPMC, e.g., after the heating step.

[0431] 327. Tire method of embodiment 326, which further comprises stirring or shaking the pharmaceutical composition after addition of the EIPMC.

[0432] 328. The method of any of embodiments 312-327, which further comprises placing the pharmaceutical composition in one or more molds, wherein optionally each mold has a volume of 0.5-4, 1-3, or 1.5-2.5 ml, e.g., 2 ml.

[0433] 329. The method of any of embodiments 312-328, which further comprises allowing the pharmaceutical composition to cool. 330. The pharmaceutical composition, composition, method, or use of any of the preceding embodiments, wherein the zinc salt is zinc acetate.

[0434] 331. The pharmaceutical composition, composition, method, or use of any of the preceding embodiments, wherein the EGCG used to make the pharmaceutical composition is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% pure.

[0435] 332. Tire pharmaceutical composition, composition, method, or use of any of the preceding embodiments, which is substantially free of or does not comprise choline chloride.

[0436] 333. The pharmaceutical composition, composition, method, or use of any of the preceding embodiments, which contains less than 40%, 35%, 30%, 25%, 20%, 15%, or 10% EGCG.

[0437] 334. The pharmaceutical composition, composition, method, or use of any of the preceding embodiments, which comprises less than 200mg, 150 mg, 100 mg, or 75 mg EGCG per unit dose.

[0438] 335. The pharmaceutical composition, composition, method, or use of any of the preceding embodiments, which is not formulated for oral administration.

[0439] 336. Tire pharmaceutical composition, composition, method, or use of any of the preceding embodiments, which is not formulated as a liquid in a capsule.

[0440] DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

[0441] Definitions

[0442] Those skilled in the art will recognize that, in general, terms used herein, and especially in the appended claims (e.g., bodies of the appended claims) are generally intended as "open" terms (e.g., the term "including" should be interpreted as "including but not limited to," the term "having" should be interpreted as "having at least," the term "includes" should be interpreted as "includes but is not limited to." etc.). It will be further understood by those within the art that if a specific number of an introduced claim recitation is intended, such an intent will be explicitly recited in the claim, and in the absence of such recitation no such intent is present. For example, as an aid to understanding, the following appended claims may contain usage of the introductory phrases "at least one" and "one or more" to introduce claim recitations. However, the use of such phrases should not be construed to imply that the introduction of a claim recitation by the indefinite articles "a" or "an" limits any particular claim containing such introduced claim recitation to claims containing only one such recitation, even when the same claim includes the introductory phrases "one or more" or "at least one" and indefinite articles such as "a" or "an' (e.g., "a" and / or "an" should typically be interpreted to mean "at least one" or "one or more"); the same holds true for the use of definite articles used to introduce claim recitations.

[0443] In addition, even if a specific number of an introduced claim recitation is explicitly recited, those skilled in the art will recognize that such recitation should typically be interpreted to mean at least the recited number (e.g., the bare recitation of "two recitations," without other modifiers, typically means at least two recitations, or two or more recitations). Furthermore, in those instances where a convention analogous to "at least one of A, B, and C, etc." is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., "a system having at least one of A, B, and C" would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). In those instances where a convention analogous to "at least one of A, B, or C, etc." is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., "a system having at least one of A, B, or C" would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). It will be further understood by those within the art that typically a disjunctive word and / or phrase presenting two or more alternative tenns, whether in the description, claims, or drawings, should be understood to contemplate the possibilities of including one of the terms, either of the terms, or both terms unless context dictates otherwise. For example, the phrase "A or B" will be typically understood to include the possibilities of "A" or "B" or "A and B."

[0444] With respect to the appended claims, those skilled in the art will appreciate that recited operations therein may generally be performed in any order. Also, although various operational flow diagrams may be presented in a sequence(s), it should be understood that the various operations may be perfonned in other orders than those which are illustrated, or may be perfonned concurrently. Examples of such alternate orderings may include overlapping, interleaved, intermpted, reordered, incremental, preparatory, supplemental, simultaneous, reverse, or other variant orderings, unless context dictates otherwise. Furthermore, terms like "responsive to," "related to," or other past-tense adjectives are generally not intended to exclude such variants, unless context dictates otherwise.

[0445] It is worthy to note that any reference to "one aspect," "an aspect," "an exemplification," "one exemplification." and the like means that a particular feature, structure, or characteristic described in connection with the aspect is included in at least one aspect. Thus, appearances of the phrases "in one aspect," "in an aspect," "in an exemplification," and "in one exemplification" in various places throughout the specification arc not necessarily all referring to the same aspect. Furthermore, the particular features, structures or characteristics may be combined in any suitable manner in one or more aspects. As used herein, the singular form of "a", "an", and "the" include the plural references unless the context clearly dictates otherwise.

[0446] As used herein, the term "comprising" is not intended to be limiting, but may be a transitional term synonymous with "including," "containing," or "characterized by." The temi "comprising" may thereby be inclusive or open-ended and does not exclude additional, unrecited elements or method steps when used in a claim. For instance, in describing a method, "comprising" indicates that the claim is open- ended and allows for additional steps. In describing a device, "comprising" may mean that a named element(s) may be essential for an embodiment or aspect, but other elements may be added and still form a construct within the scope of a claim. In contrast, the transitional phrase "consisting of excludes any element, step, or ingredient not specified in a claim. This is consistent with the use of the term throughout the specification.

[0447] As used herein, the term “EGCG degradant” refers to a compound that is a breakdown product of EGCG exposed to air at room temperature in the absence of an antioxidant other than EGCG. In some embodiments, the EGCG degradant is formed by oxidation of EGCG. In some embodiments, the EGCG degradant is formed by epimerization. In some embodiments, tire EGCG degradant is theasinensin A or (- )-gallocatechin gallate (GCG).

[0448] As used herein, the term 'hard fat” refers to an ester of fatty acid that is solid at a temperature of 18 C. In some embodiments, the hard fat is an ester of fatty acid with glycerol (e.g., with 1 to 3 fatty acids per glycerol molecule). In some embodiments, the hard fat is an ester of fatty acid with PEG or another hydrophilic molecule with free hydroxyl groups. In some embodiments, the hard fat comprises glycerides. In some embodiments, the hard fat comprises triglycerides. In some embodiments, the hard fat comprises mono-, di-, and triglycerides. In some embodiments, tire fatty acid is saturated. In some embodiments, the hard fat has a melting point between 30C and 40C.

[0449] As used herein, "treatment”, "treating" and cognates thereof refer to the medical management of a subject with the intent to improve, ameliorate, stabilize, prevent or cure a disease, pathological condition, or disorder. This term includes active treatment (treatment directed to improve the disease, pathological condition, or disorder), causal treatment (treatment directed to the cause of the associated disease, pathological condition, or disorder), palliative treatment (treatment designed for the relief of symptoms), preventative treatment (treatment directed to preventing, minimizing or partially or completely inhibiting the development of the associated disease, pathological condition, or disorder); and supportive treatment (treatment employed to supplement another therapy).

[0450] Topical treatment of dysplastic or neoplastic cells should feature selective destruction of pre- malignant cells with relative exclusion of normal cells, and with a benign safety profile. A goal of the present invention is to provide a well-tolerated topical treatment for regression of cervical dysplasia with such selectivity.

[0451] Preferred dysplasia-regressing compounds of the invention comprise EGCG and quinine. For codelivery of both agents to an epithelial surface, a solvent system in which both compounds are adequately soluble and stable, and which further enables creation of a suitable pharmaceutical fonnulation must be identified. Dissolution of the suppository and release of quinine and EGCG from the formulation must occur at a rate and extent that facilitates efficient local treatment without displacement or diffusion to other parts of the vagina. Quinine is very poorly soluble in aqueous media, and EGCG is unstable in water, so that nonaqueous solvents and formulations are advantageous.

[0452] Tire invention comprises topical compositions including intravaginal suppositories that concurrently deliver EGCG or, in another embodiment both quinine and EGCG, into dysplastic cells on the cervix or other mucosal tissues displaying dysplasia, including oropharyngeal, vulvar, vaginal or anal epithelium.

[0453] Without being bound to a specific mechanism of action, an important feature of the invention involves selective reduction of viability of dysplastic or neoplastic cells versus surrounding normal cells, leading to regression of precancerous lesions (aka. Dysplasia or CIN) in mucosal epithelium. It is believed that quinine is taken up into, and retained by. dysplastic and neoplastic cells by cation trapping, due to specific changes in the properties of lysosomes in such precancerous and cancerous cells. Quinine has a net neutral charge at pH ~7, which enables permeation of cell membranes. In lower pH environments, such as the interior of lysosomes and other acidic endosomes, quinine has a cationic charge, which limits its egress and thereby enables accumulation in such acidic vacuoles. A stereotyped feature of dysplastic and neoplastic cells is enhanced lysosomal acidification, leading to a lower intralysosomal pH than in corresponding normal cells, with consequent enhanced accumulation of quinine versus non-neoplastic epithelial cells. Sufficient loading of lysosomes with quinine results in impaired cell viability, affecting neoplastic cells more strongly than untransformed cells including those not infected with HPV.

