Compositions and dosage forms for the treatment of HPV infections and HPV-induced neoplasms

Abstract

Pharmaceutically acceptable salts of acyclic nucleotide phosphoramidates, and pharmaceutical compositions, morphic forms and dosage forms thereof, for treating HPV and related conditions, including neoplasia.

Description

[Technical Field]

[0001] [CROSS-REFERENCE TO RELATED APPLICATIONS] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 391,283, filed July 21, 2022, U.S. Provisional Patent Application No. 63 / 400,661, filed August 24, 2022, Chinese Patent Application No. 202211206517.7, filed September 30, 2022, and U.S. Provisional Patent Application No. 63 / 412,413, filed September 30, 2022, which applications are incorporated herein by reference for all purposes.

[0002] The present invention provides compositions, advantageous salts, prodrugs, stereoisomers, morphic forms, dosage forms, and uses thereof for treating human papillomavirus (HPV) infection or related disorders such as HPV-induced neoplasia in a host in need thereof. [Background technology]

[0003] There is no direct treatment for human papillomavirus, which infected more than 43 million people in the US in 2018, with more than 13 million new cases, according to the Centers for Disease Control and Prevention.

[0004] Current treatment options for HPV infections are supportive and limited. Each has significant drawbacks. Commonly used drug treatments include salicylic acid, trichloroacetic acid, imiquimod, and podofilox. Both trichloroacetic acid and salicylic acid chemically burn wart tissue as a means of clearing the virus, frequently causing skin irritation, soreness, and pain in the process. Furthermore, salicylic acid is not used to treat HPV infections in the anogenital area. Imiquimod (Aldara™, Zyclara™) stimulates the immune system to clear the infection through toll-like receptor signaling, causing redness and swelling. Podofilox (Condylox™) destabilizes microtubules, disrupting tissue and preventing host cell replication.

[0005] Even more troublesome are HPV infections that cause cellular changes in human patients that have not yet progressed to cancer but have reached the neoplastic stage. HPV-induced neoplastic forms include cervical intraepithelial neoplasia ("CIN"), anal intraepithelial neoplasia ("AIN"), perianal intraepithelial neoplasia ("PAIN"), vulvar intraepithelial neoplasia ("VIN"), penile intraepithelial neoplasia (PIN), and vaginal intraepithelial neoplasia (VAIN). HPV-induced cancers include cervical cancer, anal cancer, perianal cancer, penile cancer, vaginal cancer, vulvar cancer, and oropharyngeal cancer.

[0006] It is crucial to identify and treat HPV-induced neoplasia before it progresses to potentially untreatable cancer. Nearly all cases of cervical cancer are caused by infection with oncogenic types of HPV, which are sexually transmitted. The primary goal of early screening, such as the Papanicolaou test (Pap smear), is to identify abnormal cervical cells that are highly cellularly altered so that they can be removed or destroyed.

[0007] Cervical intraepithelial neoplasia is often treated by observation (a wait-and-see approach) or excision or ablation of the cervical transition zone. Techniques include cryotherapy, laser therapy, loop electrosurgical procedure (LEEP), and cone biopsy. All of these surgical procedures injure the affected area and can leave scars. LEEP, the most common intervention, is effective in 60%–90% of cases but is associated with a significantly increased risk of miscarriage, ectopic pregnancy, and negative psychological outcomes. Despite extensive research, no medications have been approved to replace or be used in conjunction with these surgical methods.

[0008] Papillomaviruses are a group of non-enveloped DNA viruses that infect keratinized cells of the skin and mucous membranes, including the anogenital area, in humans. Papillomaviruses are known to cause cutaneous warts, genital warts, respiratory papillomatosis, and cancer. Several species of the alpha-papillomavirus genus contain high-risk HPV types that are likely to lead to human neoplasia and subsequent cancer. Most cancer-causing HPV types are caused by alpha-7 and alpha-9, including types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 73, and 82. The most common cancer-causing HPV types are types 16 and 18. HPV-16 and HPV-18 cause the majority of cervical cancers. The majority of venereal warts are caused by low-risk HPV types 6 and 11. Vaccines against HPV 6, 11, 16, and 18 are under development and may be effective if administered before sexual debut. However, HPV vaccines may offer little benefit to sexually active women who have already been infected with HPV.

[0009] Specific available prophylactic vaccines include Gardasil 9 (HPV 9-valent vaccine; HPV 6, 11, 16, 18, 31, 33, 45, 52, and 58), Gardasil 4 (4-valent), and Cervarix (2-valent). These are useful if the person is vaccinated before exposure to the virus, which typically means before sexual activity. Prophylactic vaccines are designed to produce neutralizing antibodies that eliminate the virus before it can infect cells. In contrast, therapeutic vaccines are vaccines that mount CD4+ and / or CD8+ T cell-based responses and are designed to eliminate HPV-infected cells. Exemplary antigens for therapeutic vaccines include E6 and E7. There are currently no approved therapeutic vaccines. Non-limiting examples of therapeutic vaccines being studied in clinical trials include VGX-3100 (INOVIO), GGX-188E (Genexine, Inc.), and ADXS11-001 (Advaxis, Inc.).

[0010] Cervical intraepithelial neoplasia (CIN) is a precursor to cervical cancer. As many as 20% of women infected with HPV have CIN (Non-Patent Document 1). CIN is graded on the Bethesda scale, ranging from mild grade 1 to severe grade 3. Typically, when a woman is diagnosed with grade 1 CIN, a "watch and wait" approach is taken. Due to the side effects of surgical approaches, treatment is only recommended if CIN is grade 2-3.

[0011] The cervical epithelium is composed of several layers of tissue, called stratified squamous epithelium. These layers are the superficial, intermediate, parabasal, and basal cell layers. It is essential that topical agents for the treatment of cervical intraepithelial neoplasia penetrate these multiple tissue layers to adequately reach and treat transformed cells. This presents a formidable challenge because the cells are tightly bound and lack blood vessels.

[0012] In 1996, a National Cancer Institute consensus panel pointed out the need for non-surgical intervention for cervical intraepithelial neoplasia (Non-Patent Document 2). Since the publication of that guidance, various approaches have been explored for the treatment of HPV and CIN, including immunomodulatory agents, antiproliferative agents, antiviral agents, and hormonal agents. However, there are no FDA-approved treatment options for HPV infection or CIN that have been proven effective in clinical trials (Non-Patent Document 3).

[0013] The Regents of the University of California have filed a series of patent applications with Karl Hostetler, et. al. as inventors, relating to various acyclic nucleotide derivatives for treating papilloma infections, including (i) U.S. Patent Nos. 6,111, 6,112, 6,113, 6,114, 6,115, 6,116, 6,117, 6,118, 6,119 ...

[0014] Antiva Biosciences conducted human clinical trials with the phosphonate ABI-1968 to evaluate its ability to sufficiently penetrate various layers of the cervical epithelium and release the antiviral agent PMEG ((9-[2-phosphonomethoxy)ethyl)guanine]). PMEG is then phosphorylated to the active compound, PMEGpp (PMEG polyphosphate). ABI-1968 was deemed unsuitable for topical use to treat cervical intraepithelial neoplasia because tissue concentrations of ABI-1968 did not reach 15 ng / mg, even at doses up to 3% (see bars F and G in Figure 116). Topically administering a drug to stratified HPV-infected epithelial tissue in an effective manner to destroy neoplastic cells in multiple epithelial layers presents significant challenges. The drug must be sufficiently lipophilic to penetrate tissue layers at sufficient concentrations to kill pathogenic cells and be metabolized to the active agent as needed. [ka]

[0015] Articles have been published discussing various topical drug delivery strategies, including semisolid dosage forms, gels, tablets, films, and pessaries. See, for example, J. M. Bach, J. Med. ...

[0016] It is an object of the present invention to provide effective pharmaceutical compositions and treatments for HPV infection and related conditions such as HPV-induced neoplasia, including but not limited to cervical intraepithelial neoplasia (CIN), anal intraepithelial neoplasia (AIN), vulvar intraepithelial neoplasia (VIN), penile intraepithelial neoplasia (PIN), perianal intraepithelial neoplasia (PAIN), and vaginal intraepithelial neoplasia (VAIN), in a host in need thereof. [Prior art documents] [Patent documents]

[0017] [Patent Document 1] U.S. Patent No. 8,835,603 [Patent Document 2] U.S. Patent No. 9,629,860 [Patent Document 3] U.S. Patent No. 9,156,867 [Patent Document 4] U.S. Patent No. 10,449,207 [Patent Document 5] U.S. Patent No. 10,195,222 [Patent Document 6] U.S. Patent No. 10,076,533 [Patent Document 7] U.S. Patent No. 10,076,532 [Patent Document 8] U.S. Patent No. 9,775,852 [Patent Document 9] U.S. Patent No. 9,387,217 [Patent Document 10] U.S. Patent No. 10,702,532 [Patent Document 11] U.S. Patent No. 10,213,430 [Patent Document 12] U.S. Patent No. 9,493,493 [Patent Document 13] U.S. Patent No. 9,801,884 [Patent Document 14] U.S. Patent No. 11,014,950 [Patent Document 15] U.S. Patent No. 10,377,782 [Non-patent literature]

[0018] [Non-Patent Document 1] Rozendaal, L. et al. "PCR-based high-risk HPV test in cervical cancer screening gives objective risk assessment of women with cytomorphologically normal cervical smears" 1996, Int J Cancer, 68, 766-769 [Non-patent document 2] National Institutes of Health Consensus Development Conference statement on cervical cancer. April 1-3, 1996. J Women's Health, 1996, 1, 1-38 [Non-patent document 3] Desravines, N. et al. "Topical therapies for the treatment of cervical intraepithelial neoplasia (CIN) 2-3: A narrative review" Gynecol Oncol Rep. 2020, 33, 100608 [Non-patent document 4] Keshari Sahoo, C. et al. "Intra vaginal Drug Delivery System: An Overview", 2013, American Journal of Advanced Drug Delivery, 1, 43-55 [Non-patent document 5] da Neves, J. et al. "Gels as vaginal drug delivery systems", 2006, International Journal of Pharmaceutics, 318 (2) 1-14 [Non-patent document 6] Cencia Rohan, L. et al. "Vaginal Drug Delivery Systems for HIV Prevention", 2009, AAPS, 11 (78) [Non-Patent Document 7] Kast, CE et al. "Design and in vitro evaluation of a novel bioadhesive vaginal drug delivery system for clotrimazole" Journal of Controlled Release, 2002, 81 (3) 347-354 [Non-patent document 8] Acarturk, F. "Mucoadhesive vaginal drug delivery systems", Recent Pat Drug Deliv Formula., 2009, 3 (3) 193-205 [Non-Patent Document 9] Sonal, G. et al. "Exploring Novel Approaches to Vaginal Drug Delivery", Recent Patents on Drug Delivery and Formulation, 2011, 5 (2) 82-94 Summary of the Invention

[0019] It has been discovered that an effective composition for treating HPV-infected and related diseases, including HPV-induced neoplasia, such as cervical intraepithelial neoplasia, anal intraepithelial neoplasia, perianal intraepithelial neoplasia, penile intraepithelial neoplasia, vulvar intraepithelial neoplasia, and vaginal intraepithelial neoplasia, requires the combination of several selected aspects working together to achieve the desired result. To achieve the desired ability to penetrate epithelial layered tissue in effective amounts to deliver the active agent, it was essential to select an appropriate compound with favorable lipophilicity and tissue penetration properties, in conjunction with a selected pharmaceutically acceptable salt (optionally in an advantageous morphic form). After many failures, it took years of research to solve this problem for the benefit of patients worldwide who suffer from intraepithelial neoplasia, which can potentially become cancerous.

[0020] Specifically, the primary compounds were found to be the following specific salts: [ka]

[0021] Compound I is (ethyl((2-(2-amino-6-methoxy-9H-purin-9-yl)ethoxy)methyl)(benzyloxy)-phosphoryl)-L-alaninate. U.S. Patent No. 11,344,555, assigned to the Regents of the University of California, generally claims Compound I and pharmaceutically acceptable salts, and methods of using them in the treatment of papillomavirus infections. Compound I is an acyclic nucleotide phosphonate that is metabolized to a known, potent antiviral compound (PMEG; ((9-[2-phosphonomethoxy)ethyl)guanine]), but PMEG has poor cell permeability and use-limiting systemic toxicity. The assignees have discovered ways to improve the local delivery of prodrugs in a manner that is rapidly taken up by epithelial cells, a difficult task to date, and one that ABI-1968 failed to accomplish.

[0022] Compound I (ethyl(-((2-(2-amino-6-methoxy-9H-purin-9-yl)ethoxy)methyl)-(benzyloxy)phosphoryl)-L-alaninate) has two asymmetric centers, one at the phosphorus atom and one at the amino acid moiety, either of which can be in the R or S configuration. Thus, compound I exists as four stereoisomers, i.e., two diastereomeric pairs: (R P ,S C ) / (S P ,S C ) and (R P ,R C ) / (S P ,R C ). Although U.S. Patent No. 1,344,555 and U.S. Patent No. 1,344,555 generally describe Compound I, these patents do not address the potential stereochemistry of the phosphorus atom. The stereoisomer of Compound I having R-stereochemistry at the phosphorus and S-stereochemistry at the amino acid carbon has been discovered to have advantageous properties over the other three stereoisomers, as discussed further herein. [ka]

[0023] In a non-limiting embodiment, a preferred salt (e.g., fumarate salt) of Compound I is used as a mixture of (R,S) and (S,S) diastereomers, where the first R / S indicates the stereochemistry at the phosphorus atom and the second S indicates the stereochemistry of the carbon in the amino acid moiety (corresponding to an L-alanine residue with S configuration). Any ratio of diastereomers that produces the desired results can be used, but the (R,S) diastereomer is prominent. In other embodiments, the ratio of R to S enantiomers at the phosphorus atom is approximately 1:1. In aspects, the compound is enantiomerically enriched in the R chirality at the phosphorus atom, with the amount of R by weight being, for example, greater than about 50%, or about 60%, 70%, 75%, 80%, or even 85% or more.

[0024] The S-configuration of the asymmetric carbon, corresponding to the natural amino acid configuration, is advantageous in the present invention. In certain embodiments, the amount of S by weight is, for example, greater than about 50%, or about 60%, 70%, 75%, 80%, or even 85% or more. In alternative embodiments, compounds are used that have the R-configuration at the asymmetric carbon, and that are greater than about 50%, or about 60%, 70%, 75%, 80%, or even 85% or more of the R-configuration.

[0025] Enantiomerically pure (R p ,S c , or simply "R,S") versions are the primary embodiment. Unless otherwise noted, enantiomerically pure Compound II is at least 90% free of the opposite enantiomer. Surprisingly, this compound would not have been chosen as the active ingredient for a topical formulation because it is an oil rather than a solid. This is especially true because the racemic mixture or enantiomerically enriched R,S isomer of the S,S free base is a solid. Furthermore, the S,S isomer has moderate crystallinity, as seen in Figure 120. However, when formed as the fumarate salt, R,S enantiomerically pure Compound I becomes a highly crystalline material, making it most advantageous for intraepithelial topical administration. Thus, the mono-fumaric acid salt of Compound I exhibits unexpected stability and processability, making it therapeutically advantageous over the free base of Compound I.

[0026] The R,S isomer monofumarate can be easily crystallized from isopropanol and heptane. This morphic form is anhydrous with a melting point of about 140°C (Example 15). This morphic form has been reproduced on both milligram and multigram scales.

[0027] While the S,S isomer was more tractable as the free base, the monofumarate salt of the S,S isomer is polymorphic and has a low melting point of approximately 105°C. Four morphic forms of the S,S monofumarate salt were identified (Example 15). In certain experiments, dissociation of the S,S monofumarate to the hemifumarate was observed. This pattern of synthesis could not be reproduced on a larger scale.

[0028] Surprisingly, it has been discovered that certain pharmaceutical dosage forms prepared from Compound I monofumarate and its morphic form, Pattern 1, have advantageous properties. Tablets prepared from Compound I free base substantially degrade within one month at 40° C. and 75% RH, whereas tablets prepared from Compound I monofumarate exhibit much less degradation (Example 25), resulting in significantly improved shelf life.

[0029] Compound II, referred to herein and illustrated below, is an enantiomerically enriched or pure embodiment in which R-stereochemistry at the phosphorus atom and S-stereochemistry at the amino acid carbon atoms predominate. In its enantiomerically pure form, Compound II exhibits superior stability to its stereoisomer, ethyl ((S)-(2-(2-amino-6-methoxy-9H-purin-9-yl)ethoxy)methyl(benzyloxy)phosphoryl)-L-alaninate monofumarate (Compound III). This is important for successful topical application to the cervix, vagina, vulva, perianal region, anus, or penis. [ka]

[0030] Other advantageous salts of Compound I that have been discovered include the hemifumarate salts ethyl ((R)-(2-(2-amino-6-methoxy-9H-purin-9-yl)ethoxy)methyl(benzyloxy)phosphoryl)-L-alaninate hemifumarate (Compound IV) and ethyl ((S)-(2-(2-amino-6-methoxy-9H-purin-9-yl)ethoxy)methyl(benzyloxy)phosphoryl)-L-alaninate hemifumarate (Compound V). [ka]

[0031] Compound II has been found to have high tissue penetration, surprisingly stable crystals, and is non-hygroscopic. Compound II and its advantageous morphic form, Pattern 1, can be used to treat HPV infection or diseases associated with HPV infection, such as intraepithelial neoplasia, including but not limited to cervical intraepithelial neoplasia, anal intraepithelial neoplasia, vulvar intraepithelial neoplasia, penile intraepithelial neoplasia, perianal intraepithelial neoplasia, and vaginal intraepithelial neoplasia, and can prevent the progression to cancer.

[0032] There are many strains of HPV, some of which are known as high-risk strains because they are strongly associated with the development of cancer. Compound I fumarate or Compound II can be used to treat high-risk types of HPV, including HPV-16 and HPV-18. Thus, in certain embodiments, the present invention provides Compound II and isolated morphic forms of Compound II Pattern 1, pharmaceutical compositions containing such compounds, methods for treating HPV infection or intraepithelial neoplasia associated with HPV infection using selected morphic forms described herein, and methods for preparing such compounds and morphic forms.

[0033] In particular, it has been surprisingly discovered that the monofumarate salt of ethyl ((R)-(2-(2-amino-6-methoxy-9H-purin-9-yl)ethoxy)methyl)(benzyloxy)phosphoryl)-L-alaninate (Compound II) has very high tissue penetration when administered topically to target tissues. Local administration avoids the toxicity associated with systemic drug administration. Because HPV-infected precancerous and / or cancerous cells reside in several layers of the epithelium, compounds must have high tissue penetration to reach and treat these affected cells.

[0034] Compound I monofumarate has superior tissue permeability and penetration in both porcine and human vaginal tissues compared to ABI-1968, a phosphonate ester of an acyclic purine nucleoside that also failed in clinical trials. Compound I monofumarate reaches vaginal tissue concentrations of 40 ng / mL to 85 ng / mL at a 0.1% dose. ABI-1968 does not even reach a concentration of 15 ng / mL at a 3% dose (see Figure 116). This significant improvement in tissue penetration was unexpected, especially considering the reduced dose.

[0035] Compound II is the corresponding S P Compound II is surprisingly stable compared to its isomer (Compound III). As shown in Example 7, Table 9, Compound II has a melting point of about 140°C ± 10°C, e.g., 141.5°C, while Compound III has a melting point of about 100°C ± 10°C, e.g., 106.4°C. Compound II is also much more crystalline than Compound III, as evidenced by XRPD data comparing the mono-fumarate salts of both compounds (see Example 13, Table 37 and Figure 71 compared to Example 15, Table 40 and Figure 77).

[0036] Careful selection of each aspect of the present invention was essential to achieve the desired results. One important aspect is formulation. Topical formulations used in the present invention include semi-solid dosage forms such as gels, creams, ointments, liquid or solid dosage forms. Non-limiting examples of solid dosage forms include tablets that can be inserted into the affected area.

[0037] It has been discovered that Compound I monofumarate, Compound II, or Compound III can be prepared in solid dosage forms for topical administration. In some embodiments, tablet formulations provide similar tissue penetration to gel formulations (55 ng / mg to 85 ng / mg for gels, 44 ng / mg to 79 ng / mg for tablets; Figure 121).

[0038] High crystallinity facilitates the isolation and processing of pharmaceutical compounds. Compound II exhibits surprisingly low hygroscopicity compared to Compound III. When subjected to a 40%-0%-95%-0%-45% relative humidity cycle, Compound II retains approximately 0.25% water content and the XRPD pattern remains unchanged (Example 21, Table 46). When exposed to the same conditions, Compound III retains approximately 10% water content, a 40-fold increase. These conditions also result in the loss of one peak in the XRPD pattern, indicating that Compound III changes morphic form in response to humidity changes. The hygroscopicity and stability advantages of Compound II over Compound III are surprising and unpredictable.

[0039] Compound II Pattern 1 can be produced, for example, by recrystallizing Compound II (Example 13, Table 37) and equilibrating in an appropriate solvent (Example 22). In certain embodiments, Compound II can be dissolved in an alcoholic solvent (e.g., isopropanol) and crystallized as Pattern 1 by the addition of an aliphatic solvent (e.g., heptane). In certain embodiments, Compound II can be dissolved in an alcoholic solvent (e.g., ethanol) and crystallized as Pattern 1 by the addition of an aliphatic solvent (e.g., heptane). Compound II Pattern I can also be prepared by equilibration in isopropanol, heptane, water, acetone, isopropanol:heptane (3:10), isopropanol:MTBE (1:3), and ethyl acetate:toluene (1:3).

[0040] Compound III Pattern I can be prepared in multiple steps (see Example 15). First, the free base of Compound III was dissolved in isopropanol. One equivalent of fumaric acid was added to induce precipitation. After heptane was added, the mixture was stirred at an elevated temperature, e.g., 50°C, for 20 hours and then cooled. Another 0.2 equivalents of fumaric acid was added along with heptane, and the mixture was stirred at an elevated temperature for at least about 13 hours. The suspension was then slowly cooled to below about 5°C and stirred at that temperature for at least about 2 days. The resulting solid Compound III Pattern I was collected by filtration.

[0041] Compound I (i.e., a mixture of R and S enantiomers at the phosphorus atom and S stereoisomer at the amino acid carbon) monofumarate Pattern 1 can be produced, for example, by recrystallization of Compound I monofumarate (Example 12, Table 31), equilibration of Compound I monofumarate in a suitable solvent, or crystallization by slow evaporation of the solvent (Example 12, Table 32). In certain embodiments, Compound I monofumarate can be dissolved in an alcoholic solvent (e.g., isopropanol) and crystallized as Pattern 1 by the addition of an aliphatic solvent (e.g., heptane). In certain embodiments, Compound I monofumarate can be dissolved in an alcoholic solvent (e.g., isopropanol) and crystallized as Pattern 1 by the addition of an ethereal solvent (e.g., methyl tert-butyl ether). In certain embodiments, Compound I monofumarate Pattern 1 can be produced by equilibration in a mixture of an ethereal solvent (e.g., tetrahydrofuran) and an aliphatic solvent (e.g., heptane). In certain embodiments, compound I monofumarate pattern 1 can be produced by crystallization via slow evaporation of a solvent at room temperature. Suitable solvents for slow evaporation crystallization of compound I monofumarate pattern 1 include, but are not limited to, acetone, methyl ethyl ketone, ethyl acetate, methanol, ethanol, isopropanol, and tetrahydrofuran. In certain embodiments, compound I monofumarate pattern 1 is characterized by an XRPD pattern comprising at least three 2θ values selected from 6.0+0.2°, 8.9+0.2°, 9.6+0.2°, 11.1+0.2°, 11.9+0.2°, 14.8+0.2°, 15.3+0.2°, 18.1+0.2°, 20.2+0.2°, 23.1+0.2°, 25.2+0.2°, and 27.0+0.2° (see Example 7).

[0042] Other morphic forms of Compound I monofumarate have been prepared, including Pattern 2, Pattern 3, and Pattern 4. However, these morphic forms can be unstable and result in the hemifumarate (a mixture of Compound IV and Compound V) even when prepared from the monofumarate.