[0454] EGCG (epigallocatechin gallate) is a polyphenolic compound extracted from green tea. While it has been reported to display activity against cervical dysplasia in vitro and in vivo, EGCG is unstable in aqueous media, compromising its potential activity against epithelial mucosal dysplasia and neoplasia. A serious problem with pharmaceutical use of EGCG. which compromises its therapeutic activities, is that it is unstable and undergoes rapid degradation in aqueous media. EGCG may be more stable in oil, petrolatum, fat or wax-based media or hydrophobic excipients such as fatty acid esters (e.g. isopropyl myristate), which arc common ingredients in suppositories, but such excipients can degrade the integrity of latex or polyisoprene condoms, or inhibit use of barrier methods altogether, presenting a problem for prevention of pregnancy and sexually transmissible infections, including HPV.

[0455] A feature of the invention is the discovery that a combination of topically applied quinine and EGCG yields better impairment of viability of dysplastic cells than does either agent alone.

[0456] A primary goal of the invention is to provide pharmacal compositions comprising EGCG and quinine in excipients that improve stability of EGCG for extended shelflife, and which deliver quinine and EGCG at concentrations and for durations sufficient to induce regression of mucosal epithelial dysplasia toward lower grades or non-detectable levels.

[0457] In some embodiments, the pharmacal composition comprises an antioxidant. In some embodiments, the antioxidant is astaxanthin. In some embodiments, tire antioxidant aids in stabilizing and preventing degradation of EGCG.

[0458] In one embodiment, a pharmaceutical composition of the invention comprises intravaginal suppositories that co-deliver quinine and EGCG to the cervix of a subject.

[0459] Co-dclivcry of both quinine and EGCG in a single pharmaceutically-acceptable formulation, including a suppository, must overcome several problems, including poor solubility of quinine in aqueous media and other common solvents used in topical pharmacal formulations. EGCG has stability problems in aqueous or partially aqueous solvent systems. Further goals for fonnulation of intravaginal suppositories include dissolution with minimal unabsorbed material, as well as minimization of osmotic fluid accumulation in the vagina during treatment. Safety is paramount for a self-administered treatment of CIN, so excipients must all have benign safety profiles.

[0460] A suitable pharmacal composition is an intravaginal suppository comprising 2 to 20% EGCG, e.g., 2.5% or 5% (advantageously 5 to 20%, and more specifically 10 to 15%), and 1 to 5% quinine, e.g., 2.5% or 5%, in a pharmaceutically-acceptable excipient base. It is understood that when a composition is described in ranges of percentages of its components, the sum of the percentages is not greater than 100%. Polyethylene glycol (PEG) is an advantageous base, PEG is available in different molecular weights with different melting points and mechanical properties such that defined mixtures of PEGs can be devised to optimize melting point, softness, dissolution rate, and drug release rate and duration. Advantageous PEG molecular weights include but are not limited to 400, 1500 and 4000. It is understood that PEG with a particular molecular weight (e.g., PEG400, PEG1500, PEG3350, or PEG4000) comprises individual PEG molecules with an average molecular weight of that value (e.g.. 400 g / mol, 1500 g / mol, 3350 g / mol, or 4000 g / mol). Generally, each individual PEG molecule does not have an exact molecular weight of, e.g., 400 g / mol, 1500 g / mol, 3350 g / mol, or 4000 g / mol. Typically, there is variation in the molecular weight of each individual PEG molecule. Mixtures of these variants of PEG have appropriate functional properties for formulation of intravaginal suppositories of the invention. An appropriate dose per unit (single suppository for cervical dysplasia, for example), is a minimum of 175 mg of EGCG, more preferably 200 to 500 mg per dose, and more specifically 200 to 400 mg EGCG per dose in a suppository.

[0461] When used in combination with EGCG, a suitable dose of quinine per vaginal suppository for treatment of cervical dysplasia is 40 to 200 milligrams, more specifically 60 to 120 milligrams.

[0462] Mucoadhesive properties are advantageous in a composition for topical treatment of the cervix, to prolong delivery of the active agents directly to the dysplastic lesions in need of treatment. Suitable mucoadhesive excipients which can be melted into a suppository composition include 1) carbopols; 2) cellulose ethers including but not limited to hydroxyethylcellulose, hydroxypropylmethylcellulose, and methylcellulose; 3) vegetable gums including but not limited to gum Arabic and xanthan gum, and 4) polyvinyl alcohol. Single mucoadhesive agents can be used, and also mixtures of two or more mucoadhesive excipients. Suitable concentrations of individual mucoadhesive agents in an intravaginal suppository are in the range of 0.2% to 2%, determined by verification of mucoadhesive properties.

[0463] In one embodiment of the invention, the topical composition further comprises a penetration enhancer to improve entry of the active agents into dysplastic cells. Suitable penetration enhancers include but are not limited to Tween-80, isopropyl myristate, a one, Myrj 52 and transcutol.

[0464] Additional excipients can be used in the suppositories. Lactic acid suitable for buffering the pH to approximately 4 to 5. the natural pH range for a healthy vagina.

[0465] In one embodiment, the present invention is pharmaceutical compositions comprising green tea phytosomes containing EGCG and zinc salt in a pharmaceutically acceptable formulation for topical application to mucosal or cutaneous epithelial surfaces, for elimination of dysplastic or neoplastic cells. In some embodiments, the pharmaceutical composition does not comprise Sanguinaria Canadensis . In some embodiments, the Sanguinaria canadensis may be in solution fomr, e.g., in solution in an alcohol. In some embodiments, the Sanguinaria canadensis is an extract of Sanguinaria canadensis. A primary embodiment is an intravaginal suppository or treatment of cervical dysplasia, for regressing or eliminating cervical intraepithelial neoplasia.

[0466] A suitable size for an intravaginal suppository for treatment of cervical dysplasia is 0.5 to 4 grams, advantageous 1 to 3 grams, and more specifically approximately 2 grams. Hie shape should have rounded edges and an approximately ovoid shape, though any shape that is straightforward to insert and which can adhere to the cervix is suitable.

[0467] Other dosage forms that have the general property of providing locally high concentrations of the active agents at dysplastic patches of the cervical epithelial surface are included within the scope of the disclosure. Compressed tablets comprising the active agents of the disclosure with mucoadhesive polymers, advantageously cellulosic thickeners and flow -enhancing agents for tablet pressing, including but not limited to magnesium stearate or zinc stearate. Thermosetting gels that are relatively soft or liquid at ambient temperature but which thicken at body temperature can be produced with poloxamers combined with mucoadhesive agents such as modified cellulose derivatives in aqueous media. Similar compositions can be lyophilized, providing a solid or semisolid dosage for that hydrate, imparting mucoadhesion and controlled drug delivery.

[0468] For treatment of cervical dysplasia, including HPV-induced ASCUS, CIN1. CIN2 and CIN3, LSIL, and HSIL, a suppository comprising EGCG with or without quinine is inserted into the vagina and placed in contact with the cervix, generally at night before the subject goes to sleep, either using a commercially available suppository applicator or manually with the use of a finger. In some embodiments, the HPV strain that induces the ASCUS, CIN1, CIN2, and / or CIN3, USIU, and HSIL, to be treated is a strain that is discovered or named before or after the filing date of the instant application. In some embodiments, EGCG. quinine or a salt thereof (e.g.. quinine sulfate), and zinc or a salt thereof (e.g.. zinc acetate, zinc gluconate, or zinc citrate) are formulated in a single suppository. In some embodiments, two of EGCG, quinine, and zinc or a salt thereof are formulated together in a first suppository, and the third is formulated in a second suppository. In some embodiments, EGCG and quinine are formulated in a single suppository. In some embodiments, EGCG is formulated in a first suppository, and quinine is formulated in a second suppository. In some embodiments, the suppository is self-administered.

[0469] In some embodiments, the suppository comprises the formulation set forth in Table 24.

[0470] Table 24. Exemplary suppository formulation

[0471] In some embodiments, a suppository comprising EGCG and quinine is used for treatment of cervical dysplasia, including HPV-induced ASCUS, CIN1, CIN2 and CIN3, LSIL, and HSIL, or any disease or disorder described herein. In some embodiments, a first suppository' comprising EGCG and a second suppository comprising quinine is used for treatment of cervical dysplasia, including HPV- induced ASCUS, CIN1, CIN2 and CIN3, LSIL, and HSIL, or any disease or disorder described herein.

[0472] In some embodiments, the suppository' further comprises GMS (glycerol monostearate).

[0473] In some embodiments, the system used to classify cervical dysplasia is the Bethesda system. In some embodiments, the cervical dysplasia is identified as ASC-US (atypical squamous cells of undetermined significance). In some embodiments, the cervical dysplasia is identified as ASC-H (atypical squamous cells cannot exclude HSIL). In some embodiments, the cervical dysplasia is identified as LSIL (low-grade squamous intraepithelial lesions).

[0474] In some embodiments, the system used to classify cervical dysplasia is the CIN (cervical intraepithelial neoplasia) system. In some embodiments, the cervical dysplasia is identified as HSIL (high-grade squamous intraepithelial lesions, moderately to severely abnormal squamous cells). In some embodiments, the cervical dysplasia is identified as CIN1 (e.g., mild dysplasia). In some embodiments, the cervical dysplasia is identified as CIN2 (e.g., moderate dysplasia). In some embodiments, the cervical dysplasia is identified as CIN3 (e.g., severe dysplasia).

[0475] In some embodiments, the cervical dysplasia affects both squamous and glandular cells. In some embodiments, the cervical dysplasia is identified as CGIN (cervical glandular intraepithelial neoplasia). In some embodiments, the CGIN is identified as AGC-NOS (atypical glandular cells not otherwise specified). In some embodiments, the CGIN is identified as AGC-N (atypical glandular cells favoring neoplasia). In some embodiments, the CGIN is identified as AIS (adenocarcinoma in situ).

[0476] In some embodiments, the subject has a chronic HPV infection.

[0477] A suitable regimen begins shortly after cessation of a menstrual period, w ith daily administration, generally self-administration for 21 days, optionally followed by a cervical examination for determination of dysplasia grade and extent. If the resulting grade and extent of dysplasia is judged to be low -risk, no Anther immediate treatment may be necessary, though the 21 -day regimen of selfadministration of intravaginal suppositories of the invention can be repeated periodically, for example, once every 6 months or once every year. An additional intended effect of treatment, particularly of higher-grade dysplasia such as CIN2 or CIN3 or HSIL, if full regression of dysplasia is not achieved via use of intravaginal suppositories, is to reduce the severity and area of dysplastic cells to minimize the intensity and breadth of a subsequent ablative treatment required to eliminate dysplastic cells (if such a procedure is deemed necessary), to minimize adverse consequences of such treatment, such as damage to the integrity of the cervix, which is known to impair various aspects of pregnancy. Any of several ablative modalities can in principle be used in this setting, including thermal ablation, LEEP (Loop Elcctrosurgical Excision Procedure), or surgical or laser conization; how ever, an advantageous ablative modality consistent with the critical goal of minimizing damage to the cervix, is photodynamic therapy, preferably with quinine and riboflavin as photosensitizers activated by light at a wavelength of 405 nm. The pretreatment with intravaginal suppositories delivering quinine specifically for approximately 21 days results in improved efficacy of a subsequent photodynamic therapy session, by enabling progressive accumulation of quinine in dysplastic cells, especially within their lysosomes.

[0478] Quinine accumulates with some selectivity in dysplastic cells, and is also fluorescent with illumination under ultraviolet or, preferably short wavelength visible light, such as 405 nm light. Therefore, illumination of the cervix with 405 nm light during examination during colposcopy examination can visualize residual areas of dysplasia due to their accumulation of quinine during treatment.

[0479] Quinine and EGCG selectively reduce viability in dysplastic and neoplastic cells via fundamentally different mechanisms, so that combined use of these agents can enhance both selectivity and anti -dysplastic activity versus either agent alone.

[0480] In some embodiments, the present invention is pharmaceutical compositions comprising EGCG or EGCG combined with quinine in a pharmaceutically acceptable formulation for topical application to mucosal or cutaneous epithelial surfaces, for elimination of dysplastic or neoplastic cells. A primary embodiment is an intravaginal suppository or treatment of cervical dysplasia, for regressing or eliminating cervical intraepithelial neoplasia with a suppository composition as described herein.

[0481] Treatment of EIPV -induced or other neoplastic conditions on areas of the body more accessible than the cervix can be accomplished with compositions as described herein in the form of topical spreadable semisolid compositions such as ointments, creams or gels. These conditions include but are not limited to surface dysplasias of the vulvar, anal, inguinal, oropharyngeal regions, actinic keratoses and other precancerous skin lesions, and warts, including HPV-induced external genital warts. In some embodiments, the dosage form is a topical ointment, cream, or gel and the indication is a surface dysplasia of the vulvar, anal, inguinal, oropharyngeal regions, precancerous skin lesions (e.g.. actinic keratosis), and warts, (e.g., HPV-induced external genital warts). Semisolid pharmaceutical compositions for treatment of these accessible lesions do not require the formation of an intermediate hydrogel mass (achieved in suppository embodiments by, for example, inclusion of HPMC powder activated by ambient moisture after insertion of the suppository), but may advantageously comprise stabilizing excipients to prevent premature oxidation, discoloration and loss of EGCG. Another important feature for maximizing EGCG stability is minimization of water in the compositions.

[0482] Concentrations of active agents, including EGCG and quinine, in topical semisolid compositions for accessible lesions are similar to those used in exemplified suppository compositions, and concentration ranges of stabilizing excipients (e.g., citric acid or other organic acids, zinc acetate or other pharmacally -acceptable zinc salts, and sucrose) specified for suppository formulations are also appropriate for topical semisolid compositions. The physical bulk of suitable semisolid compositions comprises several alternatives including but not limited to those below.

[0483] Ointments based on PEG are suitable vehicles for treatment of accessible lesions. Typically, a high molecular weight PEG such as PEG 4000 is mixed with a low molecular weight PEG such as PEG 400 while heating to 65 degrees C, in proportions that result in a spreadable ointment. 70% PEG 400 and 30% PEG 4000 are suitable proportions. Blends of PEG variants with MW from 200 to 6000 can be used to produce suitable ointment bases, to which are also added EGCG with or without quinine, plus EGCG- stabilizing excipients of the as described herein. Surfactant agents, including but not limited to PEG- monostearates are optionally used to adjust consistency and absorption.

[0484] Other standard ointment bases including petrolatum known in the art but containing EGCG- stabilizing excipients of the as described herein within concentration ranges established for the suppository compositions are also suitable vehicles for the active agents.

[0485] In some embodiments, a composition (e.g., pharmaceutical composition) as described herein is a nonaqueous topical gel. These include but are not limited to glycerol, propylene glycol, or a mixture of both, in which EGCG is dissolved, with or without a quinine salt. Tire mixture is thickened to a gel consistency with a cellulosic polymer, preferably 1 to 5% ethylcellulose by mixing and heating to approximately 70 degrees C.

[0486] EXAMPLES

[0487] Example 1

[0488] For preparation of a suppository composition for delivery of EGCG to an epithelial surface, 28 grams of PEG with a molecular w eight of 1500 (PEG 1500) and 12 grams of PEG4000 were mixed and melted together in a microwave oven during heating for 30 seconds, gauging the time to be the minimum required for melting, in the case of different ovens. 10 grams of EGCG, and 500 mg of Carbopol 934 were dissolved in the melted PEG mixture, and poured into suppository molds with a volume of approximately 2 ml per suppository. The suppositories were allowed to cool in a freezer while still in the molds, and then removed from the molds. The composition can be scaled to larger batch sizes, and mixed by either microw ave irradiation or by heating the vessel in which they are mixed.

[0489] Example 2

[0490] For preparation of a suppository composition for co-delivery of quinine and EGCG to an epithelial surface, 28 grams of PEG with a molecular weight of 1500 (PEG1500) and 12 grams of PEG4000 were mixed and melted together in a microwave oven during heating for 30 seconds. 2 grams of quinine hydrochloride, 10 grams of EGCG, and 500 mg of Carbopol 934 were dissolved in the melted PEG mixture, and poured into suppository molds with a volume of approximately 2 ml per suppository . Tire suppositories were allowed to cool in a freezer while still in the molds, and then removed from the molds. The composition can be scaled to larger batch sizes, and mixed by either microwave irradiation or by heating the vessel in which they are mixed.

[0491] Example 3

[0492] For treatment of cervical dysplasia, a suppository of the invention is inserted into the vagina in contact with the cervix of a subject displaying ASCUS or CINE CIN2 or CIN3 abnormalities . A preferred time of day for treatment is shortly before the subject goes to sleep for the night. Treatment is repeated daily for 21 days, beginning shortly after cessation of a menstrual period. A tampon is optionally used to retain the suppository in close apposition to the cervix. The result is regression of dysplastic lesions, both in grade and in size. If a treated lesion is judged to be low-risk for progression toward cervical cancer, no further treatment may be needed, or the 21 -day regimen is repeated at intervals of 6 or 12 months. In the event of insufficient regression, residual dysplasia (with smaller area and lower dysplasia grade than at the beginning of suppository treatment) is also optionally treated with another course of 21 -day suppository treatment, or with an ablative therapy, advantageously photodynamic therapy with a mixture of quinine and riboflavin as photosensitizers, activated by light at a wavelength of 405 nm.