[0043] Recrystallization of Compound I monofumarate in methyl ethyl ketone, acetone, acetone and heptane, methyl ethyl ketone and heptane, and ethanol and methyl tert-butyl ether all yield the hemifumarate pattern 2. 1 The ratio of Compound I as the free base to the fumarate, as determined by H NMR, is about 1:0.5, e.g., 1:0.52. In one embodiment, Pattern 2 is characterized by an XRPD pattern comprising at least three 2-theta values selected from 4.3±0.2°, 6.2±0.2°, 9.0±0.2°, 13.0±0.2°, 17.7±0.2°, 18.7±0.2°, and 25.3±0.2° (see Example 12).

[0044] Recrystallization of compound I monofumarate in acetonitrile or acetonitrile and water affords the hemifumarate pattern 3. After isolation by filtration, 1 The ratio of Compound I as the free base to fumarate, as determined by H NMR, is approximately 1:0.95, but washing with water reduces this ratio to approximately 1:0.76. In one embodiment, Pattern 3 is characterized by an XRPD pattern comprising at least three 2-theta values selected from 3.5±0.2°, 5.1±0.2°, 6.2±0.2°, 6.9±0.2°, 10.2±0.2°, 15.3±0.2°, 17.6±0.2°, 21.2±0.2°, and 28.9±0.2° (see Example 12). Recrystallization of Compound I monofumarate in acetone and toluene affords Pattern 4, which is the hemifumarate. 1The ratio of Compound I as the free base to the fumarate, as determined by H NMR, is about 1:0.7, e.g., 1:0.69. In one embodiment, Pattern 4 is characterized by an XRPD pattern comprising at least three 2-theta values selected from 4.0±0.2°, 6.0±0.2°, 11.8±0.2°, 13.2±0.2°, 14.8±0.2°, 17.7±0.2°, 20.4±0.2°, and 25.2±0.2° (see Example 12). Due to the superior properties of the monofumarate relative to the hemifumarate, Compound I monofumarate Pattern 1 was selected for further study due to its surprising stability and crystallinity.

[0045] In exemplary, non-limiting embodiments, there is provided a method for treating HPV-induced intraepithelial neoplasia, comprising administering an effective amount of one or a combination of active compounds described herein in a topical formulation sufficient to treat the neoplasia.

[0046] In exemplary embodiments, the formulation for treating intraepithelial neoplasia is in a dosage form containing 0.005 mg to 50 mg, 0.05 mg to 40 mg, 0.1 mg to 30 mg, 0.5 mg to 20 mg, 1 mg to 20 mg, 1 mg to 15 mg, or 1 mg to 10 mg of Compound I monofumarate, Compound II, or Compound III.

[0047] In certain embodiments, a formulation for treating intraepithelial neoplasia is in a dosage form containing about 0.001 mg to about 20 mg, about 0.005 mg to about 10 mg, about 0.01 mg to about 5 mg, about 0.03 mg to about 1 mg, about 0.05 mg to about 0.3 mg, about 0.03 mg to about 0.07 mg, about 0.05 mg to about 0.15 mg, or about 0.15 mg to about 0.45 mg of Compound I monofumarate, Compound II, or Compound III.

[0048] In certain embodiments, the formulation for treating intraepithelial neoplasia is in a dosage form containing about 0.001 milligrams to about 0.005 milligrams, about 0.005 milligrams to about 0.01 milligrams, about 0.01 milligrams to about 0.03 milligrams, about 0.03 milligrams to about 0.25 milligrams, about 0.20 milligrams to about 0.5 milligrams, about 0.4 milligrams to about 1 milligram, about 0.75 milligrams to about 3 milligrams, about 1 milligram to about 10 milligrams, or about 5 milligrams to about 20 milligrams. In certain embodiments, the formulation for treating intraepithelial neoplasia is in a dosage form comprising about or at least 0.005 mg, 0.01 mg, 0.03 mg, 0.05 mg, 0.1 mg, 0.3 mg, 0.5 mg, 0.7 mg, 1 mg, 1.5 mg, 2 mg, 2.5 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, or 50 mg of Compound I monofumarate, Compound II, or Compound III.

[0049] Specific dosage amounts are 0.05 mg, 0.1 mg, 0.2 mg, or 0.3 mg of Compound I monofumarate, Compound II, or Compound III. In certain embodiments, 0.05 mg, 0.1 mg, 0.2 mg, or 0.3 mg dosages of Compound I monofumarate, Compound II, or Compound III are administered once, twice, or three times per week as needed. In certain embodiments, a 0.05 mg dosage of Compound I monofumarate, Compound II, or Compound III is administered for a predetermined time period as directed by a healthcare professional, including daily dosing.

[0050] In certain embodiments, the topical formulation is administered twice daily, once daily, or several days per week (e.g., two or three days per week) as needed to achieve the desired results. In certain embodiments, the topical formulation is administered on a weekly schedule for one, two, three, four, five, six, or more weeks. In certain aspects, the topical formulation is administered on a schedule of three administrations per week for two, three, four, five, or six weeks.

[0051] In certain embodiments, the compound can be administered in one or more treatment cycles, including a treatment cycle and a rest cycle, where a treatment cycle comprises administering the compound as described herein, followed by a rest cycle (including a period of no treatment) before the next treatment cycle. In certain embodiments, the rest cycle lasts from about 1 day to about 6 months. In certain embodiments, the rest cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, or more before the next treatment cycle. In certain embodiments, multiple treatment cycles are administered, for example, 1, 2, 3, 4, 5, or 6 treatment cycles.

[0052] Dosage forms that do not adhere well to the target site may be removed, which may interfere with treatment. Dosage forms that adhere to the target site and dissolve rapidly with low flow rates have been discovered. Adhesion to the target site may also prevent exposure to non-target tissues, limiting toxicity, unwanted systemic exposure, and side effects. Dosage forms that soften, degrade, and / or disintegrate quickly with low flow rates are advantageous for causing rapid release of the active compound to the target tissue. For example, dosage forms that disintegrate with less than about 50 μL, less than about 100 μL, less than about 125 μL, less than about 150 μL, less than about 175 μL, less than about 200 μL, less than about 250 μL, less than about 500 μL, less than about 1 mL, or less than about 2 mL of fluid facilitate penetration of the drug into the target site.

[0053] In certain embodiments, the dosage form is a semi-solid, such as a gel or cream. In certain embodiments, the dosage form is a tablet. In certain embodiments, the dosage form disintegrates in about 1 second to about 10 seconds. In certain embodiments, the dosage form disintegrates in about 10 seconds to 1 minute, and in certain embodiments, the dosage form disintegrates in about 1 minute to about 1 hour. In certain embodiments, the dosage form disintegrates in 1 hour to 6 hours.

[0054] The physical dimensions of the dosage form can affect its effectiveness. Thinner tablets have a higher surface area to volume ratio, which may lead to faster disintegration and better coverage of the target area. In certain embodiments, the dosage form is less than 3 millimeters thick in its smallest dimension.

[0055] The formulation of the dosage form is critical to proper administration of the active agent to intraepithelial tissues. For example, the formulation can be prepared for use as a tablet, a reconstituted powder, a dry powder, a semi-solid dosage form, a film, or a pessary (i.e., a vaginal suppository).

[0056] Tablet formulations should exhibit mucoadhesive and substantivity properties and should contain excipients with solubility, erosion (for disintegration), porosity (for water uptake), and viscosity (to retain the drug at the target site) properties.

[0057] Examples of excipients that cause rapid disintegration of solid dosage forms covering the cervix, anus, penis, perianal, vulva, or vaginal areas include, but are not limited to, mannitol, microcrystalline cellulose, lactose, sucrose, calcium phosphate, sodium phosphate, sodium bicarbonate, citric acid, maleic acid, adipic acid, or fumaric acid. Examples of excipients that can enhance disintegration and coverage of the affected area include, but are not limited to, sodium starch glycolate, pregelatinized starch, crospovidone, and croscarmellose sodium. Examples of mucoadhesive excipients useful in the present invention include, but are not limited to, microcrystalline cellulose, polycarbophil, hydroxymethylcellulose, hypromellose, hydroxypropylcellulose, and PVP.

[0058] A non-limiting example of a tablet formulation includes, but is not limited to, microcrystalline cellulose, crospovidone, magnesium stearate, silicon dioxide, polyethylene oxide, and mannitol. Another non-limiting example of a tablet formulation has microcrystalline cellulose, magnesium stearate, and mannitol.

[0059] Alternative formulations are reconstituted or dry powders. These formulations may include the excipients described above, and in certain embodiments, xanthan gum may be added. As a non-limiting example, a dry powder formulation may include, but is not limited to, xanthan gum, mannitol, silicon dioxide, and sodium benzoate.

[0060] The semi-solid dosage form may contain, for example, a mucoadhesive polymer, a solubility / penetration enhancer, a lipophilic solubilizer, and a penetration enhancer. The mucoadhesive polymer may be, for example, but not limited to, carbomer, polyethylene glycol, crospovidone, hypromellose, polycarbophil, and / or hydroxyethylcellulose. The solubility / penetration enhancer may be, for example, but not limited to, a mixture of polyoxyl 6 type I stearate, ethylene glycol stearate, and polyoxyl 32 type I stearate, cetyl alcohol, stearyl alcohol, polysorbate 80, sodium lauryl sulfate, monoglycerides and diglycerides, sorbitan monostearate, glyceryl isostearate, polyoxyl 15 hydroxystearate, polyoxyl 40 hydrogenated castor oil, octyldodecanol, and / or soybean lecithin. The lipophilic solubilizer may include, but is not limited to, light mineral oil, mineral oil, white wax, and silicone fluid. Penetration enhancers include, but are not limited to, propylene glycol, transcutol, oleic acid, isopropyl myristate, propylene glycol, glycerol monooleate, propylene glycol monocaprylate, PEG-8 beeswax, cetyl alcohol, stearic acid, cetyl palmitate, and / or cetostearyl alcohol.

[0061] A non-limiting example of a semi-solid formulation includes, for example, carbomer, propylene glycol, sorbic acid, EDTA, and water. Another non-limiting example of a semi-solid formulation includes carbomer, mineral oil, a mixture of polyoxyethylene 6 stearate type I, ethylene glycol stearate, and polyoxyethylene 32 stearate type I, paraben, propylene glycol, EDTA, and / or water.

[0062] Films can be made using, for example, but not limited to, hypromellose, polyethylene glycol, polymethacrylate, microcrystalline cellulose, xanthan gum, guar gum, and / or polyvinylpyrrolidone.

[0063] Pessaries (vaginal suppositories) can be formulated using, for example, but not limited to, hard fat (such as Ovucire, Witepsol™, Supposi-Base, etc.), polyethylene glycol, macrogol, cocoa butter, and glycerol. Non-limiting examples of pessaries can be made from Witepsol™ H 15 or Ovucire WL 3264.

[0064] Thus, the present invention includes at least the following features: (i) Compound I monofumarate; (ii) Compound II; (iii) Compound III; (iv) Compound IV; (v) Compound V; (vi) compounds of (i), (ii), (iii), (iv) or (v) in enantiomerically enriched or enantiomerically pure form; (vii) Compounds of (ii) wherein the amount of R by weight is, for example, greater than about 50%, or about 60%, 70%, 75%, 80%, or even 85% or more: (viii) compounds of (vii) in which the amount of S-configuration at the asymmetric carbon is greater than about 50%, or greater than about 60%, 70%, 75%, 80%, or even 85%; (ix) enantiomerically pure (R,S) compound I; (x) enantiomerically pure (S,S) compound I; (xi) R,S enantiomerically pure Compound II in highly crystalline form; (xii) Compound II Pattern 1; (xiii) morphic forms more particularly described in Section III; (xiv) a topical pharmaceutical composition comprising an effective amount of an active compound described herein or a morphic form thereof and a pharmaceutically acceptable carrier; (xv) an external preparation of (xiv) in the form of a tablet; (xvi) a tablet dosage form of (xv), comprising Compound I monofumarate, mannitol, polycrystalline cellulose, and magnesium stearate; (xvii) a tablet dosage form of (xv), comprising Compound II, mannitol, polycrystalline cellulose, and magnesium stearate; (xviii) an external preparation of (xiv) in the form of a semisolid dosage form; (xix) a semi-solid dosage form of (xviii), comprising Compound I monofumarate, light mineral oil, propylparaben, Tefose™ 63, water, EDTA, methylparaben, and Carbopol™ 974P; (xx) a semi-solid dosage form of (xviii), comprising Compound I monofumarate, water, EDTA, methylparaben, Carbopol™ 974P, propylene glycol, and optionally sorbic acid; (xxi) a semi-solid dosage form of (xviii), comprising Compound II, light mineral oil, propylparaben, Tefose™ 63, water, EDTA, methylparaben, and Carbopol™ 974P; (xxii) a semi-solid dosage form of (xviii), comprising Compound II, water, EDTA, methylparaben, Carbopol™ 974P, propylene glycol, and optionally sorbic acid; (xxiii) a topical formulation of (xiv) in the form of a reconstituted powder; (xxiv) an external preparation of (xiv) in the form of a dry powder dosage form; (xxv) an external preparation of (xiv) in the form of a film; (xxvi) an external preparation of (xiv) in the form of a pessary; (xxvii) (xv) to (xxvi) in dosage forms advantageous for delivery to the cervix, vagina, vulva, penis, perianal area, and / or anus; (xxviii) A method of treating HPV-induced infections or associated conditions, including but not limited to intraepithelial neoplasias, such as those of the cervix, vagina, vulva, perianal, anus, or penis, comprising administering to a host in need thereof an effective amount of a compound, morphic form, or pharmaceutical composition of any one of the above embodiments; (xxix) Use of any of the above embodiments in the manufacture of a medicament for the treatment of an HPV infection or an associated condition in a host in need thereof, including, but not limited to, intraepithelial neoplasia of the cervix, penis, vulva, perianal skin, anus, or vagina; (xxx) Embodiments (i) to (xxvii) for use in treating an HPV infection or an associated condition in a host in need thereof, including, but not limited to, intraepithelial neoplasia of the cervix, penis, vulva, perianal, anus, or vagina; (xxxi) Any one of the above embodiments, wherein the host is a human; (xxxii) any of the above topical formulations for the treatment of intraepithelial neoplasia, providing a dosage form comprising 0.005 mg to 50 mg, 0.05 mg to 40 mg, 0.1 mg to 300 mg, 0.05 mg to 0.3 mg, 0.5 mg to 20 mg, 1 mg to 20 mg, 1 mg to 15 mg, 1 mg to 10 mg of a compound of embodiments (i) to (v), and in certain embodiments, about or at least 0.005 mg, 0.01 mg, 0.03 mg, 0.05 mg, 0.1 mg, 0.3 mg, 0.5 mg, 0.7 mg, 1 mg, 1.5 mg, 2 mg, 2.5 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, or 50 mg of a compound of embodiments (i) to (v); (xxxiii) any of the above topical formulations for the treatment of intraepithelial neoplasia, providing a dosage form comprising about 0.001 mg to about 20 mg, about 0.005 mg to about 10 mg, about 0.01 mg to about 5 mg, about 0.03 mg to about 1 mg, about 0.05 mg to about 0.3 mg, about 0.03 mg to about 0.07 mg, about 0.05 mg to about 0.15 mg, or about 0.15 mg to about 0.45 mg of the compound of embodiments (i) to (v); (xxxiv) The topical formulation of embodiments (xiv)-(xxvii) administered twice daily, once daily, or several days per week (e.g., 2 or 3 days per week), or for as long as necessary to achieve the desired result; (xxxv) The topical formulation of embodiments (xiv)-(xxvii) administered on a weekly schedule for 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks or more; (xxxvi) The method of (xxviii), further comprising applying a lubricant to the dosage form or epithelial tissue prior to inserting the dosage form into the affected area; (xxxvii) The method of (xxxvi), wherein the lubricant is selected from water, a glycerol-based lubricant, and a hydroxyethyl cellulose-based lubricant; and (xxxviii) A process for preparing the topical formulation according to any one of embodiments (xiv) to (xxviii); (xxxix) A method for treating HPV-induced infections or associated conditions, including but not limited to intraepithelial neoplasias of the cervix, penis, vulva, perianal, anus, or vagina, etc., comprising administering to a host in need thereof an effective amount of embodiment (i)-(xxii) in combination with surgical treatment of the target tissue before, during, or after administration of a compound; (xl) the method of (xxxix), comprising performing a surgical treatment of the target tissue, followed by administering an effective amount of an embodiment (i)-(xxii) to a host in need thereof; (xli) The method of (xxxiv), comprising administering an effective amount of any of embodiments (i)-(xxii) to a host in need thereof, followed by surgical treatment of the target tissue; (xlii) the method of (xxxiv), comprising administering to a host in need thereof an effective amount of an embodiment (i)-(xxii) before, after, or about simultaneously with surgical treatment of the target tissue; (xliii) The method of embodiments (xxxiv)-(xxxvii), wherein the surgical treatment of the target tissue is resection; (xliv) The method of embodiments (xxxiv)-(xxxvii), wherein the surgical treatment of the target tissue is ablation; (xlv) The method of (xliii), wherein the ablation is a loop electrosurgical ablation procedure (LEEP); (xlvi) (xliii) method in which the resection is a large loop excision of the transition zone (LLETZ); (xlvii) the method of (xliii), wherein the excision is a knife cone biopsy; (xlviii) The method of (xliii), wherein the excision is laser conization; (xlix) the method of (xliv), wherein the ablation is laser ablation; and (l) The method of (xliv) wherein the cauterization is cryocautery. (li) a process for preparing Compound II as described herein, and (lii) A process for producing the morphic forms described herein. [Brief explanation of the drawings]