[0493] Example 4

[0494] For preparation of a suppository composition for deli ven of EGCG to an epithelial surface, the intravaginal suppository comprises 100 - 250 mg green tea phytosomc containing a minimum of 13% EGCG, 2 g unrefined coconut oil, 0.5 g aloe barbadensis leaf gel, and 0.25 pl melaleuca altemifolia extract. Coconut oil was melted in saucepan and ingredients added and thoroughly mixed before placing in a designated suppository mold and freezing overnight. Intravaginal suppositories were inserted 1-2 times per week for 16 weeks, excluding during menses, after CIN status was determined to be CIN2 / 3 in a human subject, first by the detection of an abnormal Pap test combined with molecular diagnostic testing which indicated HR-HPV, followed by colposcopy and biopsies which confirmed tire presence of CIN2 / 3. Regression to CIN1 was determined and no ablative therapies or excisional procedures were performed. Example 5

[0495] For preparation of a suppository composition for delivery of EGCG and zinc salt to an epithelial surface, the intravaginal suppository comprises 100 - 250 mg green tea phytosome containing a minimum of 13% EGCG, 1 mg zinc salt. 2 g unrefined coconut oil, 0.5 g aloe barbadensis leaf gel, 12.5 mg activated carbon, and 0.25 pl melaleuca altemifolia extract. Coconut oil was melted in saucepan and ingredients added and thoroughly mixed before placing in a designated suppository mold and freezing overnight. CIN status in a human subject was determined as CIN1 at previous Pap screening with colposcopy and biopsy confirmation two years prior, and after 3 weeks of alternate -evening administration before bedtime of the green tea phytosomc / EGCG and zinc salt suppositories, subsequent Pap, colposcopy, and molecular diagnostic testing for HPV all indicated no dysplasia.

[0496] While the foregoing written description of the invention enables one of ordinary skill to make and use what is considered presently to be the best mode thereof, those of ordinary’ skill will understand and appreciate the existence of variations, combinations, and equivalents of the specific embodiment, method, and examples herein. The invention should therefore not be limited by the above-described embodiment, method, and examples, but by all embodiments and methods within the scope and spirit of the invention.

[0497] Example 6. Dose response of cells to EGCG alone and combined with zinc gluconate in vitro

[0498] This Example demonstrates determination of the IC50 of EGCG alone and in combination with zinc gluconate in vitro in three cell lines.

[0499] IC50 of EGCG in vitro

[0500] Three cell lines were utilized: SiHa (derived from HPV16+ squamous cell carcinoma), CaSki (derived from HPV16+ epithelial carcinoma), and DoTc2-4510 (derived from cervical adenocarcinoma). Cells were grown in a 1: 1 (v:v) mixture of Dulbecco's modified Eagle's medium (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) and Ham's F12 nutrient mix (Nissui Pharmaceutical Co., Ltd., Tokyo. Japan) containing 5% fetal bovine serum (FBS) (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) or no FBS at 37°C in a humidified atmosphere containing 5% CO2. A stock solution of EGCG was made by dissolving EGCG in DMSO. Cells were exposed to EGCG at a concentration of 0, 10, 25, 50, 100, or 250 pg / mL for 48 hours. Cell viability was measured at 0 hours, 24 hours, and 48 hours post treatment. Cells treated with EGCG were re-treated at 48 hours after initial treatment. Hie IC50 of EGCG in was detennined both with (5% FBS) and without (0% FBS) serum in media.

[0501] Cell viability was determined using the CellTiter-Glo® assay (Promega). IC50 is defined as the inflection point on the dose response curve (half of the distance between the min and max of the curve fit). NA denotes that the maximum of the curve is below 50% viability, or the IC50 is otherwise unable to be calculated. Results are shown in Table 1.

[0502] Table 1. IC50 of EGCG in vitro in three cell lines These results show the IC50 of EGCG in three cell lines derived from squamous cell carcinoma, epithelial cell carcinoma, and adenocarcinoma cells.

[0503] IC50 of EGCG combined with zinc gluconate in vitro

[0504] Cells were cultured as described above. In addition to EGCG treatment as described above, cells were treated with 20, 50, 100, 150, 200, or 250 pM zinc gluconate. Results are shown in Tables 2-4.

[0505] Table 2. IC50 of EGCG combined with zinc gluconate in vitro 0 hours after treatment

[0506] Table 3. IC50 of EGCG combined with zinc gluconate in vitro 24 hours after treatment

[0507] * = % viability at lowest concentration of EGCG < 50%

[0508] = unable to determine

[0509] Table 4. IC50 of EGCG combined with zinc gluconate in vitro 48 hours after treatment

[0510] * = % viability at lowest concentration of EGCG < 50%

[0511] These results show the IC50 of EGCG in combination with zinc gluconate in three cell lines derived from squamous cell carcinoma, epithelial cell carcinoma, and adenocarcinoma cells. At 48 hours after treatment, the IC50 of EGCG combined with zinc gluconate is similar to or lower than the IC50 of EGCG alone 48 hours after treatment.

[0512] Example 7. Dose response of cells to quinine alone and EGCG combined with quinine in vitro

[0513] This Example demonstrates determination of the IC50 of EGCG in combination with quinine in vitro in three cell lines.

[0514] IC50 of quinine in vitro

[0515] Three cell lines were utilized: SiHa (derived from HPV16+ squamous cell carcinoma), CaSki (derived from HPV16+ epithelial carcinoma), and DoTc2-4510 (derived from cervical adenocarcinoma). Cells were grown in a 1: 1 (v:v) mixture of Dulbecco's modified Eagle's medium (Sigma- Aldrich; Merck KGaA, Darmstadt, Germany) and Ham's F12 nutrient mix (Nissui Pharmaceutical Co., Ltd., Tokyo, Japan) containing 5% fetal bovine serum (FBS) (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) or no FBS at 37°C in a humidified atmosphere containing 5% CO2. A stock solution of quinine was made by dissolving quinine in DMSO. Cells were exposed to quinine at a concentration of 0, 10, 25, 50, 100, 250, or 500 pg / mL for 48 hours. Cell viability was measured at 0 hours, 24 hours, and 48 hours post treatment. Cells treated with quinine were re-treated at 48 hours after initial treatment. The IC50 of quinine was determined both with (5% FBS) and without (0% FBS) serum in media.

[0516] Cell viability was determined using the CellTiter-Glo® assay (Promega). IC50 is defined as the inflection point on the dose response curve (half of the distance between the min and max of the curve fit). NA denotes that the maximum of the curve is below 50% viability, or the IC50 is otherwise unable to be calculated. Results are shown in Table 5.

[0517] Table 5. IC50 of quinine in vitro in three cell lines

[0518] These results show the IC50 of quinine in three cell lines derived from squamous cell carcinoma, epithelial cell carcinoma, and adenocarcinoma cells.

[0519] IC50 of EGCG combined with quinine in vitro

[0520] Cells were cultured as described above. A stock solution of EGCG was made by dissolving EGCG in DMSO. Cells were treated with 0, 10, 25, 50, 100, or 250 pg / mL EGCG. In addition to EGCG treatment, cells were treated with 25, 63, 126, 252, 630, or 1260 pM quinine. Cells were retreated with EGCG and quinine at 48 hours. The IC50 of EGCG combined with quinine was determined both with (5% FBS) and without (0% FBS) serum in media. Results are shown in Tables 6-8.

[0521] Table 6. IC50 of EGCG combined with quinine in vitro 0 hours after treatment

[0522] Table 7. IC50 of EGCG combined with quinine in vitro 24 hours after treatment

[0523] * = % viability at lowest concentration of EGCG < 50%

[0524] Table 8. IC50 of EGCG combined with quinine in vitro 48 hours after treatment

[0525] * = % viability at lowest concentration of EGCG < 50% Tables 6-8 demonstrate that combining EGCG and quinine lowers the IC50 compared to the IC50 of EGCG or quinine alone. In addition, the combination of EGCG and quinine induced complete loss of viability of all 3 HPV-induced cervical cancer cell lines at certain concentrations, with similar IC50 values in all cell lines. Example 8. Dose response of cells to quinine combined with zinc gluconate in vitro

[0526] This Example demonstrates determination of the IC50 of quinine combined with zinc gluconate in vitro in three cell lines.

[0527] Three cell lines were utilized: SiHa (derived from HPV16+ squamous cell carcinoma), CaSki (derived from HPV16+ epithelial carcinoma), and DoTc2-4510 (derived from cervical adenocarcinoma). Cells were grown in a 1: 1 (v:v) mixture of Dulbecco's modified Eagle's medium (Sigma- Aldrich; Merck KGaA, Darmstadt, Germany) and Ham's F12 nutrient mix (Nissui Pharmaceutical Co., Ltd., Tokyo. Japan) containing 5% fetal bovine serum (FBS) (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) or no FBS at 37°C in a humidified atmosphere containing 5% CO2. A stock solution of quinine was made by dissolving quinine in DMSO. Cells were exposed to quinine at a concentration of 0, 10, 25, 50, 100, 250, or 500 pg / mL and zinc gluconate at a concentration of 0, 20, 50, 100, 150. 200, or 250 pM for 48 hours. Cell viability was measured at 0 hours, 24 hours, and 48 hours post treatment. Cells were retreated at 48 hours after initial treatment. The IC50 was determined both with (5% FBS) and without (0% FBS) serum in media.

[0528] Cell viability was determined using the CellTiter-Glo® assay (Promega). IC50 is defined as the inflection point on the dose response curve (half of the distance between the min and max of the curve fit). NA denotes that the maximum of the curve is below 50% viability, or the IC50 is otherwise unable to be calculated. Results are shown in Tables 9-11.

[0529] Table 9. IC50 of quinine combined with zinc gluconate in vitro 0 hours after treatment

[0530] Table 10. IC50 of quinine combined with zinc gluconate in vitro 24 hours after treatment

[0531] * = % viability at lowest concentration of quinine < 50% Table 11. IC50 of quinine combined with zinc gluconate in vitro 48 hours after treatment

[0532] * = % viability at lowest concentration of quinine < 50%

[0533] Tables 9-11 demonstrate that combining quinine and zinc gluconate generally lowers the IC50 in the three cell lines tested compared to the IC 0 of zinc gluconate or quinine alone.

[0534] Example 9. Dose response of cells to quinine combined with EGCG and zinc gluconate in vitro

[0535] This Example demonstrates determination of the IC50 of quinine combined with EGCG and zinc gluconate vitro in three cell lines.

[0536] Three cell lines were utilized: SiHa (derived from HPV16+ squamous cell carcinoma), CaSki (derived from HPV16+ epithelial carcinoma), and DoTc2-4510 (derived from cervical adenocarcinoma). Cells were grown in a 1: 1 (v:v) mixture of Dulbecco's modified Eagle's medium (Sigma- Aldrich; Merck KGaA, Darmstadt, Germany) and Ham's F12 nutrient mix (Nissui Pharmaceutical Co., Ltd., Tokyo, Japan) containing 5% fetal bovine serum (FBS) (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) or no FBS at 37°C in a humidified atmosphere containing 5% CO2. A stock solution of EGCG was made by dissolving EGCG in DMSO. A stock solution of quinine was made by dissolving quinine in DMSO. Cells were treated with quinine at 0, 10. 25. 50. 10, 250, or 500 pg / mL in combination with:

[0537] Combination 1 : 25 pM EGCG, 25 pM zinc

[0538] Combination 2: 50 pM EGCG, 50 pM zinc

[0539] Combination 3: 75 pM EGCG, 75 pM zinc

[0540] Combination 4: 100 pM EGCG, 100 pM zinc

[0541] Combination 5: 125 pM EGCG, 125 pM zinc

[0542] Combination 6: 50 pM EGCG, 100 pM zinc.

[0543] Cell viability was measured at 0 hours, 24 hours, and 48 hours post treatment. Cells were retreated with quinine and EGCG 48 hours after initial treatment. The IC50 was determined without (0% FBS) serum in media. Cell viability was determined using the CellTiter-Glo® assay (Promega). Cell viability in treated and untreated control wells were measured, and % viability was calculated in treated wells vs untreated control wells. IC50 is defined as the inflection point on the dose response curve (half of the distance between the min and max of the curve fit). NA denotes that the maximum of the curve is below 50% viability, or the IC50 is otherw ise unable to be calculated. Results are shown in Tables 12-17.

[0544] Table 12. IC50 of quinine combined with EGCG and zinc gluconate in vitro 0 hours after treatment

[0545] Table 13. Cell % viability at 0 hours after treatment Table 14. IC50 of quinine combined with EGCG and zinc gluconate in vitro 24 hours after treatment

[0546] * = % viability at lowest concentration of quinine < 50%

[0547] Table 15. Cell % viability at 24 hours after treatment

[0548] Table 16. IC50 of quinine combined with EGCG and zinc gluconate in vitro 48 hours after treatment

[0549] * = % viability at lowest concentration of quinine < 50%

[0550] Table 17. Cell % viability at 48 hours after treatment Tables 12, 14, and 16 show that the combination of EGCG, quinine, and zinc gluconate generally results in lower IC50 values at 24 and 48 hours than EGCG, quinine, or zinc gluconate alone. Tables 13, 15, and 17 show that cells treated with the combination of EGCG, quinine, and zinc gluconate exhibit low viability at 48 hours after treatment. Example 10: Exemplary suppository formulations comprising combinations of PEG, hard fat, and excipients for EGCG stabilization display thermal stability, mucoadhesion, and chemical stability

[0551] This example describes the production of exemplary’ suppository formulations and measurement of their properties. Polyethylene glycol (PEG), in a 1:2:2 mixture of three variants with different molecular weights (average MW 400, 1500 and 4000 respectively) formed a waxy solid material that can dissolve by locally-produced fluids, but did not melt at body temperature. Addition of a hard fat excipient promoted incorporation of the lipophilic active agent and. although this excipient melts at body temperature, when combined with PEG and a surfactant (in this case HPMC, which did not gel in PEG or Ovucire, but imparted mucoadhesive properties when it hydrated after application), it yielded a relatively- heat-stable material that underwent gradual dissolution and gelation (due to HPMC hydration) when warmed to body temperature (37 degrees C) in the presence of moisture, simulating the cervical environment.

[0552] A batch of about 38 grams of suppository material was made using:

[0553] • PEG 400, 5 g

[0554] • PEG 1500, 10 g

[0555] • PEG 4000, 10 g

[0556] • Gattefosse Ovucire WL 3264, 8 g

[0557] • Citric acid 200 mg

[0558] • Sucrose 1 g

[0559] • Zn Gluconate. 0.25 g

[0560] • EGCG, 1 g

[0561] • Quinine HCL, 1 g

[0562] • HPMC (K100M), 3 g

[0563] Polyethylene glycol (PEG)

[0564] Polyethylene glycol (PEG) is polymer that is available in preparations with different average molecular weights (MW). Tirus, PEG 400 is a liquid with an average molecular weight of 400 daltons, and PEG 1500 and PEG 4000 are solids with MW of 1500 and 4000 respectively. Different varieties of PEG can be blended to yield materials with unique properties not shared by PEG of a specific single average MW. Through experimentation, a combination of PEG 400, PEG 1500 and PEG 4000 in ratios of 1:2:2 was found to have suitable properties for vaginal suppositories of the disclosure, including thermal stability at elevated storage temperatures up to approximately 40 degrees C.

[0565] Hard Fat

[0566] Suitable base ingredients for suppositories comprise compounds broadly described as hard fat, which can include glycerides (fatty acids esterified to glycerol, with 1 to 3 fatty acids per glycerol molecule) or fatty acid esters of PEG or other hydrophilic molecules with free hydroxyl groups. An exemplary hard fat for suppositories is known as "Ovucire”; Ovucire WL 3264 is a specific blend of waxy hard fat compounds with mild surfactant properties. Ovucire and other hard fats designed for conventional suppositories typically melt at about 30 to 33 degrees C, below body temperature (and also below ambient temperatures that may occur during shipping and storage, which can complicate distribution and deployment of hard fat suppositories).

[0567] Excipients that promote stabilization of EGCG

[0568] EGCG in pharmaceutical formulations can oxidize and undergo color changes (turning brown) and loss of activity during storage, limiting its utility as an active pharmaceutical ingredient. Patient acceptance and compliance can be compromised by use of a brown-colored formulation that can stain undergarments. This disclosure provides, among other things, excipients that inhibit EGCG oxidation and color change. Citric acid, sucrose and a source of divalent zinc ions, in this case, zinc gluconate, were identified as suitable stabilizing excipients, with expected benign safety profiles. All three agents combined together gave a superior stabilizing activity compared to any of the agents alone, during storage at elevated temperatures.