[0065] [Figure 1] FIG. 1 shows a comparison of XRPD diffractograms of Compound I Pattern 1 obtained in an equilibration experiment in heptane (Example 2, Table 2, experiment EQ10), an anti-solvent precipitation in an acetone / methyl tert butyl ether (MTBE) system (Example 2, Table 3, experiment AS3), and a scaled-up preparation of Compound I Pattern 1 described in Example 3. [Figure 2] 1 is a DSC thermogram of Compound I Pattern 1 obtained in an equilibration experiment in heptane (Example 2, Table 2, experiment EQ10). [Figure 3] 1 is a TGA thermogram of Compound I Pattern 1 obtained in an equilibration experiment in heptane (Example 2, Table 2, experiment EQ10). [Figure 4] FIG. 1 is an XRPD diffractogram of Compound I Pattern 1 prepared in Example 3. [Figure 5] 1 is a DSC thermogram of Compound I Pattern 1 prepared in Example 3. [Figure 6] 1 is a TGA thermogram of Compound I Pattern 1 prepared in Example 3. [Figure 7] FIG. 1 shows a comparison of XRPD diffractograms of fumaric acid pattern, compound IV pattern 1 (obtained in Example 7), compound IV pattern 1 in mixture with fumaric acid pattern (sample RC2-EA, obtained in Example 5, experiment RC2), and compound IV pattern 1 (Example 5, sample RC3-EA). [Figure 8] FIG. 1 shows a comparison of XRPD diffractograms of Compound I monosuccinate pattern 1 (Example 5, sample RC7-IPA) and succinic acid pattern. [Figure 9] FIG. 1 shows a comparison of the XRPD diffractograms of Compound I monosuccinate pattern 1 of sample RC7-EA (Example 5, Table 5) and Compound I monosuccinate pattern 1 of sample AS7-B (Example 5, Table 6). [Figure 10] FIG. 1 shows a comparison of XRPD diffractograms of fumaric acid pattern, compound I monofumarate pattern 1 (Example 5, Table 6, sample AS2-B), and compound IV pattern 1 (Example 5, sample AS3-B). [Figure 11] FIG. 1 shows a comparison of the XRPD diffractograms of Compound I monosuccinate pattern 1 (Example 5, sample AS7-B) and the free succinic acid pattern. [Figure 12] FIG. 1 is an XRPD diffractogram of Compound I hemifumarate pattern 1 (Example 6, sample RC13). [Figure 13] 1 is a DSC thermogram of Compound I hemifumarate Pattern 1 (Example 6, Sample RC13). [Figure 14] 1 is a TGA thermogram of Compound I hemifumarate Pattern 1 (Example 6, Sample RC13). [Figure 15] FIG. 1 is an XRPD diffractogram of Compound I mosuccinate pattern 1 (Example 6, sample RC14). [Figure 16] FIG. 1 is a DSC thermogram of Compound I mosuccinate pattern 1 (Example 6, sample RC14). [Figure 17] 1 is a TGA thermogram of Compound I mosuccinate pattern 1 (Example 6, sample RC14). [Figure 18] FIG. 1 shows a comparison of the XRPD diffractograms of Compound I monofumarate pattern 1 (Example 6, sample RC16) and Compound I hemifumarate pattern 1 (Example 6, sample RC-13). [Figure 19] FIG. 1 shows a comparison of the XRPD diffractograms of Compound I hemisuccinate pattern 1 (Example 6, sample RC17) and Compound I monosuccinate pattern 1 (Example 6, sample RC-14). [Figure 20] FIG. 1 is a DSC thermogram of Compound I hemisuccinate pattern 1 (Example 6, sample RC17). [Figure 21] FIG. 1 is a TGA thermogram of Compound I hemisuccinate Pattern 1 (Example 6, Sample RC17). [Figure 22] FIG. 1 shows a comparison of the XRPD diffractograms of Compound I monofumarate pattern 1 (Example 6, sample RC18) and Compound I monofumarate pattern 1 (Example 6, sample RC-16). [Figure 23] FIG. 1 shows a comparison of the XRPD diffractograms of Compound I hemifumarate pattern 1 and Compound I monofumarate pattern 1 (small scale preparation) obtained in Example 7. [Figure 24] 1 is a DSC thermogram of Compound I hemifumarate pattern 1 obtained in Example 7. [Figure 25] 1 is a TGA thermogram of Compound I hemifumarate pattern 1 obtained in Example 7. [Figure 26A] 1 is a DSC thermogram (recorded at a heating rate of 10° C. / min) of Compound I monofumarate Pattern 1 (small scale preparation, Example 7). [Figure 26B] 1 is a DSC thermogram (recorded at a heating rate of 2° C. / min) of Compound I monofumarate Pattern 1 (small scale preparation, Example 7). [Figure 26C] FIG. 1 shows a DSC cycle (DSC cycle: 0° C. to 150° C., 150° C. to 0° C., 0° C. to 250° C., 10° C. / min) of Compound I monofumarate pattern 1 of a small-scale preparation sample (Example 7). [Figure 27] 1 is a TGA thermogram of Compound I monofumarate pattern 1 of a small-scale preparation sample (Example 7). [Figure 28] FIG. 10 is a comparison of the XRPD diffractograms of Compound I monofumarate Pattern 1 obtained in the stability testing experiments described in Example 8 at 25° C. / 84% RH (2 days, open container), 25° C. / 92% RH (1 week, open container), 40° C. / 75% RH (1 week, open container), and 60° C. (1 week, sealed container) with the original sample of Compound I monofumarate Pattern 1 before testing. [Figure 29] FIG. 1 shows a dynamic vapor sorption (DVS) plot of Compound I Pattern 1 (Example 10) and DVS change in mass plot. [Figure 30] FIG. 1 shows a comparison of the XRPD diffractograms of compound I pattern 1 (obtained in Example 3) before and after the DVS study (Example 10). [Figure 31] FIG. 1 shows a dynamic vapor sorption (DVS) plot of Compound I hemifumarate pattern 1 (Example 10) and DVS change in mass plot. [Figure 32] FIG. 1 shows a comparison of XRPD diffractograms of Compound I hemifumarate pattern 1 (obtained in Example 7) before and after DVS studies (Example 10). [Figure 33] FIG. 1 shows a dynamic vapor sorption (DVS) plot of Compound I monofumarate pattern 1 (Example 10) and DVS change in mass plot. [Figure 34] FIG. 1 shows a comparison of the XRPD diffractograms of compound I monofumarate pattern 1 (obtained in Example 7) before and after a DVS study (Example 10). [Figure 35] FIG. 1 shows a comparison of XRPD diffractograms of Compound I monofumarate pattern 1 obtained in Example 11 (large scale) before and after heating to 106° C. [Figure 36A]1 is a DSC thermogram (recorded at a heating rate of 10° C. / min) of Compound I monofumarate Pattern 1 (Example 11). [Figure 36B] 1 is a DSC thermogram (recorded at a heating rate of 2° C. / min) of Compound I monofumarate Pattern 1 (large scale preparation, Example 11). [Figure 36C] 1 is a DSC thermogram of Compound I monofumarate pattern 1 (obtained in Example 11). [Figure 37] 1 is a TGA thermogram of Compound I monofumarate pattern 1 (obtained in Example 11). [Figure 38] FIG. 1 shows a comparison of the XRPD diffractograms of Compound I monofumarate pattern 1, hemifumarate pattern 2, hemifumarate pattern 3, and pattern 4 obtained from a 2-week equilibration experiment in water at 25° C. with the fumaric acid pattern (obtained in Example 12). [Figure 39] FIG. 1 shows a comparison of XRPD diffractograms of Compound I Pattern 4 (obtained in Example 12) obtained from equilibration experiments in water at 25° C. for 2 and 3 weeks. [Figure 40] 1 is a DSC thermogram of Compound I Pattern 4 (obtained in Example 12) obtained from a 2-week equilibration experiment in water at 25° C. [Figure 41] 1 is a DSC thermogram of Compound I Pattern 4 (obtained in Example 12) obtained from a 3-week equilibration experiment in water at 25° C. [Figure 42] 1 is a TGA thermogram of Compound I Pattern 4 (obtained in Example 12) obtained from a 2-week equilibration experiment in water at 25° C. [Figure 43] 1 is a TGA thermogram of Compound I Pattern 4 (obtained in Example 12) obtained from a 3-week equilibration experiment in water at 25° C. [Figure 44] FIG. 1 shows a comparison of the XRPD diffractograms of the fumaric acid pattern and Compound I hemifumarate pattern C (obtained in Example 12) obtained from equilibration experiments EQ2 (in acetonitrile) and EQ15 (in 2.9:97.1 v / v water / acetonitrile) at 25° C. for 3 weeks. [Figure 45] 1 is a DSC thermogram of Compound I hemifumarate Pattern C (obtained in Example 12) obtained from a two-week equilibration experiment in acetonitrile at 25° C. [Figure 46] 1 is a TGA thermogram of Compound I hemifumarate Pattern C (obtained in Example 12) obtained from a two-week equilibration experiment in acetonitrile at 25° C. [Figure 47] FIG. 1 shows a comparison of XRPD diffractograms of Compound I hemifumarate pattern 2 (obtained in Example 12) obtained by equilibration experiments EQ3 (in methyl ethyl ketone), EQ4 (in acetone), and EQ7 (in 1:1 v / v acetone / heptane) at 25° C. for 3 weeks. [Figure 48] 1 is a DSC thermogram of Compound I hemifumarate Pattern 2 (obtained in Example 12) obtained from a two-week equilibration experiment in methyl ethyl ketone at 25° C. [Figure 49] 1 is a TGA thermogram of Compound I hemifumarate pattern 3 (obtained in Example 12) obtained from a two-week equilibration experiment in methyl ethyl ketone at 25° C. [Figure 50] FIG. 1 shows a comparison of XRPD diffractograms of a mixture of fumaric acid and Compound I monofumarate pattern 1 (obtained in Example 12) obtained in equilibration experiments EQ5 (in isopropanol), EQ8 (in 1:1 v / v isopropanol / heptane), EQ9 (in 1:1 v / v isopropanol / toluene), EQ10 (in 1:3 v / v isopropanol / methyl tert butyl ether), and EQ12 (in 1:3 v / v ethanol / heptane) at 25° C. for 3 weeks, and an unknown pattern. [Figure 51] 1 is a DSC thermogram of a mixture of Compound I monofumarate pattern 1 and an unknown pattern obtained in equilibration experiment EQ5 in isopropanol at 25° C. for 2 weeks (obtained in Example 12). [Figure 52] 1 is a TGA thermogram of a mixture of Compound I monofumarate pattern 1 and an unknown pattern obtained in equilibration experiment EQ5 in isopropanol at 25° C. for 2 weeks (obtained in Example 12). [Figure 53] FIG. 1 shows a comparison of XRPD diffractograms of compound I hemifumarate pattern 5 obtained in equilibration experiment EQ6 (in 1:1 v / v acetone / toluene) at 25° C. for 2 and 3 weeks (obtained in Example 12). [Figure 54] 1 is a DSC thermogram of Compound I hemifumarate pattern 5 obtained in equilibration experiment EQ6 (in 1:1 v / v acetone / toluene) at 25° C. for 2 weeks (obtained in Example 12). [Figure 55] 1 is a DSC thermogram of Compound I hemifumarate pattern 5 obtained in equilibration experiment EQ6 (in 1:1 v / v acetone / toluene) at 25° C. for 3 weeks (obtained in Example 12). [Figure 56] 1 is a TGA thermogram of Compound I hemifumarate pattern 5 obtained in equilibration experiment EQ6 (in 1:1 v / v acetone / toluene) at 25° C. for 2 weeks (obtained in Example 12). [Figure 57] 1 is a TGA thermogram of Compound I hemifumarate pattern 5 obtained in equilibration experiment EQ6 (in 1:1 v / v acetone / toluene) at 25° C. for 3 weeks (obtained in Example 12). [Figure 58] FIG. 1 shows a comparison of the XRPD diffractograms of Compound I monofumarate Pattern 1 obtained in a 2-week equilibration experiment at 25° C. (procedure of Example 12) EQ11 (in 1:3 v / v tetrahydrofuran / heptane), EQ13 (in 1:3 v / v ethyl acetate / toluene), EQ14 (in 1:3 v / v ethanol / toluene), and Compound I monofumarate Pattern 1 (material obtained in Example 7). [Figure 59] FIG. 1 shows a comparison of XRPD diffractograms of Compound I monofumarate pattern 1 (obtained in Example 12) obtained in equilibration experiments EQ11 (in 1:3 v / v tetrahydrofuran / heptane), EQ13 (in 1:3 v / v EA / toluene), and EQ14 (in 1:3 v / v ethanol / toluene) at 25° C. for 3 weeks. [Figure 60]Figure 1 shows a comparison of the XRPD diffractograms of the mixture of Compound I monofumarate pattern 1 and unknown pattern obtained in equilibration experiments EQ16 (in 1:4 v / v isopropanol / heptane), EQ5 (in isopropanol) at 25°C for 2 weeks, and Compound I monofumarate pattern 1 obtained in Example 7 (obtained in Example 12). [Figure 61] FIG. 1 shows a comparison of XRPD diffractograms of a reference fumaric acid pattern, Compound I monofumarate pattern 1 (Example 7), Compound I monofumarate pattern 2 (obtained by precipitation from an acetone solution with heptane anti-solvent in Experiment AS1, Example 12), Compound I monofumarate pattern 2 (obtained by precipitation from a methyl ethyl ketone solution with heptane anti-solvent in Experiment AS4, Example 12), and the fumaric acid pattern obtained in Experiment AS2, Example 12. [Figure 62] FIG. 1 shows a comparison of XRPD diffractograms of Compound I monofumarate pattern 1 (Example 7), Compound I monofumarate pattern 2 (Experiment AS6, obtained by precipitation from an ethanol solution with heptane anti-solvent in Example 12), and Compound I monofumarate pattern 2 (Experiment AS7, obtained by precipitation from a tetrahydrofuran solution with heptane anti-solvent in Example 12). [Figure 63] FIG. 1 shows a comparison of the XRPD diffractograms of Compound I monofumarate pattern 1 (Example 7) and Compound I monofumarate pattern 1 obtained by slow evaporation from acetone, methyl ethyl ketone, and ethyl acetate followed by crystallization at room temperature as described in Example 12, Table 32. [Figure 64] FIG. 1 shows a comparison of the XRPD diffractograms of Compound I monofumarate pattern 1 (Example 7) and Compound I monofumarate pattern 1 obtained by slow evaporation and crystallization at room temperature from methanol, ethanol, isopropanol, and tetrahydrofuran as described in Example 12, Table 32. [Figure 65]FIG. 1 shows a comparison of XRPD diffractograms of authentic fumaric acid pattern, Compound I monofumarate pattern 1 (Example 7), and Compound I monofumarate pattern 2 obtained by crystallization by slow cooling from a hot saturated solution of methyl ethyl ketone, Compound I monofumarate pattern 2 obtained by crystallization by slow cooling from a hot saturated solution of acetone, and Compound I monofumarate pattern 3 obtained by crystallization by slow cooling from a hot saturated solution of acetonitrile. [Figure 66] FIG. 1 shows a comparison of the XRPD diffractograms of Compound I monofumarate pattern 1 (Example 7), Compound I monofumarate pattern 1 obtained by crystallization by slow cooling from a hot saturated solution, and Compound I monofumarate pattern 1 obtained by crystallization by slow cooling from a hot saturated solution in ethanol / toluene (1 / 1 v / v) (Example 12, Table 33). [Figure 67] FIG. 1 shows a comparison of the XRPD diffractograms of Compound I monofumarate pattern 1 (Example 7), Compound I monofumarate pattern 2 obtained by crystallization by rapid cooling from a hot saturated solution in acetone, and Compound I monofumarate pattern 2 crystallized by rapid cooling from a hot saturated solution in methyl ethyl ketone (Example 12, Table 34). [Figure 68] FIG. 1 shows a comparison of XRPD diffractograms of authentic fumaric acid pattern, Compound I monofumarate pattern 1 (Example 7), Compound I monofumarate pattern 1 obtained by crystallization by rapid cooling from a hot saturated solution, Compound I monofumarate pattern 3 obtained by crystallization by rapid cooling from a hot saturated solution in acetonitrile, and Compound I monofumarate pattern 1 obtained by crystallization by rapid cooling from a hot saturated solution in ethanol / toluene (1 / 1 v / v) (Example 12). [Figure 69] FIG. 10 is a hot-cold DSC thermogram of Compound I monofumarate Pattern 1 (heated to 106° C.) (Example 12, Table 35). [Figure 70] FIG. 10 is a hot-cold DSC thermogram of Compound I monofumarate Pattern 1 (heated to 130° C.) (Example 12, Table 35). [Figure 71]FIG. 1 is an XRPD diffractogram of Compound II Pattern 1 (Example 13). [Figure 72] 1 is a DSC thermogram of Compound II Pattern 1 (Example 13). [Figure 73] 1 is a TGA thermogram of Compound II Pattern 1 (Example 13). [Figure 74] FIG. 1 is an XRPD diffractogram of compound IV pattern 1 (Example 14). [Figure 75] 1 is a DSC thermogram of Compound IV Pattern 1 (Example 14). [Figure 76] 1 is a TGA thermogram of Compound IV Pattern 1 (Example 14). [Figure 77] FIG. 1 is an XRPD diffractogram of Compound III Pattern 1 (Example 15). [Figure 78] 1 is a DSC thermogram of Compound III Pattern 1 (Example 15). [Figure 79] 1 is a TGA thermogram of Compound III Pattern 1 (Example 15). [Figure 80] FIG. 1 is an XRPD diffractogram of Compound III Pattern 2 (Example 16). [Figure 81] 1 is a DSC thermogram of Compound III Pattern 2 (Example 16). [Figure 82] 1 is a TGA thermogram of Compound III Pattern 2 (Example 16). [Figure 83] FIG. 1 is an XRPD diffractogram of Compound V Pattern 1 (Example 17). [Figure 84] 1 is a DSC thermogram of Compound V Pattern 1 (Example 17). [Figure 85] 1 is a TGA thermogram of Compound V Pattern 1 (Example 17). [Figure 86] FIG. 1 is an XRPD diffractogram of Compound V Pattern 2 (Example 18). [Figure 87] 1 is a DSC thermogram of Compound V Pattern 2 (Example 18). [Figure 88] 1 is a TGA thermogram of Compound V Pattern 2 (Example 18). [Figure 89] 1 is a comparison of XRPD diffractograms of Compound II Pattern 1 obtained from bulk stability studies (Example 19). [Figure 90] 1 is a comparison of XRPD diffractograms of Compound III Pattern 2 obtained from bulk stability studies (Example 19). [Figure 91] FIG. 1 shows the dynamic vapor sorption (DVS) plot and DVS change in mass plot of Compound II Pattern 1 (Example 21). [Figure 92] FIG. 1 shows a comparison of XRPD diffractograms of Compound II Pattern 1 before and after DVS studies (Example 21). [Figure 93] FIG. 1 shows the dynamic vapor sorption (DVS) plot and DVS change in mass plot of Compound III Pattern 2 (Example 21). [Figure 94] FIG. 1 shows a comparison of XRPD diffractograms of compound III pattern 2 before and after DVS studies (Example 21). [Figure 95] 1 is an XRPD diffractogram of Compound IV Pattern 1 obtained from Compound II Pattern 1 in Experiment PS4 of Example 22. [Figure 96] 1 is a DSC thermogram of Compound IV Pattern 1 obtained from Compound II Pattern 1 in Experiment PS4 of Example 22. [Figure 97] 1 is a TGA thermogram of Compound IV Pattern 1 obtained from Compound II Pattern 1 in Experiment PS4 of Example 22. [Figure 98] 1 is an XRPD diffractogram of Compound IV Pattern 2 obtained from Compound II Pattern 1 in Experiment PS5 of Example 22. [Figure 99] 1 is a DSC thermogram of Compound IV Pattern 2 obtained from Compound II Pattern 1 in Experiment PS5 of Example 22. [Figure 100] 1 is a TGA thermogram of Compound IV Pattern 2 obtained from Compound II Pattern 1 in Experiment PS5 of Example 22. [Figure 101]1 is an XRPD diffractogram of Compound III Pattern 3 obtained from Compound III Pattern 2 in Experiment PS3 of Example 22. [Figure 102] 1 is a DSC thermogram of Compound III Pattern 3 obtained from Compound III Pattern 2 in Experiment PS3 of Example 22. [Figure 103] 1 is a TGA thermogram of Compound III Pattern 3 obtained from Compound III Pattern 2 in Experiment PS3 of Example 22. [Figure 104] 1 is an XRPD diffractogram of Compound III Pattern 4 obtained from Compound III Pattern 2 in Experiment PS4 of Example 22. [Figure 105] 1 is a DSC thermogram of Compound III Pattern 4 obtained from Compound III Pattern 2 in Experiment PS4 of Example 22. [Figure 106] 1 is a TGA thermogram of Compound III Pattern 4 obtained from Compound III Pattern 2 in Experiment PS4 of Example 22. [Figure 107] FIG. 1 is an XRPD diffractogram of Compound III Pattern 5 obtained from Compound III Pattern 2 in Experiment PS5 of Example 22. [Figure 108] 1 is a DSC thermogram of Compound III Pattern 5 obtained from Compound III Pattern 2 in Experiment PS5 of Example 22. [Figure 109] 1 is a TGA thermogram of Compound III Pattern 5 obtained from Compound III Pattern 2 in Experiment PS5 of Example 22. [Figure 110] FIG. 1 is an XRPD diffractogram of Compound III Pattern 6 obtained from Compound III Pattern 2 in Experiment PS9 of Example 22. [Figure 111] 1 is a DSC thermogram of Compound III Pattern 6 obtained from Compound III Pattern 2 in Experiment PS9 of Example 22. [Figure 112] 1 is an XRPD diffractogram of Compound IV Pattern 1 obtained from Compound II Pattern 1 in Experiment PS8 of Example 22. [Figure 113]FIG. 1 shows a comparison of the XRPD diffraction patterns of Compound V Pattern 1 obtained from Compound III Pattern 2 in Experiment PS1 of Example 22 and Compound V Pattern 1 obtained in Example 18. [Figure 114] FIG. 2 shows the molecular structure of Compound II Pattern 1 determined by single crystal X-ray diffraction analysis in Example 25. [Figure 115] FIG. 1 shows the molecular structure of Compound II Pattern 1 as determined by single crystal X-ray diffraction analysis in Example 25. In the single crystalline form of Pattern 1, there is an intermolecular interaction between the protonated N5 atom of the free base and the O7 atom of the fumarate anion (N(5)-H(5)···O(7)). [Figure 116] Figure 1 shows an in vitro tissue penetration study comparing Compound II and ABI-1968 in vaginal tissue. Bar A shows the tissue penetration of 0.1% Compound II gel in porcine vaginal tissue. Bars B and C show the tissue penetration of 0.1% Compound II gel in human cervical tissue. Bar D shows the tissue penetration of a 1% formulation of ABI-1968 in 6% NMP into porcine vaginal tissue. Bar E shows the tissue penetration of a 1% nanosuspension of ABI-1968 in porcine vaginal tissue. Bar F shows the tissue penetration of a 3% formulation of ABI-1968 in 6% NMP into porcine vaginal tissue. Bar G shows the tissue penetration of a 3% formulation of ABI-1968 in 20% NMP into porcine vaginal tissue. ABI-1968 tissue penetration was significantly lower, hindering the compound's ability to reach HPV-infected cells. This may be a contributing factor to ABI-1968's performance in clinical studies. Surprisingly, Compound II exhibits high tissue penetration in both porcine and human tissues. High tissue penetration may translate into increased activity against HPV, as described in Example 41. [Figure 117] FIG. 1 is a process flow diagram for preparing a topical cream formulation as described in Example 29. [Figure 118] FIG. 1 is a flow diagram of the process for preparing the topical gel formulation described in Example 29. [Figure 119] FIG. 1 is a flow diagram of a process for preparing a tablet formulation as described in Example 30. [Figure 120]1 is an XRPD diffractogram of (S,S)-Compound I with moderate crystallinity as described in Example 12. [Figure 121] 1 is a bar graph comparing tissue penetration of the topical gel and topical tablet formulations described in Example 41. The tablet formulation results in similar tissue penetration as the topical gel, averaging 58 ng / mg of compound in the tissue. [Figure 122] 1 shows the structures of Compound I monofumarate, Compound II, and Compound III, the synthesis of which can be found in Examples 26-28. DETAILED DESCRIPTION OF THE INVENTION

[0066] It has been discovered that an effective composition for treating HPV infection or a disease or condition associated with HPV infection, such as HPV-induced neoplasia, including, but not limited to, cervical intraepithelial neoplasia, perianal intraepithelial neoplasia, penile intraepithelial neoplasia, vulvar intraepithelial neoplasia, anal intraepithelial neoplasia, and vaginal intraepithelial neoplasia, requires a selective combination of several aspects working in concert to achieve the desired result. To achieve the desired ability to penetrate epithelial layered tissue in effective amounts to deliver the active agent, it was essential to select an appropriate compound with favorable lipophilicity and tissue-penetrating properties, combined with a selected pharmaceutically acceptable salt (optionally in a favorable morphic form). After many failures, it took years of research to solve this problem for the benefit of patients worldwide who suffer from intraepithelial neoplasia, which has the potential to become cancerous.

[0067] Specifically, it has been discovered that the following particular salts are important compounds for delivering active agents: [ka]

[0068] Compound I is (ethyl((2-(2-amino-6-methoxy-9H-purin-9-yl)ethoxy)methyl)(benzyloxy)-phosphoryl)-L-alaninate. U.S. Patent No. 11,344,555, assigned to the Regents of the University of California, generally claims Compound I and pharmaceutically acceptable salts, and methods of using them in the treatment of papillomavirus infections. Compound I is an acyclic nucleotide phosphonate that is metabolized to a known, potent antiviral compound (PMEG; ((9-[2-phosphonomethoxy)ethyl)guanine]), but suffers from poor cell permeability and systemic toxicity that limits its use. The assignee has discovered an improved method for locally delivering prodrugs in a manner that is rapidly taken up by epithelial cells, a task that has been challenging to date and one that ABI-1968 failed to achieve.

[0069] Compound I (ethyl (-((2-(2-amino-6-methoxy-9H-purin-9-yl)ethoxy)methyl)-(benzyloxy)phosphoryl)-L-alaninate) has two asymmetric centers, one at the phosphorus atom and one at the amino acid moiety, each of which can be in the R or S configuration. Thus, Compound I has four stereoisomers. While U.S. Patent No. 5,629,599 and U.S. Patent No. 11,344,555 generally describe Compound I, these patents do not address the potential stereochemistry of the phosphorus atom. The stereoisomer of Compound I with R-stereochemistry at phosphorus and S-stereochemistry at the amino acid carbon has been discovered to have advantageous properties over the other three stereoisomers, as discussed further herein.

[0070] In a non-limiting embodiment, a preferred salt (e.g., fumarate) of Compound I is used as a mixture of (R,S) and (S,S) diastereomers, where the first R / S indicates the stereochemistry at the phosphorus atom and the second S indicates the stereochemistry of the carbon in the amino acid moiety (corresponding to an L-alanine residue with an S configuration). Any ratio of diastereomers that produces desirable results can be used, but the (R,S) diastereomer is prominent. In other embodiments, the ratio of R to S enantiomers at the phosphorus atom is approximately 1:1. In certain aspects, the compound is enantiomerically enriched in the R enantiomer at the phosphorus atom, with the amount of R by weight being, for example, greater than about 50%, or about 60% or more, 70% or more, 75% or more, 80% or more, or even 85% or more.

[0071] The S configuration of the asymmetric carbons, corresponding to the natural amino acid configuration, is advantageous in the present invention. In other aspects, the amount of S by weight is, for example, greater than about 50%, or greater than about 60%, 70%, 75%, 80%, or even 85%. In alternative embodiments, the R configuration of the chiral carbons predominates, greater than about 50%, or greater than about 60%, 70%, 75%, 80%, or even 85%.

[0072] Thus, in a primary aspect, the present invention provides an effective amount of the compounds described herein (R P , S C The present invention provides pharmaceutical salts of acyclic nucleotides, methods, compositions and dosage forms for the treatment of diseases associated with human papillomavirus (HPV). [ka]

[0073] The compounds, compositions, and dosage forms can also be used to treat conditions associated with or resulting from exposure to or infection with the HPV virus. For example, the active compounds can be used to treat precancerous cervical lesions, cervical intraepithelial neoplasia, vaginal, vulvar, penile, perianal, and anal intraepithelial neoplasia, cervical cancer, rectal cancer, penile cancer, vaginal cancer, and oropharyngeal cancer.

[0074] The active compounds and compositions can also be used to treat infections caused by a range of HPV types. Most cancer-causing HPV types are caused by alpha-7 and alpha-9 species, including types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 73, and 82. The most common cancer-causing HPV types are types 16 and 18. HPV-16 and HPV-18 are reported to be responsible for 50% of cervical cancers, and HPV-6 and HPV-11 are responsible for 90% of venereal warts (World Health Organization, "Cervical Cancer," https: / / www.who.int / news-room / fact-sheets / detail / cervical-cancer). Infection with one genotype does not preclude subsequent infection with a different genotype.

[0075] In one embodiment, Compound I monofumarate, Compound II, or Compound III is used to treat HPV-16. In one embodiment, Compound I monofumarate, Compound II, or Compound III is used to treat HPV-18. In one embodiment, Compound I monofumarate, Compound II, or Compound III is used to treat high-risk HPV infection. In one embodiment, Compound I monofumarate, Compound II, or Compound III is used to treat HPV types 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 73, or 82.

[0076] In one embodiment, the compound, formulation, or solid dosage form comprising the compound may also be used prophylactically to prevent or delay the progression of clinical disease in individuals who are HPV positive or have been exposed to HPV.

[0077] In particular, Compound II has been discovered to exhibit excellent drug-like and pharmacological properties.

[0078] Compound II has an R stereochemistry at the phosphorus atom, as confirmed by X-ray crystallography (see Example 25, Figures 114 and 115). In alternative embodiments, Compound II can be used in the form of any desired ratio of phosphorus R-enantiomer and S-enantiomer, up to and including pure enantiomers. In some embodiments, Compound II is used in a form that is at least 90% free of the opposite enantiomer, and may be at least 98%, 99%, or even 100% free of the opposite enantiomer. Unless otherwise specified, enantiomerically pure Compound II is at least 90% free of the opposite enantiomer. In certain embodiments, Compound II is used as a racemic mixture of isomers. Furthermore, in alternative embodiments, the amino acid of the phosphonamidate can be in the D or L configuration, or a mixture thereof, including a racemic mixture.

[0079] When the phosphonamidate exhibits chirality, it can be provided as an R- or S-chiral phosphorus derivative or mixtures thereof, including enantiomerically enriched forms, including racemic mixtures. All of these configurational combinations are alternative embodiments of the invention described herein. In another embodiment, at least one hydrogen atom of Compound I, Compound II, or Compound III can be replaced with deuterium.

[0080] In certain embodiments, compound I is [ka] or a pharmaceutically acceptable salt thereof.

[0081] In certain embodiments, compound II is [ka] It could be.

[0082] In certain embodiments, compound III is [ka] It could be.

[0083] I.(R P ,S C ) Ethyl (((2-(2-amino-6-methoxy-9H-purin-9-yl)ethoxy)methyl)(benzyloxy)phosphoryl)-L-alaninate fumarate (Compound II)

[0084] In certain embodiments, the active compound of the present invention is Compound II, which can be provided in its pharmaceutically acceptable composition or solid dosage form. In one embodiment, Compound II is an amorphous solid. In a further embodiment, Compound II is a crystalline solid.