[0569] Excipients that promote mucoadhesion

[0570] The suppositories described herein display mucoadhesion. in which suppositories applied to the cervix adhere to the cervical epithelium and gradually erode while releasing active agent locally. Conventional suppositories are typically formulated for rapid melting or dissolution, delivering active agents throughout the vaginal cavity, as they are generally intended for either distribution over a wide area, for example for treatment of a yeast infection or for systemic absorption via a large area of mucosal tissue. In contrast, mucoadhesion is important for localized treatment of cervical dysplasia and other diseases or disorders described herein to maximize drug activity in the lesions or abnormal cells, while also maximizing safety by limiting spread to unaffected areas of the vaginal cavity. Through extensive experimentation, variants of the cellulosic polymer hydroxypropyl methyl cellulose (HPMC) were found to impart these desired properties to the suppositories. An important feature of HPMC for manufacturing suppositories of this disclosure is that it did not thicken substantially when incorporated into melted PEG or Ovucire (or mixtures of both excipients), and thus promotes straightforw ard production at both small and large scales. An exemplary variant of HPMC designated K100M (with an average molecular weight of approximately 100.000) promoted simple manufacturing and mucoadhesion. In some embodiments, the viscosity of HPMC KI OOM is beneficial for formulation of the suppository for the reasons described herein. Active Ingredients: EGCG and Quinine

[0571] Although certain active agents described herein for treatment of cervical dysplasia, EGCG and quinine, have different physical and chemical properties, this suppository formulation promoted incorporation of both compounds, as well as their release during simulated use.

[0572] Preparation of the suppository

[0573] The polyethylene glycols, Ovucire WL 3264, EGCG and quinine were weighed out. mixed together, and stirred in a single beaker. These ingredients were heated to 70 degrees C, yielding a homogeneous, thick, pourable liquid. The mild surfactant activity of Ovucire was used to promote blending with the hydrophilic PEG mixture without separation into layers.

[0574] After the other ingredients melted. HPMC was added and stirred, yielding a uniform suspension. The mixture was poured into suppository molds (holding approximately 2 grams each) and chilled in a freezer until the suppositories solidified. The suppositories were then removed from the molds and tested for various properties relevant to their intended clinical use.

[0575] Thermal Stability

[0576] Suppositories were found to retain their physical form and appearance when heated to 40 degrees C, which is higher than the melting point of Ovucire or other hard fats used as suppository bases.

[0577] Mucoadhesion

[0578] Suppositories were placed in individual silicone dishes (weigh boats). A volume of water equivalent to the weight of the suppository (2 ml H2O for 2 gram suppository samples) was added and the suppositories were incubated at 37 degrees C for various times. The suppositories were physically stable at 37 C when dry. In water, the suppositories began to erode, as intended. The suspended HPMC particles began to gel in the presence of water, yielding a mass of soft gel exceeding the dimensions of the original suppository material. For assessment of mucoadhesive properties, the dishes were inverted at various times after addition of water. Adhesion of the suppository material was determined by whether or not it fell out of the inverted dishes.

[0579] One hour after addition of water, the submerged portion of the suppository had begun to erode, with a rim of gel formed by hydration of HPMC. At time points of 2 through 24 hours, the suppository material remained in the silicone dishes when inverted, indicating strong adhesive properties.

[0580] Chemical stability’

[0581] Suppositories comprising EGCG, in the absence of a stabilizer, began as an off-white material that gradually turned brown over a period of days to weeks, both at ambient temperature and more rapidly at 37 degrees C. EGCG can undergo autooxidation and irreversible dimerization and then oligomerization resulting in polycyclic phenolic compounds that have a brown color under visible light (these oligomers also account for the conversion of fresh green tea leaves into “black tea” by aging). Visible color changes provide an extremely sensitive index for detecting and quantifying formation of dimers due to autooxidation. Formation of brown oligomers is the primary reaction by which pharmacologically active EGCG is degraded during storage of nonstabilized EGCG compositions. Suppositories containing EGCG in the presence of sucrose and zinc gluconate (but not citric acid) were brown 1 month after formulation and dark brown 3 months after formulation. Suppositories containing EGCG in the presence of sucrose and citric acid (but not zinc gluconate) were beige 1 month after formulation and dark beige 3 months after formulation. Suppositories containing EGCG in the presence of citric acid and zinc gluconate (but not sucrose) were off-white 1 month after formulation and light beige 3 months after formulation.

[0582] Suppositories containing citric acid, sucrose and zinc gluconate did not undergo a color change, even after 3 months at 37 degrees C.

[0583] Table 18. Color change in exemplary suppository formulations

[0584] As Table 18 shows, all three stabilizing excipients contributed to prevention of color changes attributable to EGCG oxidation during storage for 3 months at a temperature of 37 C. Example 11: Exemplary suppository formulations display mucoadhesion and thermal stability

[0585] Five exemplary suppository formulas were prepared to determine the effects of various excipients on thermal stability and formation of a mucoadhesive gel upon contact with aqueous fluid. Compositions are indicated in Table 19. For each formula, all ingredients except for HPMC or HEC were melted together at 70 C and mixed by stirring. HPMC or HEC was added as appropriate and stirred to homogeneity. Suppository material was poured into molds and chilled to harden.

[0586] Table 19. Exemplary suppository formulations

[0587] PEG = Polyethylene glycol Hard Fat = Ovucire WL3264

[0588] GMS = Glycerol monostearate

[0589] PEGoxol-7 Stearate = Stearic acid esterified PEG with an average of 7 monomer units

[0590] HPMC = Hydroxypropyhnethylcellulose

[0591] HEC = Hydroxyethylcellulose Mucoadhesion

[0592] A mucoadhesive study for all five formulae were performed. One suppository of each batch was placed on a plastic weigh boat with 2 mb of water. These weigh boats were then placed in an incubator at 37°C for 24 hours. Pictures of each weigh boat were taken to compare the gelling of the water. Contact with water caused a gel to form in all the suppositories, and all suppositories adhered to their respective weigh boats after inverting the weigh boat at T= 24 hours.

[0593] Thermal Stability

[0594] To test the physical stability of the suppositories in high temperature conditions, suppositories were placed in a 20 mL glass vial and heated in 40°C and 50°C ovens. These temperatures represent extreme storage conditions that could be encountered during transport or storage. All suppositories fonnulated with HPMC maintained their shape and did not melt at 40°C after 24 hours. The suppository formulated with HEC (Formula E) started only slightly melting after 24 hours at 40°C. Therefore. HPMC contributes to thermal stability of the suppository material, in addition to providing mucoadhesion. Additionally, all suppositories showed mild melting after 24 hours at 50°C with the exception of the suppository containing GMS (Formula D) that did not melt after 24 hours at 50°C.

[0595] Example 12: Active ingredient concentration in the gels of exemplary suppository formulations

[0596] Vaginal suppositories of the disclosure adhere to the cervical surface after application and undergo erosion and gel fomiation (due to hydration of HPMC in ambient vaginal fluids), resulting in a small mass of hydrogel. The active agents can then be released and distributed into the HPMC hydrogel. Tests were conducted on several suppository variants to assess mucoadhesive gel formation under simulated use, and to measure concentrations of active agents in the gel.

[0597] In this study, six exemplary suppository formulations (Table 20) were prepared. A method was developed for representative suppository dissolution and migration of the active agents EGCG and quinine from the suppository formulations over a 244rour period in PBS (Phosphate Buffered Saline). A volume of PBS equivalent to the weight of each suppository in grams was used for gel formation. For each study sample, the gelling of the PBS solution in the different formulations was observed at initial, 1 hour, 3 hours, 6 hours, and 24 hours after PBS addition . The gelled solution portion that fonned during the study was decanted after 24 hours and evaluated for EGCG and quinine. Table 20 Exemplary suppository formulations

[0598] PEG = Polyethylene glycol

[0599] Hard Fat = Ovucire WL3264 or alternative PEGoxol-7 Stearate = Stearic acid linked to PEG with an average of 7 monomer units; mixture of mono- and di-stearates

[0600] PEG 400-stearate = Stearic acid linked to PEG with a molecular weight of 400 (monostearate)

[0601] HPMC = Hydroxypropylmethylcellulose Active ingredient concentration

[0602] PBS solution was added into a 1 oz glass jar in a 1 : 1 ratio of suppository weight to PBS solution. Suppositories were then gently placed in the 1 oz glass jar. The suppository naturally placed itself on the side of the jar. The closed 1 oz glass jar with suppository and PBS solution was placed in an incubator set at 37°C. The temperature of the incubator was maintained at 36 - 38°C. Suppositories were observed at initial, 1 hour, 3 hours, 6 hours, and 24 hours after PBS addition. After 24 hours, all liquid / gel surrounding the suppository was decanted into a lOmL amber glass vial. A pipette or spatula was used to aid in the decanting of the liquid / gel. The decanted solution was refrigerated (at 2°C to 8°C) if testing was performed on a different day. Quinine was measured using an HPLC UV detector. EGCG was measured using an LCMS assay.

[0603] All suppository formulations displayed adhesion and gelling properties when exposed to PBS for 24 hours.