[0085] Synthesis of Compound II The present invention further includes a non-limiting exemplary process for preparing a fumarate salt of Compound I, such as Compound II, comprising: (i) In a flask or container, R P , S C a first step of dissolving the isomer of formula (I) in an organic solvent, such as acetone, methanol, ethanol, isopropanol, dichloromethane, tetrahydrofuran or acetonitrile, (ii) R in step (i) P , S C adding fumaric acid to a solution of Compound I in a specific molar ratio (e.g., 0.5:1.0, 1.0:1.0, or 1.5:1) at ambient temperature or a slightly elevated or reduced temperature (e.g., 23°C to 55°C); (iii) stirring the reactants of step (ii) at ambient temperature or at a slightly elevated or reduced temperature; (iv) optionally seeding the solution of step (iii) with crystals of Compound II; (v) adding a second organic solvent, such as pentane, n-hexane, heptane, petroleum ether, methyl tert-butyl ether, diethyl ether or water to induce crystallization; (vi) optionally stirring the resulting solution at ambient temperature or at a slightly elevated or reduced temperature; (vii) cooling the obtained solution to a low temperature, for example, about 0°C to 10°C, and stirring the solution at that temperature; (viii) filtering the resulting solid; and (ix) optionally drying the solid under reduced pressure and elevated temperature, for example at or at least 30°C, 35°C, 40°C, 45°C or 50°C.

[0086] In certain embodiments, step (i) above is carried out in isopropanol. Additionally, the second organic solvent in step (v) may be, for example, heptane.

[0087] In one embodiment, R of compound I P , S C The isomer is dissolved in ethanol in step (i). In one embodiment, R P , S C The isomer is dissolved in methanol in step (i). In one embodiment, R P , S C In a further embodiment, the R isomer of compound I is dissolved in acetonitrile in step (i). P , S C The isomers are dissolved in tetrahydrofuran in step (i).

[0088] In one embodiment, the second organic solvent in step (v) is pentane. In one embodiment, the second organic solvent in step (v) is hexane. In one embodiment, the second organic solvent in step (v) is methyl tert-butyl ether. In one embodiment, the second organic solvent in step (v) is water.

[0089] The present invention further includes a non-limiting exemplary process for preparing compound III, which process comprises: (x) In a flask or container, S of Compound I P , S C a first step of dissolving the isomer of formula (I) in an organic solvent, such as acetone, methanol, ethanol, isopropanol, dichloromethane, tetrahydrofuran or acetonitrile, (xi) adding fumaric acid to the solution of Compound I from step (i) in a specific molar ratio (e.g., 0.5:1.0, 1.0:1.0, or 1.5:1) at ambient temperature or a slightly elevated or reduced temperature (e.g., about 23°C to 55°C); (xii) stirring the reactants of step (ii) at ambient temperature or at a slightly elevated or reduced temperature; (xiii) optionally seeding the solution of step (iii) with crystals of Compound II; (xiv) adding a second organic solvent, such as pentane, n-hexane, heptane, petroleum ether, methyl tert-butyl ether, diethyl ether or water to induce crystallization; (xv) optionally stirring the resulting solution at ambient temperature or at a slightly elevated or reduced temperature; (xvi) cooling the obtained solution to a low temperature, for example, about 0°C to 10°C, and stirring the solution at that temperature; (xvii) filtering the resulting solid; and (xviii) optionally drying the solid under reduced pressure and elevated temperature, for example at or at least 30°C, 35°C, 40°C, 45°C or 50°C.

[0090] In certain embodiments, step (i) above is carried out in isopropanol. Additionally, the second organic solvent in step (v) may be, for example, heptane.

[0091] In one embodiment, S of compound I P , S C In one embodiment, the S isomer of compound I is dissolved in ethanol in step (i). P , S C In one embodiment, the S isomer of compound I is dissolved in methanol in step (i). P , S C In a further embodiment, the S isomer of compound I is dissolved in acetonitrile in step (i). P , S C The isomers are dissolved in tetrahydrofuran in step (i).

[0092] In one embodiment, the second organic solvent in step (v) is pentane. In one embodiment, the second organic solvent in step (v) is hexane. In one embodiment, the second organic solvent in step (v) is methyl tert-butyl ether. In one embodiment, the second organic solvent in step (v) is water.

[0093] In certain embodiments, the monofumarate salt is synthesized from the sesquifumarate (1.5 equivalents of fumaric acid). The sesquifumarate can be washed with a solvent, such as methyl tertbutyl ether, to remove excess fumaric acid to provide the monofumarate.

[0094] II. Salts of Compound I In certain embodiments, the present invention provides compounds I, R P Compounds I and S P Compound I is provided as a monofumarate salt. In certain embodiments, the present invention provides Compound I, R P Compounds I and S P Compound I is provided as a hemifumarate salt. In certain embodiments, the present invention provides Compound I, R P Compounds I and SP Compound I is provided as a sesquifumarate salt. In certain embodiments, the present invention provides a compound of formula (I) P Compounds I and S P Compound I is provided as a sulfate salt. In certain embodiments, the present invention provides a compound of formula (I) P Compounds I and S P Compound I is provided as the hydrochloride salt. In certain embodiments, the present invention provides Compound I, R P Compounds I and S P Compound I is provided as a benzenesulfonate salt. In certain embodiments, the present invention provides a compound of formula (I) where R P Compounds I and S P Compound I is provided as a tosylate salt. In certain embodiments, the present invention provides a compound of formula I, R P Compounds I and S P Compound I is provided as the succinate salt. [ka] TIFF2025526212000013.tif210170

[0095] R of Compound I P salt [ka] TIFF2025526212000015.tif135170 Compound I S P salt [ka] TIFF2025526212000017.tif86170

[0096] The hemifumarate and monosuccinate salts form solids with advantageous properties for solid dosage forms for the treatment of hosts, such as humans, with HPV infections or HPV-associated diseases, such as intraepithelial neoplasias of the cervix, anus, penis, vulva, perianal, or vagina. However, the monofumarate exhibits superior properties to the hemifumarate and monosuccinate. Therefore, the monofumarate salt remains the preferred salt form of Compound I.

[0097] Additional embodiments of the present invention In certain embodiments, the present invention includes at least the following: 1.Formula: [ka] Compound.

[0098] 2.Formula: [ka] or a pharmaceutically acceptable salt thereof.

[0099] 3.Formula: [ka] or a pharmaceutically acceptable salt thereof.

[0100] 4.Formula: [ka] The compound of embodiment 2.

[0101] 5.Formula: [ka] The compound of embodiment 3.

[0102] 6. Formula of isolated morphic forms [ka] wherein the isolated morphic form is characterized by an XRPD pattern comprising peaks independently selected from at least three, four, five, or six of the following 2-theta values: 3.08±0.2°, 9.30±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 20.14±0.2°, 25.14±0.2°, and 28.82±0.2°.

[0103] 7. Formula of isolated morphic forms [ka] wherein the isolated morphic form is characterized by an XRPD pattern comprising peaks independently selected from at least three, four, five, six, seven, eight, or nine of the following 2-theta values: 9.53±0.2°, 10.04±0.2°, 11.60±0.2°, 14.57±0.2°, 17.22±0.2°, 17.50±0.2°, 20.04±0.2°, 20.36±0.2°, 22.34±0.2°, 23.73±0.2°, 25.48±0.2°, 26.06±0.2°, 27.38±0.2°, and 32.20±0.2°.

[0104] 8. Formula of isolated morphic forms [ka] wherein the isolated morphic form has the following 2-theta values: 8.94±0.2°, 9.89±0.2°, 9.91±0.2°, 11.66±0.2°, 12.11±0.2°, 15.13±0.2°, 17.85±0.2°, 18.15±0.2°, 19.90±0.2°, 20.38±0.2°, 22.94±0.2°, 25. A compound characterized by an XRPD pattern including peaks independently selected from at least three, four, five, six, seven, eight, or nine of: 26.09±0.2°, 26.54±0.2°, 26.90±0.2°, 27.38±0.2°, 28.28±0.2°, 28.95±0.2°, 29.64±0.2°, and 38.07±0.2°.

[0105] 9. Formula of isolated morphic form: [ka] wherein the isolated morphic form is characterized by an XRPD pattern comprising peaks independently selected from at least three, four, five, or six of the following 2-theta values: 3.08±0.2°, 9.30±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 20.14±0.2°, 25.14±0.2°, and 28.82±0.2°.

[0106] 10. A pharmaceutical composition comprising a compound according to any one of embodiments 1 to 5 and a pharmaceutically acceptable carrier.

[0107] 11. A pharmaceutical composition comprising the morphic form according to any one of embodiments 6 to 9 in a pharmaceutically acceptable carrier.

[0108] 12. The pharmaceutical composition of embodiment 10 or 11, which is in solid dosage form.

[0109] 13. The pharmaceutical composition according to embodiment 10 or 11, which is a semi-solid dosage form.

[0110] 14. The pharmaceutical composition according to embodiment 10 or 11, in the form of a reconstituted powder.

[0111] 15. The pharmaceutical composition according to embodiment 10 or 11, in the form of a dry powder dosage form.

[0112] 16. The pharmaceutical composition according to embodiment 10 or 11, which is in the form of a film.

[0113] 17. The pharmaceutical composition according to embodiment 10 or 11, which is in the form of a pessary.

[0114] 18. The pharmaceutical composition of embodiment 12, which is in the form of a tablet.

[0115] 19. The pharmaceutical composition according to embodiment 13, which is in the form of a cream.

[0116] 20. The pharmaceutical composition according to embodiment 13, which is in the form of a gel.

[0117] 21. The pharmaceutical composition according to any one of embodiments 10 to 20, formulated for topical administration.

[0118] 22. A pharmaceutical composition according to any one of embodiments 10 to 21, for delivery to the cervix.

[0119] 23. A pharmaceutical composition according to any one of embodiments 10 to 21, which is for vaginal delivery.

[0120] 24. A pharmaceutical composition according to any one of embodiments 10 to 21, for delivery to the vulva.

[0121] 25. A pharmaceutical composition according to any one of embodiments 10 to 21, for delivery to the perianal area.

[0122] 26. A pharmaceutical composition according to any one of embodiments 10 to 21, for delivery to the anus.

[0123] 27. A pharmaceutical composition according to any one of embodiments 10 to 21, for delivery to the penis.

[0124] 28. The pharmaceutical composition according to embodiment 18, wherein the tablet is a bilayer tablet.

[0125] 29. The pharmaceutical composition of embodiment 18, wherein the tablet disintegrates in less than about 250 μL of fluid.

[0126] 30. The pharmaceutical composition of embodiment 18, wherein the tablet disintegrates in less than about 150 μL of fluid.

[0127] 31. The pharmaceutical composition of any one of embodiments 10 to 30, comprising from about 0.001 mg to about 20 mg, from about 0.005 mg to about 10 mg, from about 0.01 mg to about 5 mg, from about 0.03 mg to about 1 mg, or from about 0.05 mg to about 0.3 mg of the compound.

[0128] 32. The pharmaceutical composition of any one of embodiments 10-30, comprising from about 0.005 mg to about 50 mg of the compound.

[0129] 33. The pharmaceutical composition of embodiment 32, comprising about 0.05 mg to about 40 mg of the compound.

[0130] 34. The pharmaceutical composition of embodiment 32, comprising about 0.1 mg to about 30 mg of the compound.

[0131] 35. The pharmaceutical composition of embodiment 32, comprising at least about 0.1 mg of the compound.

[0132] 36. The pharmaceutical composition of embodiment 32, comprising at least about 0.3 mg of the compound.

[0133] 37. The pharmaceutical composition of embodiment 32, comprising at least about 1 mg of the compound.

[0134] 38. The pharmaceutical composition of embodiment 32, comprising at least 1.5 mg of the compound.

[0135] 39. The pharmaceutical composition of embodiment 32, comprising at least about 2 mg of the compound.

[0136] 40. The pharmaceutical composition of any one of embodiments 19-29, comprising from about 0.001% to about 10% of the compound.

[0137] 41. The pharmaceutical composition of embodiment 40, comprising about 0.01% to 0.5% of the compound.

[0138] 42. The pharmaceutical composition of embodiment 40, comprising about 0.1% to 5% of the compound.

[0139] 43. A pharmaceutical composition according to any one of embodiments 10 to 42, comprising a mucoadhesive polymer.

[0140] 44. The pharmaceutical composition of embodiment 43, comprising about 5% to about 20% of a mucoadhesive polymer.

[0141] 45. The pharmaceutical composition of embodiment 43, comprising about 10% to about 50% mucoadhesive polymer.

[0142] 46. The pharmaceutical composition of embodiment 43, comprising about 50% to about 90% mucoadhesive polymer.

[0143] 47. The pharmaceutical composition of any one of embodiments 10-42, comprising a disintegration-enhancing excipient.

[0144] 48. A pharmaceutical composition according to any one of embodiments 10 to 42, comprising a penetration-enhancing excipient.

[0145] 49. A pharmaceutical composition according to any one of embodiments 10 to 42, comprising an excipient allowing controlled release of the active compound.

[0146] 50. The pharmaceutical composition of embodiment 19, wherein the pharmaceutically acceptable carrier is composed of light mineral oil, propylparaben, Tefose 63, water, EDTA, methylparaben, and Carbopol 974P.

[0147] 51. The pharmaceutical composition of embodiment 20, wherein the pharmaceutically acceptable carrier is comprised of water, EDTA, methylparaben, Carbopol 974P, propylene glycol, and sorbic acid.

[0148] 52. The pharmaceutical composition of embodiment 18, wherein the tablet is composed of mannitol, polycrystalline cellulose, and magnesium stearate.

[0149] 53. A method for treating a human papillomavirus infection, comprising administering to a host in need thereof an effective amount of a compound according to any one of embodiments 1-5, optionally in a pharmaceutically acceptable carrier.

[0150] 54. A method for treating a condition caused by human papillomavirus infection, comprising administering to a host in need thereof an effective amount of a compound according to any one of embodiments 1 to 5, optionally in a pharmaceutically acceptable carrier.

[0151] 55. The method of embodiment 54, wherein the condition caused by human papillomavirus infection is an intraepithelial neoplasia.

[0152] 56. The method of embodiment 55, wherein the condition caused by human papillomavirus is atypical squamous cells of undetermined significance (ASC-US).

[0153] 57. The method of embodiment 55, wherein the condition caused by human papillomavirus is atypical glandular cells (AGC).

[0154] 58. The method of embodiment 55, wherein the condition caused by human papillomavirus is low-grade squamous intraepithelial lesions (LSIL).

[0155] 59. The method of embodiment 55, wherein the condition caused by human papillomavirus is atypical squamous cells, high-grade squamous intraepithelial lesion cannot be excluded (ASC-H).

[0156] 60. The method of embodiment 55, wherein the condition caused by human papillomavirus is high grade squamous intraepithelial lesions (HSIL).

[0157] 61. The method of embodiment 54, wherein the condition caused by human papillomavirus is adenocarcinoma in situ (AIS).

[0158] 62. The method of embodiment 55, wherein the intraepithelial neoplasia is a cervical intraepithelial neoplasia.

[0159] 63. The method of embodiment 62, wherein the cervical intraepithelial neoplasia is grade 1 cervical intraepithelial neoplasia.

[0160] 64. The method of embodiment 62, wherein the cervical intraepithelial neoplasia is grade 2 cervical intraepithelial neoplasia.

[0161] 65. The method of embodiment 62, wherein the cervical intraepithelial neoplasia is grade 3 cervical intraepithelial neoplasia.

[0162] 66. The method of embodiment 55, wherein the intraepithelial neoplasia is a vaginal intraepithelial neoplasia.

[0163] 67. The method of embodiment 55, wherein the intraepithelial neoplasia is a vulvar intraepithelial neoplasia.

[0164] 68. The method of embodiment 55, wherein the intraepithelial neoplasia is an anal intraepithelial neoplasia.

[0165] 69. The method of embodiment 55, wherein the intraepithelial neoplasia is a perianal intraepithelial neoplasia.

[0166] 70. The method of embodiment 55, wherein the intraepithelial neoplasia is a penile intraepithelial neoplasia.

[0167] 71. The method of any one of embodiments 53 to 70, wherein the host is a human.

[0168] 72. The method of any one of embodiments 53-71, wherein the compound is administered topically.

[0169] 73. The method of any one of embodiments 52-71, wherein about 0.05 milligrams to about 40 milligrams of the compound is administered.

[0170] 74. The method of any one of embodiments 52-71, wherein about 0.1 milligrams to about 30 milligrams is administered.

[0171] 75. The method of any one of embodiments 52-71, wherein about 0.001 mg to about 20 mg, about 0.005 mg to about 10 mg, about 0.01 mg to about 5 mg, about 0.03 mg to about 1 mg, about 0.05 mg to about 0.3 mg, about 0.03 mg to about 0.07 mg, about 0.05 mg to about 0.15 mg, or about 0.15 mg to about 0.45 mg of the compound is administered.

[0172] 76. The method of embodiment 75, wherein about 0.05 mg to about 0.3 mg of the compound is administered.

[0173] 77. The method of any one of embodiments 52-76, further comprising applying a lubricating means to the epithelial tissue before inserting the dosage form into the affected area.

[0174] 78. The method of any one of embodiments 52-76, further comprising applying a lubricating means to the dosage form before inserting the dosage form into the affected area.

[0175] 79. The method of embodiment 77 or 78, wherein the lubricating means is selected from water, glycerol-based lubricants, and hydroxyethylcellulose-based lubricants.

[0176] 80. The method of any one of embodiments 52-79, wherein the compound is administered daily.

[0177] 81. The method of any one of embodiments 52-79, wherein the administration is twice daily.

[0178] 82. The method of any one of embodiments 52-79, wherein the administration is twice weekly.

[0179] 83. The method of any one of embodiments 52-79, administered three or more times per week.

[0180] 84. The method of any one of embodiments 52-83, wherein the administration is for about one week.

[0181] 85. The method of any one of embodiments 52-83, wherein the administration is for about 2 weeks.

[0182] 86. The method of any one of embodiments 52-83, wherein the administration is for about 3 weeks.

[0183] 87. The method of any one of embodiments 52-83, wherein the administration is for about 4 weeks.

[0184] 88. The method of any one of embodiments 52-83, wherein the administration is for about 5 weeks.

[0185] 89. The method of any one of embodiments 52-83, wherein the administration is for about 6 weeks.

[0186] 90. The compound is a. a treatment cycle comprising administration of the compound; and b. Drug-free cycles, including periods off treatment; 90. The method of any one of embodiments 52-89, wherein the treatment is administered in a treatment cycle comprising:

[0187] 91. The method of embodiment 90, wherein the drug holiday cycle is about 1 week.

[0188] 92. The method of embodiment 90, wherein the drug holiday cycle is about 2 weeks.

[0189] 93. The method of embodiment 90, wherein the drug holiday cycle is about 3 weeks.

[0190] 94. The method of any one of embodiments 52-79, wherein the compound is administered daily.

[0191] 95. The method of embodiment 94, wherein about 0.01 mg to about 0.5 mg is administered.

[0192] 96. The method of embodiment 95, wherein about 0.05 mg to about 0.3 mg is administered.

[0193] 97. The method of any one of embodiments 90-93, wherein two treatment cycles are applied.

[0194] 98. The method of any one of embodiments 90-93, wherein 3 treatment cycles are applied.

[0195] The method of any one of embodiments 90-93, wherein 99.4 treatment cycles are applied.

[0196] The method of any one of embodiments 90-93, wherein 100.5 treatment cycles are applied.

[0197] 101. The method of any one of embodiments 90-93, wherein 6 treatment cycles are applied.

[0198] 102. The method of any one of embodiments 52 to 101, wherein the human papillomavirus is HPV-16.

[0199] 103. The method of any one of embodiments 52 to 101, wherein the human papillomavirus is HPV-18.

[0200] 104. The method of any one of embodiments 52-103, wherein the compound is administered in combination with another antiviral compound.

[0201] 105. The method of embodiment 104, wherein the antiviral compound is selected from the group consisting of a protease inhibitor, another DNA polymerase inhibitor, an inhibitor of E6 or E6AP, an inhibitor of E7, an inhibitor of E1, an inhibitor of E2, an inhibitor of E1-E2 protein interaction, an L2 lipopeptide, an inhibitor of L1, an inhibitor of L2, a degrading agent of L1, and a degrading agent of L2.

[0202] 106. The method of any one of embodiments 52-103, wherein the compound is administered in combination with an anti-cancer compound.

[0203] 107. The method of embodiment 106, wherein the anti-cancer compound is selected from the group consisting of HDAC inhibitors, degraders of tetraspanins, immune checkpoint inhibitors, T-cell therapy, and anti-proliferative agents.

[0204] 108. The method of any one of embodiments 52-103, wherein the compound is administered in combination with a surgical procedure.

[0205] 109. The method of embodiment 108, wherein the compound is administered before a surgical procedure.

[0206] 110. The method of embodiment 108, wherein the compound is administered after a surgical procedure.

[0207] 111. The method of embodiment 108, wherein a surgical procedure is performed during administration of the compound.

[0208] 112. The method according to any one of embodiments 108 to 111, wherein the surgical procedure is resection of pathological tissue.

[0209] 113. The method of embodiment 112, wherein the excision is a loop electrosurgical excision procedure (LEEP).

[0210] 114. The method of embodiment 112, wherein the excision is a large loop excision of the transformation zone (LLETZ).

[0211] 115. The method of embodiment 112, wherein the excision is a knife conization.

[0212] 116. The method of embodiment 112, wherein the excision is laser conization.

[0213] 117. A method according to any one of embodiments 108 to 111, wherein the surgical procedure is cauterization of pathological tissue.

[0214] 118. The method of embodiment 117, wherein the ablation is laser ablation.

[0215] 119. The method of embodiment 117, wherein the cauterization is cryocauterization.

[0216] 120. Use of a compound according to embodiments 1-5, optionally in a pharmaceutically acceptable carrier, in the manufacture of a medicament for the treatment of a human papillomavirus infection in a host in need thereof.

[0217] 121. Use of a compound according to embodiments 1 to 5, optionally in a pharmaceutically acceptable carrier, in the manufacture of a medicament for the treatment of a condition caused by a human papillomavirus infection in a host in need thereof.

[0218] 122. The use according to embodiment 121, wherein the condition caused by human papillomavirus infection is an intraepithelial neoplasia.

[0219] 123. The use according to embodiment 122, wherein the intraepithelial neoplasia is a vaginal intraepithelial neoplasia.

[0220] 124. The use according to embodiment 122, wherein the intraepithelial neoplasia is a vulvar intraepithelial neoplasia.

[0221] 125. The use according to embodiment 122, wherein the intraepithelial neoplasia is a cervical intraepithelial neoplasia.

[0222] 126. The use according to embodiment 122, wherein the intraepithelial neoplasia is an anal intraepithelial neoplasia.

[0223] 127. The use according to embodiment 122, wherein the intraepithelial neoplasia is a perianal intraepithelial neoplasia.

[0224] 128. The use according to embodiment 122, wherein the intraepithelial neoplasia is a penile intraepithelial neoplasia.

[0225] 129. The use according to embodiments 120 to 128, wherein the host is a human.

[0226] 130. The use according to embodiments 120 to 129, which is for topical administration.

[0227] 131. A compound according to any one of embodiments 1-5, optionally in a pharmaceutically acceptable carrier, for use in the treatment of a human papillomavirus infection in a host in need thereof.

[0228] 132. A compound according to any one of embodiments 1 to 5, optionally in a pharmaceutically acceptable carrier, for use in the treatment of a condition caused by a human papillomavirus infection in a host in need thereof.

[0229] 133. The compound for use according to embodiment 132, wherein the condition caused by human papillomavirus infection is an intraepithelial neoplasia.

[0230] 134. The compound for use according to embodiment 133, wherein the intraepithelial neoplasia is a vaginal intraepithelial neoplasia.

[0231] 135. The compound for use according to embodiment 133, wherein the intraepithelial neoplasia is vulvar intraepithelial neoplasia.

[0232] 136. The compound for use according to embodiment 133, wherein the intraepithelial neoplasia is a cervical intraepithelial neoplasia.

[0233] 137. The compound for use according to embodiment 133, wherein the intraepithelial neoplasia is anal intraepithelial neoplasia.

[0234] 138. The compound for use according to embodiment 133, wherein the intraepithelial neoplasia is a perianal intraepithelial neoplasia.