[0604] Table 21. Analysis of Decanted Gel Portion of Gelled Solutions from Mucoadhesive Study As noted in Table 21, most formulations released about 30 - 40% of each active ingredient over the 24- hour period of the study at 37 °C. The mixture of PEG 400 and PEG 3500 displayed dissolution and gelation properties similar to those of a tripartite mixture of PEG 400, PEG 1500 and PEG 4000.

[0605] Example 13: Penetration of EGCG and Quinine into Mucosal Epithelium

[0606] Treatment of EIPV -infection and any resulting disease or disorderin cervical epithelium with suppositories of the disclosure can involve the following steps:

[0607] 1. A vaginal suppository is inserted and placed in contact with the cervix

[0608] 2. The suppository begins to erode when in contact with ambient vaginal fluids

[0609] 3. Suppository erosion and mixing with fluids enables HPMC in the composition to hydrate and form a gel with mucoadhesive properties for sustained localized drug delivery into the cervical epithelium

[0610] 4. Active agents migrate from the original suppository material into the gel

[0611] 5. Active agents are then delivered directly from the gel into the cervical epithelium

[0612] In vitro simulations of steps 1-4 are illustrated in Example 12. Step 5 was simulated by using an in vitro penetration test (IVPT), with bovine vaginal mucosa as a surrogate for cervical epithelium and underlying stroma. Gel samples derived from each of the suppository fonnulation variants in Example 12 were tested for relative effectiveness for delivery of actives into or through the mucosal tissue.

[0613] A flux study was conducted to determine the rate and extent of in vitro bovine vaginal tissue permeation of quinine, EGCG (epigallocatechin-3-gallate), and the EGCG metabolite gallic acid (together referred to as the “Actives”) into and through bovine vaginal tissue using a Franz diffusion cell system, in which compounds were applied to the epithelial surface of a piece of mucosal tissue; the lower surface was in contact with a small reservoir of phosphate-buffered saline that received compounds that passed all the way through the tissue sample. Tire tissue sample, after dividing into two layers, was extracted for quantitation of EGCG and quinine. Because one route of EGCG metabolism is cleavage of the esterified gallic acid, yielding epigallocatechin (EGC), measurement of both EGCG and gallic acid in tissue and receiving vessel provides a better estimate of EGCG delivered into mucosal tissue than does measurement EGCG alone. EGCG can undergo both metabolism and oxidation (which yields a mixture of multiple reaction products), so the measured concentrations of EGCG plus gallic acid represent an estimate of EGCG absorption into the tissue. Furthennore. the gel was allowed to form around suppository samples without agitation (which may occur during in vivo use due to movements of tissue in the region of the cervix); this was done so that suppository erosion and gel formation was primarily determined by dissolution properties of the different formulas Flux was measured over a period of up to 24 hours after application of the fonnulations to the bovine vaginal tissue. The applied formulations comprised of gel produced by each of the 6 formulations described in Example 12 after incubation with a volume of PBS equivalent to the weight of each suppository sample for 24 hours at 37 C. Hie gel material was separated from undissolved solid suppository material and 20 microliter aliquots were applied to the mucosal tissue samples in Franz diffusion cells and spread evenly onto the exposed tissue area of 0.55 cm2on each cell.

[0614] At the end of 24 hours, the tissue was tape stripped three times and the tape strips discarded. The remaining tissue was then split into an outer tissue (i.e. the uppermost tissue layers exposed to the formulation, including the epithelium) and an inner tissue (which consisted of the underlying tissue below the outer tissue layer). The actives were then extracted from each tissue compartment in 3 ml methanol for 24 hours and the concentrations of the actives in the receptor fluid and tissue compartments were measured via LCMS detection.

[0615] The accumulated dose of quinine in the receptor fluid and in the outer and inner tissue are shown in Table 22 below.

[0616] Table 22: Delivered dose in pg / cm2of quinine into and through the bovine vaginal tissue. The accumulated dose of EGCG + gallic acid in the receptor fluid and in the outer and inner tissue are shown in Table 23 below. Table 23: Measured amount of EGCG + gallic acid in pg / cm2into and through the tissue.

[0617] Dose proportionality of drag delivery is a favorable property for formulations, enabling control of dosages and indicating that the actives are not bound or precipitated by interaction with excipients. Formulation 4 displayed proportionality of amounts of drug delivered first into the HPMC gel during its formation after exposure to moisture as shown in Example 12 and then in amounts of EGCG and quinine delivered into mucosal tissue.

Claims

CLAIMSWhat is claimed is:

1. A pharmaceutical composition comprising epigallocatechin gallate (EGCG), which lacks an EGCG degradant, or which comprises the EGCG degradant at a level less than 50%, 40%, 30%, 20%. 10%, 5%, 3%, 2%, or 1% of the EGCG, wherein the pharmaceutical composition is formulated as a suppository or atopical ointment, cream, or gel.

2. A pharmaceutical composition comprising epigallocatechin gallate (EGCG), in a dosage form comprising 25-75 mg, 30-70 mg, 40-60 mg, or 45-55 mg, e.g., 50 mg of the EGCG.

3. A pharmaceutical composition comprising EGCG, wherein the pharmaceutical composition lacks an excipient with antioxidant or reducing activity, or comprises an excipient with antioxidant or reducing activity at a level of less than 5, 4, 3, 2, or 1 mg per dosage form (e.g., less than 2.5, 2, 1.5, 1, or 0.5 mg of excipient with antioxidant or reducing activity / g of suppository).

4. A pharmaceutical composition comprising EGCG, wherein the pharmaceutical composition lacks a reducing agent other than EGCG, or comprises a reducing agent other than EGCG at a level of less than5. 4, 3, 2, or 1 mg per dosage form (e.g., less than 2.5, 2, 1.5, 1, or 0.5 mg / ml).

5. The pharmaceutical composition of any of the preceding claims, which further comprises:(i) quinine or a salt thereof, (e.g., quinine hydrochloride, quinine sulfate, or quinine sulfate dihydrate), quinine congeners and metabolites, quinidine, synthetic quinoline compounds, chloroquine, hydroxychloroquine, mefloquine, primaquine, quinacrine, and / or acridine orange;(ii) zinc or a salt thereof, e.g., zinc acetate, zinc gluconate, or zinc citrate;(iii) sucrose or a salt thereof; and / or(iv) citric acid (e.g., citric acid monohydrate) or a salt thereof.

6. The pharmaceutical composition of claim 5. wherein:(i) the EGCG is present at an equal amount by weight as the quinine or salt thereof;(ii) the EGCG is present at a higher amount by weight than the zinc or salt thereof;(iii) the quinine or salt thereof is present at a higher amount by weight than the zinc or salt thereof; and / or(iv) the ratio of the EGCG to quinine or salt thereof to zinc or salt thereof is 10 + / - 20%: 10 + / - 20% : 1 + / - 20%.

7. The pharmaceutical composition of claim 5 or 6, wherein:(i) the EGCG is present at 1-10%, e.g., 1.5-8.5%, e.g.. 2-6%, e.g., 2.5-5%, e.g., about 2.5% or about 5% w / w;(ii) the quinine or the salt thereof is present at 1-10%, e.g., 1.5-8.5%, e.g., 2-6%, e.g., 2.5-5%, e.g., about 2.5% or about 5%w / w;(iii) the citric acid (e.g., citric acid monohydrate) is present at 0.1-1%, e.g., 0.2-0.8%, e.g., 0.3- 0.7%, e.g., 0.4-0.6%, e.g., about 0.5% w / w;(iv) the sucrose is present at 1-4%, e.g., 1.5-3.5%. e.g., 2-3%, e.g., about 2.5% w / w: and / or(v) the zinc (e.g., zinc acetate) is present at 0.01-1%, e.g., 0.1-0.9%. e.g., 0. 1-0.5%, e.g., about 0.2%, e.g., 0.24% w / w.

8. Tire pharmaceutical composition of any of the preceding claims, comprising one or more (e.g., 1, 2, 3, 4, or 5) excipients, wherein the excipients comprise PEG300. PEG300 stearate (e.g., PEG300 monostearate), PEG400. PEG400 stearate (e.g., PEG400 monostearate). PEG1500, PEG1500 stearate (e.g., PEG1500 monostearate), PEG3350. PEG3350 stearate (e.g.. PEG3350 monostearate) and / or HPMC (e.g., HPMC K100M).