[0235] 139. The compound for use according to embodiment 133, wherein the intraepithelial neoplasia is a penile intraepithelial neoplasia.

[0236] 140. The compound for use according to embodiments 131 to 139, wherein the host is a human.

[0237] 141. The compound for use according to embodiments 131-140, wherein the compound is administered topically.

[0238] 142. aR P Dissolving Compound I in an alcohol solvent; b. stirring at a temperature of about 20°C to about 70°C; c. adding 1.0 equivalent of fumaric acid; d. adding an aliphatic solvent; e. cooling the mixture; f. stirring the cooled solution; g. isolating and drying the solid; 5. A method for preparing the morphic form of embodiment 4, comprising:

[0239] 143. The method of embodiment 142, wherein the alcohol solvent in step (a) is ethanol or isopropanol.

[0240] 144. The method of embodiment 142 or 143, wherein the alcohol solvent in step (a) is isopropanol.

[0241] 145. The method of embodiment 142, wherein the solution of step (b) is stirred at about 45°C to about 55°C.

[0242] 146. The method of embodiment 142, wherein the aliphatic solvent is hexane or heptane.

[0243] 147. The method of embodiments 142 and 146, wherein the aliphatic solvent is heptane.

[0244] 148. The method of embodiment 142, wherein the mixture is cooled to below about 20°C.

[0245] 149. The method of embodiment 142, wherein the mixture is cooled to less than about 10°C.

[0246] 150. The method of embodiment 142, wherein the mixture is cooled to less than about 5°C.

[0247] 151. The method of embodiment 142, wherein the mixture is cooled to about 5°C to 0°C.

[0248] III. Morphic Form The present invention provides an isolated morphic form of Compound I monofumarate, Pattern 1. In certain embodiments, Compound I monofumarate Pattern 1 is characterized by the XRPD pattern in Figure 23, or a pattern substantially similar thereto.

[0249] 1. In one embodiment, Compound I monofumarate pattern 1 is characterized by an XRPD pattern comprising at least about three, four, or five 2-theta values selected from 6.0±0.2°, 8.9±0.2°, 9.3±0.2°, 9.7±0.2°, 11.9±0.2°, 14.8±0.2°, 18.0±0.2°, 20.0±0.2°, 23.4±0.2°, 25.2±0.2°, 25.9±0.2°, 26.8±0.2°, and 28.0±0.2°.

[0250] 2. In one embodiment, Compound I monofumarate pattern 1 is characterized by an XRPD pattern comprising at least 12 2-theta values selected from 6.0±0.2°, 8.9±0.2°, 9.3±0.2°, 9.7±0.2°, 11.9±0.2°, 14.8±0.2°, 18.0±0.2°, 20.0±0.2°, 23.4±0.2°, 25.2±0.2°, 25.9±0.2°, 26.8±0.2°, and 28.0±0.2°.

[0251] 3. In one embodiment, Compound I monofumarate pattern 1 is characterized by an XRPD pattern comprising at least 11 2-theta values selected from 6.0±0.2°, 8.9±0.2°, 9.3±0.2°, 9.7±0.2°, 11.9±0.2°, 14.8±0.2°, 18.0±0.2°, 20.0±0.2°, 23.4±0.2°, 25.2±0.2°, 25.9±0.2°, 26.8±0.2°, and 28.0±0.2°.

[0252] 4. In one embodiment, Compound I monofumarate pattern 1 is characterized by an XRPD pattern comprising at least ten 2-theta values selected from 6.0±0.2°, 8.9±0.2°, 9.3±0.2°, 9.7±0.2°, 11.9±0.2°, 14.8±0.2°, 18.0±0.2°, 20.0±0.2°, 23.4±0.2°, 25.2±0.2°, 25.9±0.2°, 26.8±0.2°, and 28.0±0.2°.

[0253] 5. In one embodiment, Compound I monofumarate pattern 1 is characterized by an XRPD pattern comprising at least nine 2-theta values selected from 6.0±0.2°, 8.9±0.2°, 9.3±0.2°, 9.7±0.2°, 11.9±0.2°, 14.8±0.2°, 18.0±0.2°, 20.0±0.2°, 23.4±0.2°, 25.2±0.2°, 25.9±0.2°, 26.8±0.2°, and 28.0±0.2°.

[0254] 6. In one embodiment, Compound I monofumarate pattern 1 is characterized by an XRPD pattern comprising at least eight 2-theta values selected from 6.0±0.2°, 8.9±0.2°, 9.3±0.2°, 9.7±0.2°, 11.9±0.2°, 14.8±0.2°, 18.0±0.2°, 20.0±0.2°, 23.4±0.2°, 25.2±0.2°, 25.9±0.2°, 26.8±0.2°, and 28.0±0.2°.

[0255] 7. In one embodiment, Compound I monofumarate pattern 1 is characterized by an XRPD pattern comprising at least seven 2-theta values selected from 6.0±0.2°, 8.9±0.2°, 9.3±0.2°, 9.7±0.2°, 11.9±0.2°, 14.8±0.2°, 18.0±0.2°, 20.0±0.2°, 23.4±0.2°, 25.2±0.2°, 25.9±0.2°, 26.8±0.2°, and 28.0±0.2°.

[0256] 8. In one embodiment, Compound I monofumarate pattern 1 is characterized by an XRPD pattern comprising at least six 2-theta values selected from 6.0±0.2°, 8.9±0.2°, 9.3±0.2°, 9.7±0.2°, 11.9±0.2°, 14.8±0.2°, 18.0±0.2°, 20.0±0.2°, 23.4±0.2°, 25.2±0.2°, 25.9±0.2°, 26.8±0.2°, and 28.0±0.2°.

[0257] 9. In one embodiment, Compound I monofumarate pattern 1 is characterized by an XRPD pattern comprising at least five 2-theta values selected from 6.0±0.2°, 8.9±0.2°, 9.3±0.2°, 9.7±0.2°, 11.9±0.2°, 14.8±0.2°, 18.0±0.2°, 20.0±0.2°, 23.4±0.2°, 25.2±0.2°, 25.9±0.2°, 26.8±0.2°, and 28.0±0.2°.

[0258] 10. In one embodiment, Compound I monofumarate pattern 1 is characterized by an XRPD pattern comprising at least four 2-theta values selected from 6.0±0.2°, 8.9±0.2°, 9.3±0.2°, 9.7±0.2°, 11.9±0.2°, 14.8±0.2°, 18.0±0.2°, 20.0±0.2°, 23.4±0.2°, 25.2±0.2°, 25.9±0.2°, 26.8±0.2°, and 28.0±0.2°.

[0259] 11. In one embodiment, Compound I monofumarate pattern 1 is characterized by an XRPD pattern comprising at least three 2-theta values selected from 6.0±0.2°, 8.9±0.2°, 9.3±0.2°, 9.7±0.2°, 11.9±0.2°, 14.8±0.2°, 18.0±0.2°, 20.0±0.2°, 23.4±0.2°, 25.2±0.2°, 25.9±0.2°, 26.8±0.2°, and 28.0±0.2°.

[0260] Compound I monofumarate Pattern 1 can be prepared, for example, by crystallization from isopropyl alcohol and heptane, as described in Example 7. Compound I free base and about 1.0 equivalent of fumaric acid can be dissolved in isopropanol at a concentration of, for example, about 25% to about 40% weight / volume and stirred at an elevated temperature, for example, about 45°C, about 50°C, or about 55°C. The solution is stirred at this temperature until some solids form, and then optionally seeded with Compound I monofumarate Pattern 1 crystalline solid. The mixture is stirred and cooled to a lower temperature, for example, below about 40°C, below about 30°C, below about 25°C, below about 20°C, or below about 15°C, to promote crystallization. The mixture is then stirred while heptane is added in an amount ranging from about 1 mL per mL of isopropanol to about 5 mL per mL of isopropanol (e.g., about 4 mL per mL of isopropanol). The resulting suspension is stirred, for example, at 25°C for at least about 24 hours while the product crystallizes. The suspension is then cooled to further promote crystallization. The solution can be cooled to less than about 10°C, less than about 5°C, less than about 0°C, or less than about -5°C. After stirring the solution at the lower temperature and allowing time for additional product to crystallize, for example, at least about 1 day, the solid is collected by filtration. The collected solid is dried under reduced pressure, and optionally, dried at elevated temperature, to provide Compound I monofumarate Pattern 1.

[0261] The present invention provides an isolated morphic form of Compound II, Pattern 1. In one embodiment, Pattern 1 is characterized by the XRPD pattern in Figure 71, or a pattern substantially similar thereto.

[0262] 1. In one embodiment, Compound II Pattern 1 is characterized by an XRPD pattern comprising at least three, four, or five 2-theta values selected from 3.08±0.2°, 9.30±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 20.14±0.2°, 25.14±0.2°, and 28.82±0.2°.

[0263] 2. Compound II pattern 1 of embodiment 1, characterized by an XRPD pattern comprising at least seven 2-theta values selected from 3.08±0.2°, 9.30±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 20.14±0.2°, 25.14±0.2°, and 28.82±0.2°.

[0264] 3. Compound II Pattern 1 of embodiment 1, characterized by an XRPD pattern comprising at least six 2-theta values selected from 3.08±0.2°, 9.30±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 20.14±0.2°, 25.14±0.2°, and 28.82±0.2°.

[0265] 4. Compound II Pattern 1 of embodiment 1, characterized by an XRPD pattern comprising at least five 2-theta values selected from: 3.08±0.2°, 9.30±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 20.14±0.2°, 25.14±0.2°, and 28.82±0.2°.

[0266] 5. Compound II Pattern 1 of embodiment 1, characterized by an XRPD pattern comprising at least four 2-theta values selected from: 5.3.08±0.2°, 9.30±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 20.14±0.2°, 25.14±0.2°, and 28.82±0.2°.

[0267] 6. Compound II Pattern 1 of embodiment 1, characterized by an XRPD pattern comprising at least three 2-theta values selected from: 6.3.08±0.2°, 9.30±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 20.14±0.2°, 25.14±0.2°, and 28.82±0.2°.

[0268] 2. Compound II pattern 1 of embodiment 1, characterized by an XRPD pattern comprising at least two 2-theta values selected from 7.3.08±0.2°, 9.30±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 20.14±0.2°, 25.14±0.2°, and 28.82±0.2°.

[0269] 2. Compound II pattern 1 of embodiment 1, characterized by an XRPD pattern comprising at least one 2-theta value selected from 8.3.08±0.2°, 9.30±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 20.14±0.2°, 25.14±0.2°, and 28.82±0.2°.

[0270] 9. In one embodiment, Compound II Pattern 1 is characterized by an XRPD pattern comprising at least three, four, or five 2-theta values selected from 3.08±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 18.13±0.2°, 19.78±0.2°, 20.14±0.2°, 22.91±0.2°, 23.34±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27.99±0.2°, and 28.82±0.2°.

[0271] 10. Compound II pattern 1 of embodiment 9, characterized by an XRPD pattern comprising at least 15 2-theta values selected from 10.3.08±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 18.13±0.2°, 19.78±0.2°, 20.14±0.2°, 22.91±0.2°, 23.34±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27.99±0.2°, and 28.82±0.2°.

[0272] 11. Compound II pattern 1 of embodiment 9, characterized by an XRPD pattern comprising at least 14 2-theta values selected from: 11.3.08±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 18.13±0.2°, 19.78±0.2°, 20.14±0.2°, 22.91±0.2°, 23.34±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27.99±0.2°, and 28.82±0.2°.

[0273] 10. Compound II pattern 1 of embodiment 9, characterized by an XRPD pattern comprising at least 13 2-theta values selected from: 12.3.08±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 18.13±0.2°, 19.78±0.2°, 20.14±0.2°, 22.91±0.2°, 23.34±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27.99±0.2°, and 28.82±0.2°.

[0274] 10. Compound II pattern 1 of embodiment 9, characterized by an XRPD pattern comprising at least 12 2-theta values selected from 13.3.08±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 18.13±0.2°, 19.78±0.2°, 20.14±0.2°, 22.91±0.2°, 23.34±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27.99±0.2°, and 28.82±0.2°.

[0275] 10. Compound II pattern 1 of embodiment 9, characterized by an XRPD pattern comprising at least 11 2-theta values selected from 14.3.08±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 18.13±0.2°, 19.78±0.2°, 20.14±0.2°, 22.91±0.2°, 23.34±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27.99±0.2°, and 28.82±0.2°.

[0276] 10. Compound II pattern 1 of embodiment 9, characterized by an XRPD pattern comprising at least ten 2-theta values selected from 15.3.08±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 18.13±0.2°, 19.78±0.2°, 20.14±0.2°, 22.91±0.2°, 23.34±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27.99±0.2°, and 28.82±0.2°.

[0277] 10. Compound II pattern 1 of embodiment 9, characterized by an XRPD pattern comprising at least nine 2-theta values selected from 16.3.08±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 18.13±0.2°, 19.78±0.2°, 20.14±0.2°, 22.91±0.2°, 23.34±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27.99±0.2°, and 28.82±0.2°.

[0278] 10. Compound II pattern 1 of embodiment 9, characterized by an XRPD pattern comprising at least eight 2-theta values selected from 17.3.08±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 18.13±0.2°, 19.78±0.2°, 20.14±0.2°, 22.91±0.2°, 23.34±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27.99±0.2°, and 28.82±0.2°.

[0279] 18. In certain embodiments, Compound II Pattern 1 has the following peaks: 3.08±0.2°, 6.07±0.2°, 8.80±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 12.60±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 17.838±0.2°, 18.13± 0.2°, 18.63±0.2°, 18.89±0.2°, 19.78±0.2°, 20.14±0.2°, 20.81±0.2°, 21.24±0.2°, 21.59±0.2°, 21.89±0.2°, 22.91±0.2°, 23.34±0.2°, 24.23±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86 ±0.2°, 26.78±0.2°, 27.13±0.2°, 27.59±0.2°, 27.99±0.2°, 28.82±0.2°, 29.23±0.2°, 29.45±0.2°, 30.39±0.2°, 31.18±0.2°, 31.66±0.2°, 32.27±0.2°, 32.69±0.2°, 33.43±0.2°, 34.0 and 39.24±0.2°.

[0280] 19.3.08±0.2°, 6.07±0.2°, 8.80±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 12.60±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 17.838±0.2°, 18.13±0.2°, 18.63±0.2°, 18.89±0.2° .2°, 19.78±0.2°, 20.14±0.2°, 20.81±0.2°, 21.24±0.2°, 21.59±0.2°, 21.89±0.2°, 22.91±0.2°, 23.34±0.2°, 24.23±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27. 13±0.2°, 27.59±0.2°, 27.99±0.2°, 28.82±0.2°, 29.23±0.2°, 29.45±0.2°, 30.39±0.2°, 31.18±0.2°, 31.66±0.2°, 32.27±0.2°, 32.69±0.2°, 33.43±0.2°, 34.06±0.2°, 34.34±0.2° 19. Compound II pattern 1 of embodiment 18, characterized by an XRPD pattern comprising at least 45 2-theta values selected from: 34.69±0.2°, 35.58±0.2°, 36.19±0.2°, 36.62±0.2°, 37.41±0.2°, 38.30±0.2°, 38.77±0.2°, and 39.24±0.2°.

[0281] 20.3.08±0.2°, 6.07±0.2°, 8.80±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 12.60±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 17.838±0.2°, 18.13±0.2°, 18.63±0.2°, 18.89±0.2° .2°, 19.78±0.2°, 20.14±0.2°, 20.81±0.2°, 21.24±0.2°, 21.59±0.2°, 21.89±0.2°, 22.91±0.2°, 23.34±0.2°, 24.23±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27. 13±0.2°, 27.59±0.2°, 27.99±0.2°, 28.82±0.2°, 29.23±0.2°, 29.45±0.2°, 30.39±0.2°, 31.18±0.2°, 31.66±0.2°, 32.27±0.2°, 32.69±0.2°, 33.43±0.2°, 34.06±0.2°, 34.34±0.2° 19. Compound II pattern 1 of embodiment 18, characterized by an XRPD pattern comprising at least 40 2-theta values selected from: 34.69±0.2°, 35.58±0.2°, 36.19±0.2°, 36.62±0.2°, 37.41±0.2°, 38.30±0.2°, 38.77±0.2°, and 39.24±0.2°.

[0282] 21.3.08±0.2°, 6.07±0.2°, 8.80±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 12.60±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 17.838±0.2°, 18.13±0.2°, 18.63±0.2°, 18.89±0.2° .2°, 19.78±0.2°, 20.14±0.2°, 20.81±0.2°, 21.24±0.2°, 21.59±0.2°, 21.89±0.2°, 22.91±0.2°, 23.34±0.2°, 24.23±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27. 13±0.2°, 27.59±0.2°, 27.99±0.2°, 28.82±0.2°, 29.23±0.2°, 29.45±0.2°, 30.39±0.2°, 31.18±0.2°, 31.66±0.2°, 32.27±0.2°, 32.69±0.2°, 33.43±0.2°, 34.06±0.2°, 34.34±0.2° 19. Compound II pattern 1 of embodiment 18, characterized by an XRPD pattern comprising at least 35 2-theta values selected from: 34.69±0.2°, 35.58±0.2°, 36.19±0.2°, 36.62±0.2°, 37.41±0.2°, 38.30±0.2°, 38.77±0.2°, and 39.24±0.2°.

[0283] 22.3.08±0.2°, 6.07±0.2°, 8.80±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 12.60±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 17.838±0.2°, 18.13±0.2°, 18.63±0.2°, 18.89±0.2° .2°, 19.78±0.2°, 20.14±0.2°, 20.81±0.2°, 21.24±0.2°, 21.59±0.2°, 21.89±0.2°, 22.91±0.2°, 23.34±0.2°, 24.23±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27. 13±0.2°, 27.59±0.2°, 27.99±0.2°, 28.82±0.2°, 29.23±0.2°, 29.45±0.2°, 30.39±0.2°, 31.18±0.2°, 31.66±0.2°, 32.27±0.2°, 32.69±0.2°, 33.43±0.2°, 34.06±0.2°, 34.34±0.2° 19. Compound II pattern 1 of embodiment 18, characterized by an XRPD pattern comprising at least 30 2-theta values selected from: 34.69±0.2°, 35.58±0.2°, 36.19±0.2°, 36.62±0.2°, 37.41±0.2°, 38.30±0.2°, 38.77±0.2°, and 39.24±0.2°.

[0284] 23.3.08±0.2°, 6.07±0.2°, 8.80±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 12.60±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 17.838±0.2°, 18.13±0.2°, 18.63±0.2°, 18.89±0.2° .2°, 19.78±0.2°, 20.14±0.2°, 20.81±0.2°, 21.24±0.2°, 21.59±0.2°, 21.89±0.2°, 22.91±0.2°, 23.34±0.2°, 24.23±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27. 13±0.2°, 27.59±0.2°, 27.99±0.2°, 28.82±0.2°, 29.23±0.2°, 29.45±0.2°, 30.39±0.2°, 31.18±0.2°, 31.66±0.2°, 32.27±0.2°, 32.69±0.2°, 33.43±0.2°, 34.06±0.2°, 34.34±0.2° 19. Compound II pattern 1 of embodiment 18, characterized by an XRPD pattern comprising at least 25 2-theta values selected from: 34.69±0.2°, 35.58±0.2°, 36.19±0.2°, 36.62±0.2°, 37.41±0.2°, 38.30±0.2°, 38.77±0.2°, and 39.24±0.2°.

[0285] 24.3.08±0.2°, 6.07±0.2°, 8.80±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 12.60±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 17.838±0.2°, 18.13±0.2°, 18.63±0.2°, 18.89±0.2° .2°, 19.78±0.2°, 20.14±0.2°, 20.81±0.2°, 21.24±0.2°, 21.59±0.2°, 21.89±0.2°, 22.91±0.2°, 23.34±0.2°, 24.23±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27. 13±0.2°, 27.59±0.2°, 27.99±0.2°, 28.82±0.2°, 29.23±0.2°, 29.45±0.2°, 30.39±0.2°, 31.18±0.2°, 31.66±0.2°, 32.27±0.2°, 32.69±0.2°, 33.43±0.2°, 34.06±0.2°, 34.34±0.2° 19. Compound II pattern 1 of embodiment 18, characterized by an XRPD pattern comprising at least 20 2-theta values selected from: 34.69±0.2°, 35.58±0.2°, 36.19±0.2°, 36.62±0.2°, 37.41±0.2°, 38.30±0.2°, 38.77±0.2°, and 39.24±0.2°.

[0286] 25.3.08±0.2°, 6.07±0.2°, 8.80±0.2°, 9.30±0.2°, 10.66±0.2°, 12.08±0.2°, 12.60±0.2°, 14.92±0.2°, 15.10±0.2°, 17.45±0.2°, 17.838±0.2°, 18.13±0.2°, 18.63±0.2°, 18.89±0.2° .2°, 19.78±0.2°, 20.14±0.2°, 20.81±0.2°, 21.24±0.2°, 21.59±0.2°, 21.89±0.2°, 22.91±0.2°, 23.34±0.2°, 24.23±0.2°, 25.14±0.2°, 25.33±0.2°, 25.86±0.2°, 26.78±0.2°, 27. 13±0.2°, 27.59±0.2°, 27.99±0.2°, 28.82±0.2°, 29.23±0.2°, 29.45±0.2°, 30.39±0.2°, 31.18±0.2°, 31.66±0.2°, 32.27±0.2°, 32.69±0.2°, 33.43±0.2°, 34.06±0.2°, 34.34±0.2° 19. Compound II pattern 1 of embodiment 18, characterized by an XRPD pattern comprising at least 15 2-theta values selected from: 34.69±0.2°, 35.58±0.2°, 36.19±0.2°, 36.62±0.2°, 37.41±0.2°, 38.30±0.2°, 38.77±0.2°, and 39.24±0.2°.

[0287] 26. What is the ratio of compound II to fumaric acid? 1 26. Compound II Pattern 1 according to any one of embodiments 1 to 25, which is approximately 1:1 by H NMR.

[0288] Compound II Pattern 1 can be prepared, for example, by recrystallizing Compound II (Example 12, Table 31), by equilibrating Compound II in a suitable solvent, or by crystallization by slow evaporation of the solvent (Example 12, Table 32). Thus, the present invention includes at least the following features: 1. In certain embodiments, Compound II Pattern 1 is prepared by recrystallization of Compound II. 2. In certain embodiments, compound II pattern 1 is prepared by equilibration of compound II in a solvent. 3. In certain embodiments, Compound II Pattern 1 is prepared by slow evaporation of solvent from a solution of Compound II. 4. The process of embodiment 1, wherein compound II is dissolved in an alcohol solvent and crystallized as pattern 1 by addition of an ether solvent. 5. The process of embodiment 1, wherein compound II is dissolved in an alcohol solvent and crystallized as pattern 1 by addition of an aliphatic solvent. 6. The process of embodiment 1, wherein compound II is dissolved in an ether solvent and crystallized as pattern 1 by addition of an aliphatic solvent. 7. The process of embodiment 4 or 5, wherein the alcoholic solvent is or comprises methanol. 8. The process of embodiment 4 or 5, wherein the alcoholic solvent is or comprises ethanol. 9. The process of embodiment 4 or 5, wherein the alcohol solvent is or comprises isopropanol. 10. The process of embodiment 4 or 5, wherein the alcohol solvent is or comprises n-propanol. 11. The process of embodiment 4 or 5, wherein the alcohol solvent is or comprises n-butanol. 12. The process of embodiment 4 or 5, wherein the alcohol solvent is or comprises isoamyl alcohol. 13. The process of embodiment 4 or 5, wherein the alcohol solvent is or comprises cyclohexanol. 14. The process of embodiment 4 or 6, wherein the ether solvent is or comprises diethyl ether. 15. The process of embodiment 4 or 6, wherein the ether solvent is or comprises dibutyl ether. 16. The process of embodiment 4 or 6, wherein the ether solvent is or comprises methyl tert-butyl ether. 17. The process of embodiment 4 or 6, wherein the ether solvent is or comprises cyclopropyl methyl ether. 18. The process of embodiment 4 or 6, wherein the ether solvent is or comprises a glyme. 19. The process of embodiment 4 or 6, wherein the ether solvent is tetrahydrofuran or the ether solvent comprises tetrahydrofuran. 20. The process of embodiment 4 or 6, wherein the ether solvent is or comprises 2-methyltetrahydrofuran. 21. The process of embodiment 4 or 6, wherein the ether solvent is or comprises dioxane. 22. The process of any one of embodiments 5-21, wherein the aliphatic solvent is pentane or comprises pentane. 23. The process of any one of embodiments 5-21, wherein the aliphatic solvent is or comprises n-hexane. 24. The process of any one of embodiments 5-21, wherein the aliphatic solvent is heptane or comprises heptane. 25. The process of any one of embodiments 5-21, wherein the aliphatic solvent is or comprises petroleum ether. 26. The process of any one of embodiments 5-21, wherein the aliphatic solvent is octane or comprises octane. 27. The process of any one of embodiments 5-21, wherein the aliphatic solvent is cyclohexane or comprises cyclohexane. 28. The process of any one of embodiments 5-21, wherein the aliphatic solvent is or comprises a mixture of hexane isomers. 29. The process of any one of embodiments 4 to 28, wherein the solids are collected by filtration. 30. The process of any one of embodiments 4 to 28, wherein after adding the aliphatic solvent, the solution is cooled. 31. The process of embodiment 30, wherein the solution is cooled to less than about 10°C. 32. The process of embodiment 30, wherein the solution is cooled to about 5°C or below 5°C. 33. The process of any one of embodiments 4 to 32, wherein the solution, suspension, or slurry is stirred until crystallization occurs. 34. The process of embodiment 33, wherein the solution, suspension, or slurry is stirred for at least about 1 day. 35. The process of embodiment 33, wherein the solution, suspension, or slurry is stirred for at least about 1 week. 36. The process of embodiment 33, wherein the solution, suspension, or slurry is stirred for at least about 2 weeks. 37. The process of embodiment 33, wherein the solution, suspension, or slurry is stirred for at least about 3 weeks. 38. The process of embodiment 2, wherein compound II is dissolved in the equilibration solvent at 25°C and stirred for a period of about 2 weeks to about 3 weeks. 39. The process of embodiment 2, wherein the equilibration solvent is a binary mixture of solvents. 40. The process of embodiment 2, wherein the equilibration solvent is isopropanol. 41. The process of embodiment 39, wherein the equilibration solvent is about 1 part tetrahydrofuran to about 3 parts heptane. 42. The process of embodiment 39, wherein the equilibration solvent is about 1 part ethanol to about 3 parts heptane. 43. The process of embodiment 39, wherein the equilibration solvent is about 1 part ethyl acetate to about 3 parts toluene. 44. The process of embodiment 39, wherein the equilibration solvent is about 1 part isopropanol to about 1 part heptane. 45. The process of embodiment 39, wherein the equilibration solvent is about 1 part isopropanol to about 1 part toluene. 46. The process of embodiment 39, wherein the equilibration solvent is about 3 parts methyl tert-butyl ether to about 1 part isopropanol. 47. The process of embodiment 39, wherein the equilibration solvent is about 1 part isopropanol to about 4 parts heptane. 48. The process of embodiment 39, wherein the equilibration solvent is about 1 part ethanol to about 1 part toluene. 49. The process of embodiment 3, wherein compound II is dissolved in a suitable solvent, filtered through a 0.45 μM filter, and then maintained at 23° C. and 1 atmosphere until the solvent evaporates. 50. The process of embodiment 49, wherein the solvent is or comprises acetone. 51. The process of embodiment 49, wherein the solvent is or comprises methyl ethyl ketone. 52. The process of embodiment 49, wherein the solvent is or comprises ethyl acetate. 53. The process of embodiment 49, wherein the solvent is or comprises methanol. 54. The process of embodiment 49, wherein the solvent is or comprises ethanol. 55. The process of embodiment 49, wherein the solvent is or comprises isopropanol. 56. The process of embodiment 49, wherein the solvent is or comprises tetrahydrofuran.

[0289] Provided herein are isolated morphic forms of Compound III, Pattern 1 and Pattern 2. In one embodiment, Pattern 1 is characterized by the XRPD pattern in Figure 77, or a pattern substantially similar thereto. 1. In one embodiment, Compound III Pattern 1 is characterized by an XRPD pattern comprising at least two, three, four, five, or six 2-theta values independently selected from 9.53±0.2°, 10.04±0.2°, 11.60±0.2°, 14.57±0.2°, 17.22±0.2°, 17.50±0.2°, 20.04±0.2°, 20.36±0.2°, 22.34±0.2°, 23.73±0.2°, 25.48±0.2°, 26.06±0.2°, 27.38±0.2°, and 32.20±0.2°. 2. Pattern 1 of compound III of embodiment 1, characterized by an XRPD pattern comprising at least 12 2-theta values selected from 9.53±0.2°, 10.04±0.2°, 11.60±0.2°, 14.57±0.2°, 17.22±0.2°, 17.50±0.2°, 20.04±0.2°, 20.36±0.2°, 22.34±0.2°, 23.73±0.2°, 25.48±0.2°, 26.06±0.2°, 27.38±0.2°, and 32.20±0.2°. 3. Pattern 1 of compound III of embodiment 1, characterized by an XRPD pattern comprising at least 11 2-theta values selected from: 9.53±0.2°, 10.04±0.2°, 11.60±0.2°, 14.57±0.2°, 17.22±0.2°, 17.50±0.2°, 20.04±0.2°, 20.36±0.2°, 22.34±0.2°, 23.73±0.2°, 25.48±0.2°, 26.06±0.2°, 27.38±0.2°, and 32.20±0.2°. 4. Pattern 1 of compound III of embodiment 1, characterized by an XRPD pattern comprising at least ten 2-theta values selected from: 9.53±0.2°, 10.04±0.2°, 11.60±0.2°, 14.57±0.2°, 17.22±0.2°, 17.50±0.2°, 20.04±0.2°, 20.36±0.2°, 22.34±0.2°, 23.73±0.2°, 25.48±0.2°, 26.06±0.2°, 27.38±0.2°, and 32.20±0.2°. 5. Pattern 1 of compound III of embodiment 1, characterized by an XRPD pattern comprising at least nine 2-theta values selected from: 9.53±0.2°, 10.04±0.2°, 11.60±0.2°, 14.57±0.2°, 17.22±0.2°, 17.50±0.2°, 20.04±0.2°, 20.36±0.2°, 22.34±0.2°, 23.73±0.2°, 25.48±0.2°, 26.06±0.2°, 27.38±0.2°, and 32.20±0.2°. 6. Compound III pattern 1 of embodiment 1, characterized by an XRPD pattern comprising at least eight 2-theta values selected from: 6.9.53±0.2°, 10.04±0.2°, 11.60±0.2°, 14.57±0.2°, 17.22±0.2°, 17.50±0.2°, 20.04±0.2°, 20.36±0.2°, 22.34±0.2°, 23.73±0.2°, 25.48±0.2°, 26.06±0.2°, 27.38±0.2°, and 32.20±0.2°. 7. Pattern 1 of compound III of embodiment 1, characterized by an XRPD pattern comprising at least seven 2-theta values selected from: 7.9.53±0.2°, 10.04±0.2°, 11.60±0.2°, 14.57±0.2°, 17.22±0.2°, 17.50±0.2°, 20.04±0.2°, 20.36±0.2°, 22.34±0.2°, 23.73±0.2°, 25.48±0.2°, 26.06±0.2°, 27.38±0.2°, and 32.20±0.2°. 8. Compound III pattern 1 of embodiment 1, characterized by an XRPD pattern comprising at least six 2-theta values selected from: 8.9.53±0.2°, 10.04±0.2°, 11.60±0.2°, 14.57±0.2°, 17.22±0.2°, 17.50±0.2°, 20.04±0.2°, 20.36±0.2°, 22.34±0.2°, 23.73±0.2°, 25.48±0.2°, 26.06±0.2°, 27.38±0.2°, and 32.20±0.2°. 9. Pattern 1 of compound III of embodiment 1, characterized by an XRPD pattern comprising at least five 2-theta values selected from 9.9.53±0.2°, 10.04±0.2°, 11.60±0.2°, 14.57±0.2°, 17.22±0.2°, 17.50±0.2°, 20.04±0.2°, 20.36±0.2°, 22.34±0.2°, 23.73±0.2°, 25.48±0.2°, 26.06±0.2°, 27.38±0.2°, and 32.20±0.2°. 10. In one embodiment, Compound III Pattern 2 has the following peaks: 8.94±0.2°, 9.89±0.2°, 9.91±0.2°, 11.66±0.2°, 12.11±0.2°, 15.13±0.2°, 17.85±0.2°, 18.15±0.2°, 19.90±0.2°, 20.38±0.2°, 22.94±0.2° , 25.09±0.2°, 26.54±0.2°, 26.90±0.2°, 27.38±0.2°, 28.28±0.2°, 28.95±0.2°, 29.64±0.2°, and 38.07±0.2°. 11. Compound III pattern 2 of embodiment 10, characterized by an XRPD pattern comprising at least 12 2-theta values selected from 11.8.94±0.2°, 9.89±0.2°, 9.91±0.2°, 11.66±0.2°, 12.11±0.2°, 15.13±0.2°, 17.85±0.2°, 18.15±0.2°, 19.90±0.2°, 20.38±0.2°, 22.94±0.2°, 25.09±0.2°, 26.54±0.2°, 26.90±0.2°, 27.38±0.2°, 28.28±0.2°, 28.95±0.2°, 29.64±0.2°, and 38.07±0.2°. 11. Compound III Pattern 2 of embodiment 10, characterized by an XRPD pattern comprising at least three, four, five, or six 2-theta values independently selected from 12.8.94±0.2°, 9.89±0.2°, 9.91±0.2°, 11.66±0.2°, 12.11±0.2°, 15.13±0.2°, 17.85±0.2°, 18.15±0.2°, 19.90±0.2°, 25.09±0.2°, 29.64±0.2°, and 38.07±0.2°. 11. Compound III pattern 2 of embodiment 10, characterized by an XRPD pattern comprising at least 11 2-theta values selected from 13.8.94±0.2°, 9.89±0.2°, 9.91±0.2°, 11.66±0.2°, 12.11±0.2°, 15.13±0.2°, 17.85±0.2°, 18.15±0.2°, 19.90±0.2°, 25.09±0.2°, 29.64±0.2°, and 38.07±0.2°. 11. Compound III Pattern 2 of embodiment 10, characterized by an XRPD pattern comprising at least ten 2-theta values selected from 14.8.94±0.2°, 9.89±0.2°, 9.91±0.2°, 11.66±0.2°, 12.11±0.2°, 15.13±0.2°, 17.85±0.2°, 18.15±0.2°, 19.90±0.2°, 25.09±0.2°, 29.64±0.2°, and 38.07±0.2°. 11. Compound III Pattern 2 of embodiment 10, characterized by an XRPD pattern comprising at least nine 2-theta values selected from 15.8.94±0.2°, 9.89±0.2°, 9.91±0.2°, 11.66±0.2°, 12.11±0.2°, 15.13±0.2°, 17.85±0.2°, 18.15±0.2°, 19.90±0.2°, 25.09±0.2°, 29.64±0.2°, and 38.07±0.2°. 11. Compound III pattern 2 of embodiment 10, characterized by an XRPD pattern comprising at least eight 2-theta values selected from 16.8.94±0.2°, 9.89±0.2°, 9.91±0.2°, 11.66±0.2°, 12.11±0.2°, 15.13±0.2°, 17.85±0.2°, 18.15±0.2°, 19.90±0.2°, 25.09±0.2°, 29.64±0.2°, and 38.07±0.2°. 11. Compound III Pattern 2 of embodiment 10, characterized by an XRPD pattern comprising at least seven 2-theta values selected from 17.8.94±0.2°, 9.89±0.2°, 9.91±0.2°, 11.66±0.2°, 12.11±0.2°, 15.13±0.2°, 17.85±0.2°, 18.15±0.2°, 19.90±0.2°, 25.09±0.2°, 29.64±0.2°, and 38.07±0.2°. 11. Compound III pattern 2 of embodiment 10, characterized by an XRPD pattern comprising at least six 2-theta values selected from 18.8.94±0.2°, 9.89±0.2°, 9.91±0.2°, 11.66±0.2°, 12.11±0.2°, 15.13±0.2°, 17.85±0.2°, 18.15±0.2°, 19.90±0.2°, 25.09±0.2°, 29.64±0.2°, and 38.07±0.2°. 11. Compound III pattern 2 of embodiment 10, characterized by an XRPD pattern comprising at least five 2-theta values selected from 19.8.94±0.2°, 9.89±0.2°, 9.91±0.2°, 11.66±0.2°, 12.11±0.2°, 15.13±0.2°, 17.85±0.2°, 18.15±0.2°, 19.90±0.2°, 25.09±0.2°, 29.64±0.2°, and 38.07±0.2°.

[0290] Compound III Pattern 1 can be prepared, for example, by precipitation from isopropanol and heptane (Example 15, Table 40). In certain non-limiting embodiments, Compound III Pattern 1 can be prepared by precipitation from S p Compound I free base is dissolved in isopropanol, e.g., about 100 mg of Sp Compound I was prepared by dissolving about 0.25 mL to about 0.5 mL of isopropanol. To this solution, about 1.0 equivalent of fumaric acid was added, and the mixture was stirred at ambient or elevated temperature, e.g., about 25°C to about 60°C. Next, about 2 to 5 times as much heptane as isopropanol was added. The resulting mixture was stirred at ambient or elevated temperature, e.g., about 25°C to about 60°C, and then slowly cooled (e.g., about 0.01°C / min to 1°C / min), and the solid was isolated by filtration and then dried at ambient or reduced pressure.

[0291] IV. Definition Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. All patents, applications, and published applications mentioned herein are incorporated by reference in their entirety unless otherwise stated. In the event that there are multiple definitions for a term herein, the definition in this section prevails unless otherwise stated.

[0292] As used herein, the abbreviations for any protecting groups, amino acids, and other compounds follow common usage, recognized abbreviations, or the IUPAC-IUB Biochemical Nomenclature Commission (see Biochem. 11:942-944 (1972)) unless otherwise specified.

[0293] As used herein, the term "alcohol solvent" refers to a solvent having free hydroxyl groups that is liquid at either room temperature or the temperature of use. Non-limiting examples of alcohol solvents include methanol, ethanol, ethanediol, isopropanol, n-propanol, glycerol, n-butanol, iso-butanol, tert-butanol, 2-butanol, n-pentanol (n-amyl alcohol), iso-pentanol (isoamyl alcohol), neopentyl alcohol, hexanol, cyclohexanol, cyclohexanediol, phenol, benzyl alcohol, propargyl alcohol, diethylene glycol, 1,2-propanediol, heptanol, octanol, nonanol, and decanol.

[0294] As used herein, "aliphatic solvent" refers to a hydrocarbon-based solvent that is liquid at room temperature or the temperature of use. Non-limiting examples of aliphatic solvents include pentane, isopentane, cyclopentane, n-hexane, hexane (mixture of isomers), cyclohexane, 2-hexene, 3-hexene, methylcyclohexane, heptane, octane, isooctane, petroleum ether, naphtha, mineral spirits, nonane, decane, undecane, and dodecane.

[0295] As used herein, the term "ether solvent" refers to a solvent that contains at least one ether bond and is liquid at room temperature or the temperature of use. Examples of ether solvents include, but are not limited to, diethyl ether, diisopropyl ether, methyl tert-butyl ether, dibutyl ether, tert-amyl ethyl ether, cyclopentyl methyl ether, di-tert-butyl ether, ethyl tert-butyl ether, propylene glycol methyl ether, ethylene glycol ethyl ether, glyme, diglyme, glycol ether, dioxane, tetrahydrofuran, 2-methyltetrahydrofuran, 4-methyltetrahydrofuran, and tetrahydropyran.

[0296] When a range of values is given, it is understood that the upper and lower limits, and each intervening value between the upper and lower limits of the range, are included within the scope of the embodiment.

[0297] As used herein, "about" refers to a range that includes 10% less and up to 10% more than the stated value. For example, "about 100 milligrams" includes all values from 90 milligrams to 110 milligrams.

[0298] The terms "therapeutically effective amount" and "effective amount" are used to refer to an amount of an active compound or pharmaceutical agent, or a metabolite thereof, that elicits a desired therapeutic effect. For example, an effective amount of a compound can be the amount necessary to prevent, alleviate, or ameliorate symptoms of a disease or prolong the survival of a subject, such as a human, being treated. This response can occur in a tissue, system, animal, or human, and includes alleviation of signs or symptoms of the disease being treated.

[0299] isotope substitution The present invention includes, but is not limited to, the use of any of the compounds, pharmaceutical compositions, and active compounds described herein, including, but not limited to, Compound I monofumarate, Compound II, or Compound III, with desired isotopic substitution of atoms that are enriched above the natural abundance of the isotope, i.e., enriched. Isotopes are atoms having the same atomic number but different mass numbers, i.e., the same number of protons but different numbers of neutrons. As a general example and not by way of limitation, deuterium ( 2 H) and tritium ( 3 H) may be used anywhere in the structures depicted. Alternatively, or additionally, isotopes of carbon, such as 13 C and 14 C may also be used. A preferred isotopic substitution is the substitution of deuterium for hydrogen at one or more positions on a molecule to improve drug performance. Deuterium can be attached at the position of bond cleavage during metabolism (α-deuterium kinetic isotope effect) or adjacent to or near the site of bond cleavage (β-deuterium kinetic isotope effect). Achillion Pharmaceuticals, Inc. (WO 2014 / 169278 and WO 2014 / 169280) describes the deuteration of nucleotides to improve their pharmacokinetics or efficacy, including deuteration at the 5-position of the molecule.

[0300] Substitution with isotopes such as deuterium can confer certain therapeutic advantages due to greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements. Substitution of hydrogen with deuterium at a site of metabolic degradation can decrease the rate of metabolism at that bond or eliminate metabolism at that bond. At any position in a compound where a hydrogen atom can be present, the hydrogen atom can be substituted with protium ( 1 H), deuterium ( 2 H) and tritium ( 3 and 1,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56,57,58,59,59,60,61,62,63,64,65,66,67,68,69,70,71,72,74,75,76,77,78,79,71,71,72,71,72,73,74,75,

[0301] The term "isotopically labeled" analogues refers to "deuterated analogues," 13 C-labeled analogs" or "deuterated / 13 The term "deuterated analog" refers to an analog that is a "C-labeled analog." 1 H) is converted to the H-isotope, namely deuterium ( 2 H). Deuterium substitution may be partial or complete. Partial deuterium substitution means that at least one hydrogen is replaced by at least one deuterium. In certain embodiments, the isotope is enriched to 90%, 95%, or 99% or more in the isotope at any desired position. In some embodiments, it is deuterium that is enriched to 90%, 95%, or 99% at the desired position. Unless specified to the contrary, deuteration is at least 80% at the selected position. Deuteration of a nucleoside may occur at any substitutable hydrogen to produce the desired result.

[0302] V. Treatment or Prevention of HPV-Induced Intraepithelial Neoplasia In exemplary, non-limiting embodiments, methods are provided for treating HPV infection or HPV-induced intraepithelial neoplasia, as further described herein, comprising administering an effective amount of one or a combination of active compounds, as described herein, in a topical formulation sufficient to treat the neoplasia or its resulting effects. Types of HPV-induced intraepithelial neoplasia include, but are not limited to, cervical, vaginal, vulvar, penile, perianal, and anal.

[0303] In exemplary embodiments, the formulation for treating intraepithelial neoplasia is a dosage form containing about 0.005 mg to about 50 mg, about 0.05 mg to about 40 mg, about 0.1 mg to about 30 mg, about 0.5 mg to about 20 mg, about 1 mg to about 20 mg, about 1 mg to about 15 mg, or about 1 mg to about 10 mg of any of the active compounds described herein, including, but not limited to, Compound I monofumarate, Compound II, or Compound III. In certain embodiments, the formulation for treating intraepithelial neoplasia is a dosage form containing about 0.01 mg to about 10 mg, about 0.05 mg to about 5 mg, about 0.05 mg to about 0.15 mg, about 0.15 mg to about 0.45 mg, or about 0.5 mg to about 1.5 mg of any of the active compounds described herein, including, but not limited to, Compound I monofumarate, Compound II, or Compound III. In certain embodiments, the formulation for treating intraepithelial neoplasia is a dosage form containing about or at least 0.005 mg, 0.01 mg, 0.03 mg, 0.05 mg, 0.1 mg, 0.3 mg, 0.5 mg, 0.7 mg, 1 mg, 1.5 mg, 2 mg, 2.5 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, or 50 mg of Compound I monofumarate, Compound II, or Compound III.

[0304] In certain embodiments, the formulation for treating intraepithelial neoplasia is in a dosage form containing about 0.001 mg to about 20 mg, about 0.005 mg to about 10 mg, about 0.01 mg to about 5 mg, about 0.03 mg to about 1 mg, or about 0.05 mg to about 0.3 mg of Compound I monofumarate, Compound II, or Compound III.

[0305] In certain embodiments, the topical formulation is administered twice a day, once a day, or several days a week (such as two or three days a week) as needed to achieve the desired results. In certain embodiments, the topical formulation is administered on a weekly schedule for one, two, three, four, five, six or more weeks. In certain aspects, the topical formulation is administered on a three times a week schedule for two, three, four, five, or six weeks.

[0306] In certain embodiments, the compound can be administered in one or more treatment cycles, including a treatment cycle and a rest cycle, where a treatment cycle comprises administering a compound described herein, followed by a rest cycle (including a period of no treatment) before the next treatment cycle. In certain embodiments, the rest cycle is from about 1 day to about 6 months. In certain embodiments, the rest cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, or more before the next treatment cycle. In certain embodiments, multiple treatment cycles are applied, for example, 1, 2, 3, 4, 5, or 6 treatment cycles.

[0307] Dosage forms that do not adhere to the target site may be removed, which may interfere with treatment. Dosage forms that adhere to the target site and dissolve rapidly with low flow rates have been discovered. Adhesion to the target site may also prevent exposure to healthy tissue, limiting toxicity and side effects. Dosage forms that soften, degrade, and / or disintegrate rapidly with low flow rates are advantageous for causing rapid release of the active compound to the target tissue. For example, dosage forms that disintegrate with fluid volumes of less than about 50 μL, less than about 100 μL, less than about 125 μL, less than about 150 μL, less than about 175 μL, less than about 200 μL, or less than about 250 μL particularly facilitate penetration of the drug into the target site.

[0308] In certain embodiments, the dosage form is a gel. In certain embodiments, the dosage form is a cream. In certain embodiments, the dosage form is a tablet. In certain embodiments, the dosage form disintegrates in about 1 second to about 10 seconds. In certain embodiments, the dosage form disintegrates in about 10 seconds to 1 minute. In certain embodiments, the dosage form disintegrates in about 1 minute to about 1 hour. In certain embodiments, the dosage form disintegrates in about 1 hour to 6 hours.

[0309] The physical dimensions of a dosage form can affect its effectiveness. Thinner tablets have a higher surface area to volume ratio, which can lead to faster disintegration and better coverage of the target area. In certain embodiments, the dosage form is less than about 6 millimeters, 5 millimeters, 4 millimeters, 3 millimeters, or 2 millimeters thick in its smallest dimension.

[0310] The formulation of the dosage form is important for proper administration of the active agent to intraepithelial tissues. For example, the formulation can be prepared for use as a tablet, a reconstituted powder, a dry powder, a semi-solid dosage form, a film, or a pessary (i.e., a vaginal suppository).

[0311] Some embodiments disclosed herein include the use of an effective amount of Compound I monofumarate, Compound II, or Compound III in the manufacture of a medicament for ameliorating or treating a human papillomavirus infection, which can be ameliorated or treated by inhibiting viral replication through inhibiting viral DNA synthesis. Other embodiments disclosed herein include the use of an effective amount of any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, to ameliorate or treat a human papillomavirus infection, which can be ameliorated or treated by inhibiting viral replication through inhibiting viral DNA synthesis.

[0312] Certain non-limiting embodiments disclosed herein include methods for ameliorating or treating human papillomavirus infection, which may include contacting cells infected with human papillomavirus in a subject with an effective amount of any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, wherein the infection is ameliorated or treated by inhibiting viral DNA synthesis. Further, still other embodiments disclosed herein include methods for ameliorating or treating human papillomavirus infection, which may include administering an effective amount of Compound I monofumarate, Compound II, or Compound III to a subject infected with human papillomavirus, wherein the human papillomavirus infection can be ameliorated or treated by inhibiting viral replication through inhibiting viral DNA synthesis. Some embodiments disclosed herein relate to Compound I monofumarate, Compound II, or Compound III for use in ameliorating or treating human papillomavirus infection, wherein the human papillomavirus infection can be ameliorated or treated by inhibiting viral replication through inhibiting viral DNA synthesis.