9. Tire pharmaceutical composition of any one of claims 5-8, comprising:(a) 1-10%, e.g., 1.5-8.5%, e.g., 2-6%, e.g., 2.5-5%, e.g., about 2.5% or about 5% w / w quinine or salt thereof, e.g., quinine sulfate dihydrate;(b) 1-10%, e.g., 1.5-8.5%. e.g., 2-6%. e.g., 2.5-5%, e.g.. about 2.5% or about 5% w / w EGCG:(c) 12.5-27.5%, e.g., 15-25%, e.g., 17.5-22.5%, e.g., about 20% w / w PEG 400 monostearate;(d)(i) 15-35%, e.g., 20-30%, e.g., 22.5-27.5%, e.g., about 25%, e.g., 25.16% w / w PEG 400; or(ii) a composition comprising PEG stearate, e.g., PEG300 monostearate and PEG1500 monostearate, e.g., 0.01-99.9% PEG300 monostearate and 0.01-99.9% PEG1500 monostearate, e.g., Tefose® 1500;(e) 30-50%, e.g., 35-45%, e.g., 37.5-42.5%, e.g., about 40% w / w PEG 3350;(f) 0.1-1%, e.g., 0.2-0.8%, e.g., 0.3-0.7%, e.g., 0.4-0.6%, e.g., about 0.5% w / w citric acid, e.g., citric acid monohydrate;(g) 1-4%, e.g., 1.5-3.5%, e.g., 2-3%, e.g., about 2.5% w / w sucrose;(h) 0.01-1%, e.g., 0.1-0.9%, e.g., 0.1-0.5%, e.g., about 0.2%, e.g., 0.24% w / w zinc acetate; and(i) 2.5-12.5%, e.g., 5-10%, e.g., 6-9%, e.g., about 7%, e.g., 6.6% w / w HPMC, e.g., HPMCK100M.

10. The pharmaceutical composition of any of the preceding claims, which is free of, is substantially free of, or comprises less than:(i) 75%, 50%, 25%, 10%, 1%, 0.1%, or 0.01% by weight of Sanguinaria canadensis (bloodroot); and / or(ii) 1%, 0.1%, or 0.01% by weight of Hydrastis, e g., Hydrastis tincture, thuja oil, tea tree oil, bitter orange oil. vitamins (e.g., vitamin A), iron, ferric sulfate, ferrous sulfate, green tea extract, catechins other than EGCG (e.g., (-)-epicatechin (EC), (-)-epicatechin-3-gallate (ECG), (-)-epigallocatechin (EGC). gallocatechin, catechin gallate, gallocatechin gallate), and / or theasinensin A.

11. Tire pharmaceutical composition, method, use, or composition for use of any of tire preceding claims, wherein the pharmacal composition is fonnulated as a suppository.

12. The pharmaceutical composition, method, use, or composition for use of claim 1 or 11, wherein the pharmaceutical composition, suppository, or topical ointment, cream, or gel is formulated for:(i) vaginal formulation;(ii) anal administration;(iii) topical administration; or(iv) oral administration.

13. A method of treating or preventing a cancer, pre-cancer, or infection in a subject in need thereof, comprising administering the pharmaceutical composition of any of the previous claims to a subject.

14. A pharmaceutical composition comprising quinine or a salt thereof, wherein the pharmaceutical composition is fonnulated as a suppository, and wherein the pharmaceutical composition: a) is free of. is substantially free of, or comprises less than 16%. 10%. 5%, 1%, 0.1%. or 0.01% by weight of, purified leaf extract of Neem; b) is free of, is substantially free of, or comprises less than 8%, 4%, 2%, 1%, 0.1%, or 0.01% by weight of, purified saponins:c) is free of is substantially free of or comprises less than 8%, 4%, 2%, 1%, 0. 1%, or 0.01% by weight of purified saponins from Sapindus muckerossi; and / or d) is free of is substantially free of or comprises less than 4%, 2%, 1%, 0.1%, or 0.01% by weight of Mentha citrata oil.

15. A method of treating a mucosal epithelial cancer (e.g., vulvar, vaginal, anal, or oropharyngeal malignancy), an epithelial neoplasia (e.g., cervical dysplasia, genital mucosal neoplasia, oral mucosal neoplasia, oropharyngeal dysplasia, oropharyngeal neoplasia, or anal mucosal neoplasia), cutaneous precancerous and cancerous lesions (e.g., actinic keratosis), a squamous cell cancer, warts (e.g., genital warts), cervical cancer, or HPV infection, in a subject in need thereof, comprising administering to the subject an effective amount of a pharmaceutical composition comprising zinc or a salt thereof, wherein the method does not comprise administration of or does not comprise administration of a substantial amount of (e.g., the composition is free of, or substantially free of, e.g., comprises less than 1%, 0.1%, or 0.01% by weight of each of): a) Sanguinaria canadensis (bloodroot); or b) bromelain.

16. A pharmaceutical composition comprising:(i) EGCG and quinine or a salt thereof(ii) EGCG and zinc or a salt thereof(iii) zinc or a salt thereof and quinine or a salt thereof(iv) EGCG, quinine or a salt thereof, and zinc and a salt thereof or(v) EGCG complexed with sucrose.

17. A pharmaceutical composition comprising: a first poly-ethylene glycol (PEG) having an average molecular weight between 300-500 g / mol (e.g., PEG400, e.g., PEG400 monostearate), a second PEG having an average molecular weight between 1000-2000 g / mol, (e.g., PEG1500); and a third PEG having an average molecular weight between 3000-5000 g / mol, (e.g.. PEG 3350 or PEG4000).

18. A pharmaceutical composition comprising:a first poly-ethylene glycol (PEG) having an average molecular weight between 1000-2000 g / mol, (e.g., PEG1500); and a second PEG having an average molecular weight between 3000-5000 g / mol, (e.g., PEG 3350 or PEG4000).

19. A pharmaceutical composition comprising: one or more PEGs (e.g., a PEG400, a PEG1500, and a PEG4000); and a hard fat comprising two or more of (e.g., all of): i) a mono-glyceride ester of a fatty acid; ii) a di-glyceride ester of a fatty acid; iii) a triglyceride ester of a fatty acid;(iv) one or more PEG stearates, e.g., one or more PEG monostearates comprising a PEG having a molecular weight in the range of 200-2000, e.g., one or more PEG monostearates comprising a PEG having a molecular weight in the range of 300-1500; and wherein optionally the fatty acids have a carbon chain length of CIO to Cl 8.

20. A pharmaceutical composition comprising: one or more PEG (e.g., a PEG400, a PEG1500, a PEG 3350, and / or a PEG4000); and hydroxypropyl methylcellulose (HPMC) (e.g., HPMC K100M).

21. A pharmaceutical composition comprising: epigallocatechin gallate (EGCG) or a salt thereof; and sucrose or a salt thereof, wherein the pharmaceutical composition is formulated as a suppository.

22. A pharmaceutical composition comprising: a first poly-ethylene glycol (PEG) having an average molecular weight between 300-500 g / mol (e.g., PEG400), a second PEG having an average molecular weight between 2850-3350 g / mol, (e.g., PEG3350); a hard fat comprising:(a) i) a mono-glyceride ester of a fatty acid; ii) a di-glyceride ester of a fatty acid; and iii) a triglyceride ester of a fatty7acid; and / or(b) one or more PEG stearates, e.g., one or more PEG monostearates comprising a PEG having a molecular weight in the range of 200-2000, e.g., one or more PEG monostearates comprising a PEG having a molecular weight in the range of 300-1500; hydroxypropyl methylcellulose (HPMC); and citric acid or a salt thereof.

23. A method of making or manufacturing a pharmaceutical composition, the method comprising combining two or more of (e.g., all of): epigallocatechin gallate (EGCG); quinine or a salt thereof; and zinc or a salt thereof (e.g.. zinc acetate, zinc gluconate, or zinc citrate).

24. A method of making or manufacturing a pharmaceutical composition, the method comprising combining two or more of (e.g., all of): one or more PEGs; a hard fat comprising:(a) i) a mono-glyceride ester of a fatty acid; ii) a di-glyceride ester of a fatty acid; and iii) a triglyceride ester of a fatty acid; and / or(b) one or more PEG stearates, e g., PEG monostearates comprising a PEG having a molecular weight in the range of 200-2000, e.g., PEG monostearates comprising a PEG having a molecular weight in tire range of 300-1500; hydroxypropyl methylcellulose (HPMC); and citric acid or a salt thereof.

25. A method of making or manufacturing a pharmaceutical composition, the method comprising combining: one or more PEGs; a hard fat comprising:(a) i) a mono-glyceride ester of a fatty acid; ii) a di-glyceride ester of a fatty acid; and iii) a triglyceride ester of a fatty acid; and / or(b) one or more PEG stearates, e.g., PEG monostearates comprising a PEG having a molecular weight in the range of 200-2000, e.g., PEG monostearates comprising a PEG having a molecular weight in tire range of 300-1500; epigallocatechin gallate (EGCG);quinine or a salt thereof (e.g., quinine sulfate dihydrate); and zinc or a salt thereof (e.g., zinc acetate, zinc gluconate, or zinc citrate).

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