[0313] In some embodiments, the human papillomavirus can be a high-risk human papillomavirus, such as those described herein. For example, the high-risk human papillomavirus can be selected from HPV-16, HPV-18, HPV-31, HPV-33, HPV-35, HPV-39, HPV-45, HPV-51, HPV-52, HPV-56, HPV-58, HPV-59, HPV-68, HPV-73, and HPV-82. In some embodiments, the human papillomavirus can be HPV-16. In some embodiments, the human papillomavirus can be HPV-11. In some embodiments, the human papillomavirus can be HPV-18. In some embodiments, the human papillomavirus may be one or more of the following high-risk types: HPV-31, HPV-33, HPV-35, HPV-39, HPV-45, HPV-51, HPV-52, HPV-56, HPV-58, HPV-59, HPV-68, HPV-73, and HPV-82. As described herein, the presence of HPV infection can be detected using a Papanicolaou test (Pap smear) and / or a DNA probe test (e.g., an HPV DNA probe test for one or more high-risk HPV types). Thus, in some embodiments, an effective amount of Compound I monofumarate, Compound II, or Compound III can be provided to a subject diagnosed with an HPV infection, e.g., a high-risk HPV infection, by a DNA test. In some embodiments, an effective amount of Compound I monofumarate, Compound II, or Compound III can be provided to a subject diagnosed with an HPV infection or a disease associated with an HPV infection, as identified by a Papanicolaou test.

[0314] In certain embodiments, an effective amount of Compound I monofumarate, Compound II, or Compound III may be provided to a subject with a Papanicolaou test result that does not indicate that the disease has progressed to cervical cancer. The Bethesda System is a standardized scoring system for reporting Pap smear results, assigning grades 1 to 3 based on severity. Grade 1 CIN (CIN1) indicates mild dysplasia. Grades 2 and 3 CIN (CIN2, CIN3) are more severe and typically require intervention. In certain embodiments, Compound I monofumarate, Compound II, or Compound III is used to treat CIN1 (grade 1 cervical intraepithelial neoplasia). In certain embodiments, Compound I monofumarate, Compound II, or Compound III is used to treat CIN2 (grade 2 cervical intraepithelial neoplasia). In certain embodiments, Compound I monofumarate, Compound II, or Compound III is used to treat CIN3 (grade 3 cervical intraepithelial neoplasia).

[0315] In certain embodiments, a pharmaceutical composition comprising Compound I monofumarate, Compound II, or Compound III is used in the manufacture of a medicament for the treatment of CIN1 (grade 1 cervical intraepithelial neoplasia). In certain embodiments, a pharmaceutical composition comprising Compound I monofumarate, Compound II, or Compound III is used in the manufacture of a medicament for the treatment of CIN2 (grade 2 cervical intraepithelial neoplasia). In certain embodiments, a pharmaceutical composition comprising Compound I monofumarate, Compound II, or Compound III is used in the manufacture of a medicament for the treatment of CIN3 (grade 3 cervical intraepithelial neoplasia).

[0316] In certain embodiments, Compound I monofumarate, Compound II, or Compound III, optionally in a pharmaceutically acceptable carrier, is used to treat a condition selected from the group consisting of atypical squamous cells of undetermined significance (ASC-US), atypical glandular cells (AGC), low-grade squamous intraepithelial lesion (LSIL), atypical squamous cells (high-grade squamous intraepithelial lesion cannot be excluded) (ASC-H), high-grade squamous intraepithelial lesion (HSIL), adenocarcinoma in situ (AIS), and cervical cancer (e.g., squamous cell carcinoma or adenocarcinoma).

[0317] In certain embodiments, an effective amount of Compound II may be provided to a subject whose Papanicolaou test result does not indicate that the disease has progressed to cervical cancer. In certain embodiments, Compound II is used to treat CIN1 (grade 1 cervical intraepithelial neoplasia). In certain embodiments, Compound II is used to treat CIN2 (grade 2 cervical intraepithelial neoplasia). In certain embodiments, Compound II is used to treat CIN3 (grade 3 cervical intraepithelial neoplasia).

[0318] In certain embodiments, a pharmaceutical composition comprising Compound II is used in the manufacture of a medicament for the treatment of CIN1 (grade 1 cervical intraepithelial neoplasia). In certain embodiments, a pharmaceutical composition comprising Compound II is used in the manufacture of a medicament for the treatment of CIN2 (grade 2 cervical intraepithelial neoplasia). In certain embodiments, a pharmaceutical composition comprising Compound II is used in the manufacture of a medicament for the treatment of CIN3 (grade 3 cervical intraepithelial neoplasia).

[0319] In certain embodiments, Compound II, optionally in a pharmaceutically acceptable carrier, is used to treat a condition selected from the group consisting of atypical squamous cells of undetermined significance (ASC-US), atypical glandular cells (AGC), low-grade squamous intraepithelial lesion (LSIL), atypical squamous cells (high-grade squamous intraepithelial lesion cannot be excluded) (ASC-H), high-grade squamous intraepithelial lesion (HSIL), adenocarcinoma in situ (AIS), and cervical cancer (squamous cell carcinoma or adenocarcinoma).

[0320] In certain embodiments, Compound I monofumarate, Compound II, or Compound III is used in the manufacture of a medicament for the treatment of anal intraepithelial neoplasia. In certain embodiments, Compound I monofumarate, Compound II, or Compound III is used in the manufacture of a medicament for the treatment of perianal intraepithelial neoplasia. In certain embodiments, Compound I monofumarate, Compound II, or Compound III is used in the manufacture of a medicament for the treatment of vulvar intraepithelial neoplasia. In certain embodiments, Compound I monofumarate, Compound II, or Compound III is used in the manufacture of a medicament for the treatment of penile intraepithelial neoplasia. In certain embodiments, Compound I monofumarate, Compound II, or Compound III is used in the manufacture of a medicament for the treatment of vaginal intraepithelial neoplasia.

[0321] In certain embodiments, Compound I monofumarate, Compound II, or Compound III is used to treat anal intraepithelial neoplasia. In certain embodiments, Compound I monofumarate, Compound II, or Compound III is used to treat perianal intraepithelial neoplasia. In certain embodiments, Compound I monofumarate, Compound II, or Compound III is used to treat vulvar intraepithelial neoplasia. In certain embodiments, Compound I monofumarate, Compound II, or Compound III is used to treat penile intraepithelial neoplasia. In certain embodiments, Compound I monofumarate, Compound II, or Compound III is used to treat vaginal intraepithelial neoplasia.

[0322] In some embodiments, the human papillomavirus can be a low-risk human papillomavirus, including those described herein. In some embodiments, the human papillomavirus can be HPV-6. In some embodiments, the human papillomavirus can be HPV-11.

[0323] Compound I monofumarate, Compound II, or Compound III can be used to ameliorate and / or treat infections caused by one or more types of human papillomavirus. For example, Compound I monofumarate, Compound II, or Compound III can be used to ameliorate and / or treat infections caused by HPV-16 and / or HPV-18. In certain embodiments, Compound I monofumarate, Compound II, or Compound III can be used to treat high-risk HPV infections. In certain embodiments, Compound I monofumarate, Compound II, or Compound III can be used to treat related diseases or conditions resulting from high-risk HPV infections. In some embodiments, Compound I monofumarate, Compound II, or Compound III can be used to ameliorate and / or treat infections involving both high-risk and low-risk HPV.

[0324] Compound I monofumarate, Compound II, or Compound III can be used in the manufacture of a medicament used to ameliorate and / or treat infections caused by one or more types of human papillomavirus. For example, Compound I monofumarate, Compound II, or Compound III can be used in the manufacture of a medicament used to ameliorate and / or treat infections caused by HPV-16 and / or HPV-18. In certain embodiments, Compound I monofumarate, Compound II, or Compound III can be used in the manufacture of a medicament used to treat high-risk HPV infections. In certain embodiments, Compound I monofumarate, Compound II, or Compound III can be used in the manufacture of a medicament used to treat related diseases or conditions resulting from high-risk HPV infections. In some embodiments, Compound I monofumarate, Compound II, or Compound III can be used in the manufacture of a medicament used to ameliorate and / or treat infections involving both high-risk and low-risk HPV.

[0325] As will be readily apparent to those skilled in the art, useful in vivo dosages and the particular mode of administration will vary depending on the age, weight, severity of the affliction, the particular compounds used, and the particular application for which the compounds are used. The determination of effective dosage levels, i.e., the dosage levels necessary to achieve the desired result, can be accomplished by those skilled in the art using routine methods, such as human clinical trials and in vitro studies.

[0326] In certain embodiments, a pharmaceutical composition comprising Compound II is used to treat a condition associated with or resulting from exposure to or infection with HPV. In certain embodiments, a pharmaceutical composition comprising Compound II is used to treat precancerous cervical lesions. In certain embodiments, a pharmaceutical composition comprising Compound II is used to treat cervical intraepithelial neoplasia. In certain embodiments, a pharmaceutical composition comprising Compound II is used to treat vaginal and anal intraepithelial neoplasia. In certain embodiments, a pharmaceutical composition comprising Compound II is used to treat cervical cancer. In certain embodiments, a pharmaceutical composition comprising Compound II is used to treat rectal cancer. In certain embodiments, a pharmaceutical composition comprising Compound II is used to treat penile cancer. In certain embodiments, a pharmaceutical composition comprising Compound II is used to treat vaginal cancer. In certain aspects, a pharmaceutical composition comprising Compound II is used to treat oropharyngeal cancer.

[0327] In certain embodiments, a pharmaceutical composition comprising Compound II is used in the manufacture of a medicament for the treatment of a condition associated with or resulting from exposure to or infection with HPV. In certain embodiments, a pharmaceutical composition comprising Compound II is used in the manufacture of a medicament for the treatment of precancerous cervical lesions. In certain embodiments, a pharmaceutical composition comprising Compound II is used in the manufacture of a medicament for the treatment of cervical intraepithelial neoplasia. In certain embodiments, a pharmaceutical composition comprising Compound II is used in the manufacture of a medicament for the treatment of vaginal and anal intraepithelial neoplasia. In certain embodiments, a pharmaceutical composition comprising Compound II is used in the manufacture of a medicament for the treatment of cervical cancer. In certain embodiments, a pharmaceutical composition comprising Compound II is used in the manufacture of a medicament for the treatment of rectal cancer. In certain embodiments, a pharmaceutical composition comprising Compound II is used in the manufacture of a medicament for the treatment of penile cancer. In certain embodiments, a pharmaceutical composition comprising Compound II is used in the manufacture of a medicament for the treatment of vaginal cancer. In certain embodiments, a pharmaceutical composition comprising Compound II is used in the manufacture of a medicament for the treatment of oropharyngeal cancer.

[0328] It is advantageous for the dosage form to be easily applied to the target site. Direct application to the target site prevents systemic exposure and toxicity. To place the dosage form at the target site, the dosage form may be applied by an applicator. In certain embodiments, the dosage form is applied by a vaginal applicator. In certain embodiments, the dosage form is applied without an applicator. In certain embodiments, an additional fluid (e.g., a lubricant) is delivered with the dosage form, applied to the dosage form, or applied to the target site or surrounding tissue.

[0329] In certain embodiments, a lubricating fluid is administered in combination with the dosage form to enhance coverage of the cervix, vagina, vulva, anus, perianal area, or penis. In certain embodiments, water is used as the fluid administered with the dosage form. In certain embodiments, a lubricating glycerol- or hydroxyethylcellulose-based water-soluble fluid is used in combination with the dosage form. In certain embodiments, the dosage form is administered without additional fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in less than about 5 milliliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in less than about 4 milliliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in less than about 3 milliliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in less than about 2 milliliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in less than about 1 milliliter of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in less than about 0.75 milliliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in less than about 0.5 milliliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in less than about 0.25 milliliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in less than about 0.2 milliliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in less than about 0.15 milliliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in less than about 0.125 milliliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in less than about 0.1 milliliters of fluid.

[0330] In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in about 10 microliters to about 100 microliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in about 75 microliters to about 250 microliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in about 200 microliters to about 500 microliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in about 400 microliters to about 750 microliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in about 700 microliters to about 1000 microliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in about 1 milliliter to about 2 milliliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in about 2 milliliters to about 3 milliliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in about 3 milliliters to about 4 milliliters of fluid. In certain embodiments, the dosage form softens, disintegrates, and / or dissolves in about 4 milliliters to about 5 milliliters of fluid.

[0331] In certain embodiments, Compound II is administered for at least 1 day, 2 days, 3 days, 4 days, 5 days, or 6 days, either continuously or non-continuously.

[0332] In certain embodiments, Compound II is administered once a week. In certain embodiments, Compound II is administered once a week for up to 12 weeks. In certain embodiments, Compound II is administered once a week for up to 10 weeks. In certain embodiments, Compound II is administered once a week for up to 8 weeks. In certain embodiments, Compound II is administered once a week for up to 6 weeks. In certain embodiments, Compound II is administered once a week for up to 4 weeks. In certain embodiments, Compound II is administered once a week for up to 2 weeks. In certain embodiments, Compound II is administered once a week for up to 1 week.

[0333] In certain embodiments, Compound II is administered twice a week. In certain embodiments, Compound II is administered twice a week for up to 12 weeks. In certain embodiments, Compound II is administered twice a week for up to 10 weeks. In certain embodiments, Compound II is administered twice a week for up to 8 weeks. In certain embodiments, Compound II is administered twice a week for up to 6 weeks. In certain embodiments, Compound II is administered twice a week for up to 4 weeks. In certain embodiments, Compound II is administered twice a week for up to 2 weeks. In certain embodiments, Compound II is administered twice a week for up to 1 week.

[0334] In certain embodiments, Compound II is administered three times per week. In certain embodiments, Compound II is administered three times per week for up to 12 weeks. In certain embodiments, Compound II is administered three times per week for up to 10 weeks. In certain embodiments, Compound II is administered three times per week for up to 8 weeks. In certain embodiments, Compound II is administered three times per week for up to 6 weeks. In certain embodiments, Compound II is administered three times per week for up to 4 weeks. In certain embodiments, Compound II is administered three times per week for up to 2 weeks. In certain embodiments, Compound II is administered three times per week for up to 1 week.

[0335] In certain embodiments, Compound II is administered daily. In certain embodiments, Compound II is administered daily for up to 12 weeks or indefinitely, as directed by a healthcare professional. In certain embodiments, Compound II is administered daily for up to 10 weeks. In certain embodiments, Compound II is administered daily for up to 8 weeks. In certain embodiments, Compound II is administered daily for up to 6 weeks. In certain embodiments, Compound II is administered daily for up to 4 weeks. In certain embodiments, Compound II is administered daily for up to 2 weeks. In certain embodiments, Compound II is administered daily for up to 1 week. In certain embodiments, about 0.05 mg to about 0.3 mg of Compound II is administered daily for 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, or more, as directed by a healthcare professional.

[0336] In certain embodiments, compound I monofumarate is administered three times per week. In certain embodiments, compound I monofumarate is administered three times per week for up to 12 weeks. In certain embodiments, compound I monofumarate is administered three times per week for up to 10 weeks. In certain embodiments, compound I monofumarate is administered three times per week for up to 8 weeks. In certain embodiments, compound I monofumarate is administered three times per week for up to 6 weeks. In certain embodiments, compound I monofumarate is administered three times per week for up to 4 weeks. In certain embodiments, compound I monofumarate is administered three times per week for up to 2 weeks. In certain embodiments, compound I monofumarate is administered three times per week for up to 1 week.

[0337] In certain embodiments, compound I monofumarate is administered daily. In certain embodiments, compound I monofumarate is administered daily for up to 12 weeks or indefinitely as directed by a healthcare professional. In certain embodiments, compound I monofumarate is administered daily for up to 10 weeks. In certain embodiments, compound I monofumarate is administered daily for up to 8 weeks. In certain embodiments, compound I monofumarate is administered daily for up to 6 weeks. In certain embodiments, compound I monofumarate is administered daily for up to 4 weeks. In certain embodiments, compound I monofumarate is administered daily for up to 2 weeks. In certain embodiments, compound I monofumarate is administered daily for up to 1 week.

[0338] In certain embodiments, Compound I monofumarate, Compound II, or Compound III may be administered three, four, five, or six times per week. In certain embodiments, Compound I monofumarate, Compound II, or Compound III may be administered once per day. In certain embodiments, Compound I monofumarate, Compound II, or Compound III may be administered twice per day. In certain embodiments, Compound I monofumarate, Compound II, or Compound III may be administered three, four, or more times per day. In certain embodiments, Compound I monofumarate, Compound II, or Compound III may be administered daily.

[0339] In certain embodiments, the compound can be administered in one or more treatment cycles, including a treatment cycle and a rest cycle, where a treatment cycle comprises administering a compound described herein, followed by a rest cycle (including a period of no treatment) before the next treatment cycle. In certain embodiments, the rest cycle is from about 1 day to about 6 months. In certain embodiments, the rest cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, or more before the next treatment cycle. In certain embodiments, multiple treatment cycles are applied, for example, 1, 2, 3, 4, 5, or 6 treatment cycles.

[0340] As mentioned above, many compounds are being investigated for the treatment of HPV-induced tumors, but none have yet been approved. For non-limiting examples of investigated approaches, see Ahn WS, et al. Protective effects of green tea extracts (polyphenon E and EGCG) on human cervical lesions. Eur. J. Cancer Prev. 2003;12:383-390; Ashrafian L, et al. Double-blind randomized placebo-controlled multicenter clinical trial (phase IIa) on diindolymethane's efficacy and safety in the treatment of CIN: implications for cervical cancer prevention. EPMA J. 2015;6:doi: 10.1186 / s13167-13015-10048-13169; Bossens M., et al. Safety and tolerance of cidofovir as a 2% gel for local application in high-grade cervical intraepithelial neoplasia: A phase 1 investigation. Int. J. Clin. Pharmacol. 2018;56:134-141, Chen FP Efficacy of imiquimod 5% cream for persistent human papillomavirus in genital intraepithelial neoplasm. Taiwanese J. Obstetrics Gynecol. 2013;52(4):475-478, Choo Y., et al. Intravaginal application of leukocyte interferon gel in the treatment of cervical intraepithelial neoplasia (CIN) Arch Gynecol.1985;237:51-54、de Witte C.J et al. Imiquimod in cervical, vaginal and vulvar intraepithelial neoplasia: a review. Gynecol. Oncol. 2015;139:377-384、Desravines N, et al. Low dose 5-fluorouracil intravaginal therapy for the treatment of cervical intraepithelial neoplasia 2 / 3: A case series. J. Gynecol. Surg. 2020;36、DiSilvestro P.A., et al. Treatment of cervical intraepithelial neoplasia levels 2 and 3 with adapalene, a retinoid-related molecule. J. Low Genit Tract. Dis. 2001;5:33-37、Graham V., et al. Phase II trial of beta-all-transretinoic acid for cervical intraepithelial neoplasia via a collagen sponge and cervical cap. West. J. Med. 1986;145:192-195、Grimm C., et al. Treatment of cervical intraepithelial neoplasia with topical imiquimod: a randomized controlled trial. Obstet. Gynecol. 2012;120(1):152-159、Hampson L., et al. A single-arm, proof-of-concept trial of lopimune (lopinavir / ritonavir) as treatment for HPV-related pre-invasive cervical disease. PLoS ONE. 2016;11、Helm C.W.et al. Retinoids for preventing the progression of cervical intra-epithelial neoplasia. Cochrane Systematic Review. 2013、Hubert P., et al. Local applications of GM-CSF induce the recruitment of immune cells in cervical low-grade squamous intraepithelial lesions. Am. J. Reprod. Immunol. 2010;64:126-136、Koeneman MM, et al. Topical Imiquimod treatment of high-grade Cervical intraepithelial neoplasia (TOPIC trial): study protocol for a randomized controlled trial. BMC Cancer. 2016:doi: 10.1186 / s12885-12016-12187-12883、Krause S., et al. Interferon and cervical dysplasia: CIN III treated with local interferon application. Colposcopy Gynecologic Laser Surgery. 1987;3:195-198、Krebs H.B., et al. Chronic ulcerations following topical therapy with 5-fluorouracil for vaginal human papillomavirus-associated lesions. Obstet. Gynecol. 1991;78(2):205-208、Laccetta G. et al.Effect of the treatment with beta-glucan in women with cervical cytologic report of atypical squamous cells of undetermined significance (ASCUS) and low-grade intraepithelial lesions (L-SIL) Minerva Ginecol. 2015;67:113-120、Meyskens F.L., et al. A phase I trial of beta-all-transretinoic acid delivered via a collagen sponge and a cervical cap for mild or moderate intraepithelial cervical neoplasia. J. Natl Cancer Inst. 1983;71:921-925、Niwa K., et al. Topical vidarabine of 5-fluoruracil treatment against persistent HPV in genital (pre)cancerous lesions. Oncol Reports. 2003;10:1437-1441、Pachman DR, et al. Randomized clinical trial of imiquimod: an adjunct to treating cervical dysplasia. Am. J. Obstet. Gynecol. 2012;206(1):42 e41-47、Rahangdale L et al. Topical 5-fluorouracil for treatment of Cervical Intraepithelial Neoplasia 2: a randomized controlled trial. Am. J. Obstet. Gynecol. 2014;210:e1-e8、Schneider A., et al.Efficacy trial of topically administered Interferon gamma-1beta gel in comparison to laser treatment in cervical intraepithelial neoplasia. Arch. Gynecol Obste. 1995;256:75-83、Silman F.H., et al. 5-fluorouracil / chemosurgery for intraepithelial neoplasia of the lower genital tract. Obstet. Gynecol. 1981;58:356-360、Snoeck R., Noel J.C., Muller C., Clercq De, Bossens M. Cidofovir, a new approach for the treatment of cervix intraepithelial neoplasia III (CIN III) J. Med. Virol. 2000;60:205-209、Stentella P., Biamonti A., Carraro C. Efficacy of carboxymethyl beta-glucan in cervical intraepithelial neoplasia: a retrospective, case-control study. Minerva Ginecol. 2017;69:425-430、Suh-Burgmann E., Sivret J., Duska L.R., Del Carmen M., Seiden M.V. Long-term administration of intravaginal dehydroepiandrosterone on regression of low-grade cervical dysplasia - a pilot study. Gynecol. Obstet. Invest. 2003;55:25-31、Valencia M.H., Pacheco A.C., Quijano T.H., Giron A.V., Lopez C.V.Clinical response to glycyrrhizinic acid in genital infection due to human papillomavirus and low-grade squamous intraepithelial lesion. Clin. Pract. 2011 1(e93), van de Sande A., Koeneman M., Gerestein C., Kruse A., van Kemenade F., van Beekhuizen H. Topical Imiquimod treatment of residual or recurrent cervical intraepithelial neoplasia (TOPIC-2 trial): a study protocol for a randomized controlled trial. BMC Cancer. 2018;18:4510-4517 and Van Pachterbeke C., Bucella D., Rozenberg S. Topical treatment of CIN 2+ by cidofovir: Results of a phase II, double-blind, prospective, placebo-controlled study. Gynecol Onc. 2009;115:69-74. .

[0341] VI. Pharmaceutical Compositions and Dosage Forms In an aspect of the present invention, a pharmaceutical composition according to the present invention comprises an anti-HPV effective amount of any active compound described herein, including, but not limited to, Compound I monofumarate, Compound II, or Compound III described herein, optionally in combination with a pharmaceutically acceptable carrier, additive, or excipient, and / or in combination or alternation with at least one other active compound. In one embodiment, the present invention comprises a solid dosage form of Compound II in a pharmaceutically acceptable carrier. In certain embodiments, the pharmaceutical composition is administered directly to the cervix, vagina, vulva, perianal area, anus, or penis. In certain embodiments, the dosage form is attached to the cervix, vagina, vulva, perianal area, anus, or penis.

[0342] In an aspect of the present invention, a pharmaceutical composition according to the present invention comprises an anti-HPV effective amount of Compound II described herein, optionally in combination with a pharmaceutically acceptable carrier, additive, or excipient, and optionally further in combination with at least one other anti-tumor or anti-viral agent, such as an anti-HPV agent. In certain embodiments, the pharmaceutical composition comprises Compound II in combination with a second anti-viral agent. In certain embodiments, the pharmaceutical composition comprises Compound II in combination with an anti-cancer agent.

[0343] The present invention includes pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III of the present invention, in a pharmaceutically acceptable carrier or excipient in an amount effective to treat HPV infection. In an alternative embodiment, the present invention includes pharmaceutical compositions comprising Compound I monofumarate or Compound II, or a prodrug, of the present invention, in a pharmaceutically acceptable carrier or excipient in an amount effective to prevent HPV infection.

[0344] Those skilled in the art will recognize that a therapeutically effective amount will vary depending on the infection or condition being treated, its severity, the treatment regimen employed, the pharmacokinetics of the drugs used, as well as the patient or subject (animal or human) being treated, and that such a therapeutic amount can be determined by the attending physician or specialist.

[0345] Compound I monofumarate, Compound II or Compound III, or any of the active compounds described herein in accordance with the present invention, can be formulated in a mixture with a pharmaceutically acceptable carrier. For treating HPV infection, the pharmaceutical composition is preferably administered directly to the cervix, vagina, vulva, perianal area, anus, or penis.

[0346] In certain pharmaceutical dosage forms, prodrug forms of the compounds of the present invention can be used to achieve the desired effect, including in particular acylated (acetylated or otherwise) and ether (alkyl and related) derivatives, phosphate esters, thiophosphonamidates, phosphonamidates, and various salt forms of the compounds of the present invention.Those skilled in the art will recognize how to easily convert the compounds of the present invention into prodrug forms to facilitate the delivery of the active compound to the target site of the host organism or patient.Those skilled in the art will also utilize the advantageous pharmacokinetic parameters of the prodrug forms, if applicable, in the delivery of the compound to the target site of the host organism or patient to maximize the intended effect of the compound.

[0347] The amount of any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, included within the therapeutically active formulations according to the present invention is an amount effective to achieve a desired outcome according to the present invention, for example, to treat HPV infection, reduce the likelihood of HCV infection, or inhibit, reduce, and / or eliminate HPV or its secondary effects, including pathologies, conditions, and / or complications that occur secondary to HPV infection.

[0348] Often, dosage forms containing any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, are administered in an amount ranging from about 0.001 milligrams to about 100 milligrams to treat, prevent, or delay the onset of these infections and / or reduce the likelihood of HPV viral infection or secondary pathologies, conditions, or complications of HPV. In certain embodiments, the solid dosage form contains about 0.001 milligrams to about 0.005 milligrams, about 0.005 milligrams to about 0.01 milligrams, about 0.01 milligrams to about 0.03 milligrams, about 0.03 milligrams to about 0.25 milligrams, about 0.20 milligrams to about 0.5 milligrams, about 0.4 milligrams to about 1 milligram, about 0.75 milligrams to about 3 milligrams, about 1 milligram to about 10 milligrams, or about 5 milligrams to about 20 milligrams. In certain embodiments, the solid dosage form comprises at least about 0.001 milligrams, 0.003 milligrams, 0.005 milligrams, 0.01 milligrams, 0.02 milligrams, 0.03 milligrams, 0.04 milligrams, 0.05 milligrams, 0.06 milligrams, 0.07 milligrams, 0.08 milligrams, 0.09 milligrams, 0.1 milligrams, 0.2 milligrams, 0.3 milligrams, 0.4 milligrams, 0.5 milligrams, 0.6 milligrams, 0.7 milligrams, 0.8 milligrams, 0.9 milligrams, 1.0 milligrams, 1.5 milligrams, 2.0 milligrams, 2.5 milligrams, 3 milligrams, 4 milligrams, 5 milligrams, 10 milligrams, 20 milligrams, 30 milligrams, 40 milligrams, or 50 milligrams or more of any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III.

[0349] In certain embodiments, to treat or delay the onset of these infections and / or reduce the likelihood of HPV viral infection or secondary HPV pathologies, conditions, or complications, dosage forms containing any of the active compounds described herein, including, but not limited to, Compound I monofumarate, Compound II, or Compound III, are administered in amounts ranging from about 0.001 mg to about 20 mg, about 0.005 mg to about 10 mg, about 0.01 mg to about 5 mg, about 0.03 mg to about 1 mg, or about 0.05 mg to about 0.3 mg of Compound I monofumarate, Compound II, or Compound III. Dosages typically range from 0.05 mg to 0.3 mg of Compound I monofumarate, Compound II, or Compound III, and can be administered once, twice, three times, or more times per week, up to daily.

[0350] In certain embodiments, to treat infection with high-risk strains of HPV, a dosage form containing any of the active compounds described herein, including, but not limited to, Compound I monofumarate, Compound II, or Compound III, is administered in an amount ranging from about 0.001 mg to about 20 mg, about 0.005 mg to about 10 mg, about 0.01 mg to about 5 mg, about 0.03 mg to about 1 mg, or about 0.05 mg to about 0.3 mg of Compound I monofumarate, Compound II, or Compound III. Dosages for treating high-risk strains of HPV typically range from 0.05 mg to 0.3 mg of Compound I monofumarate, Compound II, or Compound III, and can be administered once, twice, three times, or more times per week, up to daily.

[0351] In certain embodiments, Compound I monofumarate, Compound II, or Compound III may be administered in a gel. In certain embodiments, the gel contains about 0.001% to about 10%, about 0.01% to about 10%, about 0.05% to about 5%, about 0.1% to about 3%, or about 0.1% to about 2% Compound I monofumarate, Compound II, or Compound III (weight / weight). In certain embodiments, the gel contains about 0.001% to about 0.05% Compound I monofumarate, Compound II, or Compound III. In certain embodiments, the gel contains about 0.01% to about 0.5% Compound I monofumarate, Compound II, or Compound III. In certain embodiments, the gel contains about 0.1% to about 5% Compound I monofumarate, Compound II, or Compound III.

[0352] In certain non-limiting embodiments, any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, is administered topically. More generally, Compound I monofumarate, Compound II, or Compound III can be administered in the form of a tablet, capsule, suspension, liquid, emulsion, implant, particle, sphere, cream, ointment, suppository, pessary, transdermal form, gel, mucosal, etc. The dosage form can also be a bilayer tablet in which the entire dose of the active compound is released in one direction (e.g., toward the target tissue).

[0353] In certain embodiments, the dosage form is capable of softening, disintegrating, and / or releasing drug at a low flow rate. In certain embodiments, the dosage form softens and begins to release drug immediately. In certain embodiments, the dosage form softens and begins to release drug gradually. In certain embodiments, the dosage form softens and begins to release drug within 1 hour. In certain embodiments, the dosage form softens and begins to release drug within 2 hours. The dosage form may be formulated to maximize surface area, facilitating disintegration. In certain embodiments, the dosage form is a round tablet. In certain embodiments, the dosage form is an oval tablet. In certain embodiments, the dosage form is a caplet. The width of the tablet is its largest dimension, and the thickness of the tablet is its smaller dimension. In certain embodiments, the dosage form is twice as wide as it is thick. In certain embodiments, the dosage form is three times as wide as it is thick. In certain embodiments, the dosage form is four or more times as wide as it is thick. In certain embodiments, the dosage form is about 0.1 mm to about 5 mm thick. In certain embodiments, the dosage form is about 1 mm to about 2 mm thick. In certain embodiments, the dosage form is about 2 mm to about 3 mm thick. In certain embodiments, the dosage form is about 3 mm to about 4 mm thick. In certain embodiments, the dosage form is about 4 mm to about 5 mm thick. In certain embodiments, the tablet is about 5 mm to about 15 mm thick. In certain embodiments, the dosage form is less than 5 grams. In certain embodiments, the dosage form is about 0.05 grams to about 0.15 grams. In certain embodiments, the dosage form is about 0.1 grams to about 1 gram. In certain embodiments, the dosage form is about 0.75 grams to about 2 grams. In certain embodiments, the dosage form is about 1 gram to about 5 grams.

[0354] In certain embodiments, the dosage form is not easily removed, detached, or displaced from the target site. These desirable properties can be achieved by including a mucoadhesive polymer in the pharmaceutical composition. In certain embodiments, the pharmaceutical composition comprises a mucoadhesive polymer or mucoadhesive excipient. Non-limiting examples of mucoadhesive polymers and mucoadhesive excipients include: hypromellose, lectin, thiolated polymers (e.g., chitosan-iminothiolane, poly(acrylic acid)-cysteine, poly(acrylic acid)-homocysteine, chitosan-thioglycolic acid, chitosan-thioethylamidine, alginate-cysteine, poly(methacrylic acid)-cysteine, and carboxymethylcellulose-sodium cysteine), polyethylene glycol, polyvinyl alcohol, polyvinylpyrrolidinone, polyacrylic acid (Carbopol™), polyhydroxyethyl methacrylate, chitosan, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl ... Examples of suitable mucoadhesive polymer excipients include cellulose, methylcellulose, sodium carboxymethylcellulose, aminated cornstarch, cellulose derivatives, poly(acrylic acid) polymers, poly(hydroxyethyl methylacrylate), poly(ethylene oxide), poly(vinylpyrrolidone), poly(vinyl alcohol), tragacanth, sodium alginate, karaya gum, guar gum, xanthan gum, soluble starch, gelatin, pectin, chitosan, methylcellulose, hyaluronic acid, hydroxypropyl methylcellulose, hydroxypropyl cellulose, gellan gum, carrageenan, cationic hydroxyethyl cellulose, hydrogel, dihydroxyphenylalanine, and alginate-polyethylene glycol acrylate. In certain embodiments, the pharmaceutical composition comprises about 0% to about 10% of a mucoadhesive polymer excipient selected from the list consisting of carbomer, polyethylene glycol, crospovidone, polycarbophil, hypromellose, and hydroxyethyl cellulose.

[0355] In certain embodiments, the pharmaceutical composition comprises at least about 0.1% to about 90%, about 92%, about 93%, about 95%, about 98%, about 97%, about 98%, or about 99% mucoadhesive polymer. In certain embodiments, the pharmaceutical composition comprises about 0.1% to about 1% mucoadhesive polymer. In certain embodiments, the pharmaceutical composition comprises about 0.5% to about 5% mucoadhesive polymer. In certain embodiments, the pharmaceutical composition comprises about 1% to about 10% mucoadhesive polymer. In certain embodiments, the pharmaceutical composition comprises about 5% to about 20% mucoadhesive polymer. In certain embodiments, the pharmaceutical composition comprises about 10% to about 50% mucoadhesive polymer. In certain embodiments, the pharmaceutical composition comprises about 20% to about 75% mucoadhesive polymer. In certain embodiments, the pharmaceutical composition comprises about 50% to about 90% mucoadhesive polymer. In certain embodiments, the pharmaceutical composition comprises about 75% to about 99% mucoadhesive polymer, hi certain embodiments, the pharmaceutical composition comprises at least about 0.1 percent, 0.25 percent, 0.5 percent, 0.75 percent, 1 percent, 2 percent, 3 percent, 5 percent, 10 percent, 15 percent, 20 percent, 25 percent, 30 percent, 35 percent, 40 percent, 45 percent, 50 percent, 55 percent, 60 percent, 65 percent, 70 percent, 75 percent, 80 percent, 85 percent, 90 percent, or 95 percent mucoadhesive polymer. In certain embodiments, the pharmaceutical composition comprises about 0.1 percent, 0.25 percent, 0.5 percent, 0.75 percent, 1 percent, 2 percent, 3 percent, 5 percent, 10 percent, 15 percent, 20 percent, 25 percent, 30 percent, 35 percent, 40 percent, 45 percent, 50 percent, 55 percent, 60 percent, 65 percent, 70 percent, 75 percent, 80 percent, 85 percent, 90 percent, or 95 percent or less of a mucoadhesive polymer. In certain embodiments, the pharmaceutical composition comprises 0% mucoadhesive polymer.In this case, adhesion to the target site is achieved through the use of other pharmaceutically acceptable excipients.

[0356] To prepare pharmaceutical compositions according to the present invention, a therapeutically effective amount of any active compound described herein, including, but not limited to, Compound I monofumarate, Compound II, or Compound III according to the present invention, is mixed with a pharmaceutically acceptable carrier to produce a dosage, often according to conventional pharmaceutical compounding techniques. The carrier can take a variety of forms, depending on the form of preparation desired for administration, e.g., topical, oral, or parenteral. When preparing pharmaceutical compositions for topical use, any of the usual pharmaceutical media can be used. Thus, for liquid or semi-solid topical preparations, such as gels, creams, ointments, suspensions, elixirs, and solutions, suitable carriers and additives, including water, glycols, oils, alcohols, preservatives, and the like, may be used. In certain embodiments, the pharmaceutical composition comprises propylene glycol. In certain embodiments, the pharmaceutical composition comprises carboxypolymethylene. In certain embodiments, the pharmaceutical composition comprises ethylenediaminetetraacetic acid (EDTA). In certain embodiments, the pharmaceutical composition comprises sorbic acid. In certain aspects, the pharmaceutical composition comprises carbomer. In certain embodiments, the pharmaceutical composition comprises hydroxyethylcellulose. In certain embodiments, the pharmaceutical composition comprises polyethylene glycol.

[0357] For solid external preparations such as powders, tablets, capsules, and suppositories, suitable carriers and additives may be used, including starches, sugar carriers (such as dextrose, mannitol, lactose, and related carriers), diluents, granulating agents, lubricants, binders, mucoadhesive polymers, disintegrants, and the like. If desired, tablets or capsules may be coated by standard techniques or may be sustained-release. The use of these dosage forms may significantly increase the bioavailability of the compound in patients. In certain embodiments, the pharmaceutical composition comprises mannitol. In certain embodiments, the pharmaceutical composition comprises magnesium stearate. In certain embodiments, the pharmaceutical composition comprises microcrystalline cellulose. In certain embodiments, the pharmaceutical composition comprises polycarbophil. In certain embodiments, the pharmaceutical composition comprises polyethylene oxide. In certain embodiments, the pharmaceutical composition comprises colloidal silicon dioxide. In certain embodiments, the pharmaceutical composition comprises povidone. In certain embodiments, the pharmaceutical composition comprises isopropyl alcohol. In certain embodiments, the pharmaceutical composition comprises sodium starch glycolate. In certain embodiments, the pharmaceutical composition comprises croscarmellose sodium. In certain embodiments, the pharmaceutical composition comprises crospovidone. In certain embodiments, the pharmaceutical composition comprises hydroxypropyl methylcellulose. In certain embodiments, the pharmaceutical composition comprises lactose. In certain embodiments, the powder pharmaceutical composition comprises one or more excipients selected from the group consisting of xanthan gum, microcrystalline cellulose, polyethylene oxide, hydroxypropyl methylcellulose, hydroxypropyl cellulose, sodium carboxymethylcellulose, povidone, mannitol, colloidal silicon dioxide, sodium benzoate, sodium starch glycolate, sodium lauryl sulfate, poloxamer 407, polyoxypropylene-polyoxyethylene copolymer, and the like.

[0358] In certain embodiments, a pharmaceutical composition comprising an effective amount of a fumarate salt of any active compound described herein, including but not limited to Compound I, may be formulated with an effective amount of acacia, agar, alginic acid, ascorbyl palmitate, bentonite, benzoic acid, butylated hydroxyanisole, butylated hydroxytoluene, butylene glycol, calcium acetate, calcium hydroxide, canola oil, carob bean gum, carrageenan, castor oil, cellulose, corn starch, edetate disodium, erythorbic acid, ethyl lactate, ethylcellulose, glycerin, glyceryl behenate, glyceryl monooleate, glyceryl monostearate, hydroxyethylmethylcellulose, hydroxypropylcellulose, hypromellose, lactic acid, lauric acid, lectin, linoleic acid, medium chain triglycerides, methylparaben, methylcellulose, microcrystalline cellulose, cellulose gum, cellulose gum, cellulose gum, cellulose gum ai ... The composition further comprises a pharmaceutically acceptable excipient selected from the list consisting of microcrystalline cellulose, microcrystalline wax, myristic acid, oleic acid, palmitic acid, peanut oil, pectin, phosphoric acid, polycarbophil, potassium alginate, propionic acid, propyl gallate, propylparaben, propylene glycol, propylene glycol alginate, silicon dioxide, simethicone, sodium alginate, sodium benzoate, sodium bicarbonate, sodium carboxymethylcellulose, sodium chloride, sodium citrate, sodium lactate, sodium lauryl sulfate, sodium metabisulfite, sodium phosphate, sodium sulfite, sodium thiosulfate, sorbic acid, stearic acid, talc, tapioca starch, tartaric acid, thymol, urea, vitamin E polysuccinate, beeswax, xanthan gum, and zinc acetate.

[0359] In certain embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable excipient for use as a pessary. In certain embodiments, a pharmaceutical composition comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprises up to 99.9% of a pessary excipient selected from the group consisting of hard fat, PEG, macrogol, cocoa butter, and glycerol. Non-limiting examples of hard fats include Ovucire™ (mono-, di-, and triglyceride esters of fatty acids (C 10 ~C 18 ), ethoxylated fatty alcohols with a predominant triester fraction), Witepsol™ (glycerol esters of saturated vegetable fatty acids such as lauric acid), and Supposi-base™ (a blend of saturated polyglycolylated glycerides).

[0360] In certain embodiments, a pharmaceutical composition comprising an effective amount of any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprises a pharmaceutically acceptable excipient that enhances the penetration, disintegration, film formation, and / or controlled release properties of the composition.

[0361] In certain embodiments, pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprise a penetration-enhancing excipient. In certain embodiments, the penetration-enhancing excipient is selected from the group consisting of oleic acid, eucalyptol, caprylol, Labrafil, Labrasol, lauroglycol, diethylene glycol monomethyl ether (Transcutol), propylene glycol, sodium laurate, sodium lauryl sulfate, cetyltrimethylammonium bromide, poloxamer (231, 182, 184), Tween 20, 40, 60, 80, fatty acids and fatty acid esters, isostearic acid, glycerin, and chitosan. In certain embodiments, a pharmaceutical composition comprising the fumarate salt of Compound I contains 0% to about 20% of a penetration-enhancing excipient selected from the group consisting of cetyl alcohol, propylene glycol, Transcutol P, oleic acid, isopropyl myristate, propylene glycol dicaprylate, glyceryl monooleate, propylene glycol monocaprylate, PEG-8 beeswax, cetyl alcohol, stearic acid, cetyl palmitate, and cetosteryl alcohol. In certain embodiments, the pharmaceutical composition contains about 0% to about 25% of a penetration-enhancing excipient selected from the list consisting of stearyl alcohol, polysorbate 80, sodium lauryl sulfate, mono- and diglycerides, sorbitan monostearate, glyceryl isostearate, polyoxyl 15 hydroxystearate, polyoxyl 40 hydrogenated castor oil, octyldodecanol, and soy lecithin.

[0362] In certain embodiments, a pharmaceutical composition comprising any of the active compounds described herein, including but not limited to, Compound I monofumarate, Compound II, or Compound III, further comprises a film-forming excipient. In certain embodiments, a pharmaceutical composition comprising any of the active compounds described herein, including but not limited to, Compound I monofumarate, Compound II, or Compound III, contains from 0% to about 99% of a film-forming excipient selected from the group consisting of hypromellose, polyethylene glycol, polymethacrylate, microcrystalline cellulose, guar gum, xanthan gum, and polyvinylpyrrolidone.

[0363] In certain embodiments, a pharmaceutical composition comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprises an excipient that allows for controlled release of the active compound. In certain embodiments, the controlled release pharmaceutical composition comprises ethyl cellulose, hypromellose, microcrystalline wax, polycarbophil, or beeswax.

[0364] Percentage ranges for excipients and other ingredients of pharmaceutical compositions are given as weight percent unless otherwise specified.

[0365] In certain embodiments, pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprise a disintegration-enhancing excipient. In certain embodiments, the disintegration-enhancing excipient is selected from the group consisting of cellulose, guar gum, crospovidone, polyplasdone, soy polysaccharide, calcium silicate, gelatin, cation exchange resin, bentonite, citrus pulp, alginic acid, calcium alginate, methylcellulose, microcrystalline cellulose, sodium carboxymethylcellulose, croscarmellose, solka floc, corn starch, sodium starch glycolate (Explotab, Primojel), glycine, hydroxypropyl starch, and starch 1500. In certain embodiments, the pharmaceutical composition comprises up to about 99% disintegration-enhancing excipient, such as mannitol and / or microcrystalline cellulose. In certain embodiments, the pharmaceutical composition comprises from about 0% to about 70% of a disintegration-enhancing excipient selected from the list consisting of lactose, sucrose, and calcium phosphate. In certain embodiments, the pharmaceutical composition comprises from about 0% to about 50% of a disintegration-enhancing excipient selected from the list consisting of sodium bicarbonate, citric acid, maleic acid, adipic acid, and fumaric acid. In certain embodiments, the pharmaceutical composition comprises from about 0% to about 20% of a disintegration-enhancing excipient selected from the list consisting of sodium starch glycolate, pregelatinized starch, crospovidone, and croscarmellose sodium.

[0366] In certain embodiments, pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprise 0% to about 70% mannitol, including any amount that achieves the desired result, including but not limited to, for example, up to about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, or about 70%. In certain embodiments, pharmaceutical compositions comprising the fumarate salt of Compound I further comprise 0% to about 70% lactose, including any amount that achieves the desired result, including but not limited to, for example, up to about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, or about 70%. In certain embodiments, pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprise 0% to about 70% sucrose, including any amount that achieves the desired result, including but not limited to, for example, up to about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, or about 70%. In certain embodiments, pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprise about 0% to about 70% microcrystalline cellulose, including any amount that achieves the desired result, including but not limited to, for example, up to about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, or about 70%. In certain embodiments, a pharmaceutical composition comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprises 0% to about 20% sodium starch glycolate, including but not limited to, any amount that achieves the desired result, for example, up to about 1%, about 2%, about 3%, about 5%, about 7%, about 10%, about 12%, about 15%, or about 20%.In certain embodiments, pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprise about 0% to about 20% pregelatinized starch, including but not limited to, any amount that achieves the desired result, for example, up to about 1%, about 2%, about 3%, about 5%, about 7%, about 10%, about 12%, about 15%, or about 20%. In certain embodiments, pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprise about 0% to about 20% crospovidone, including but not limited to, any amount that achieves the desired result, for example, up to about 1%, about 2%, about 3%, about 5%, about 7%, about 10%, about 12%, about 15%, or about 20%. In certain embodiments, pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprise about 0% to about 20% croscarmellose sodium, including but not limited to, any amount that achieves a desired result of, for example, up to about 1%, about 2%, about 3%, about 5%, about 7%, about 10%, about 12%, about 15%, or about 20%. In certain embodiments, pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to, Compound I monofumarate, Compound II, or Compound III, further comprise 0% to about 50% xanthan gum, including but not limited to, any amount that achieves a desired result of, for example, up to about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, or about 45%. In certain embodiments, a pharmaceutical composition comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprises 0% to about 50% polycarbophil, including but not limited to any amount that achieves the desired result, for example, up to about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, or about 45%.In certain embodiments, pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprise 0% to about 50% polyethylene oxide, including but not limited to, any amount that achieves a desired result of, for example, up to about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, or about 45%. In certain embodiments, pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprise 0% to about 50% hydroxyethyl methylcellulose, including but not limited to, any amount that achieves a desired result of, for example, up to about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, or about 45%. In certain embodiments, pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprise 0% to about 50% hydroxyethylcellulose, including but not limited to, any amount that achieves a desired result of, for example, up to about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, or about 45%. In certain embodiments, pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprise 0% to about 50% hypromellose, including but not limited to, any amount that achieves a desired result of, for example, up to about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, or about 45%. In certain embodiments, a pharmaceutical composition comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprises 0% to about 50% hydroxypropyl cellulose, including but not limited to any amount that achieves the desired result, for example, up to about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, or about 45%.In certain embodiments, pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprise 0% to about 50% PVP, including but not limited to, any amount that achieves a desired result of, for example, up to about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, or about 45%. In certain embodiments, pharmaceutical compositions comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprise 0% to about 50% microcrystalline cellulose, including but not limited to, any amount that achieves a desired result of, for example, up to about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, or about 45%.

[0367] In an exemplary embodiment according to the present invention, any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, is used to treat, prevent, or delay an HPV infection, or a secondary pathology, condition, or complication of HPV.

[0368] In certain embodiments, tablets used to treat, prevent, or delay HPV infection or secondary conditions, conditions, or complications containing any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further contain about 250 mg microcrystalline cellulose, about 20 mg crospovidone, about 5 mg magnesium stearate, about 5 mg silicon dioxide, about 5 mg polyethylene oxide, and about 100 mg mannitol. In certain embodiments, tablets used to treat, prevent, or delay HPV infection or secondary conditions, conditions, or complications of HPV, further contain about 155 mg microcrystalline cellulose, about 1.75 mg magnesium stearate, and about 17.5 mg mannitol.

[0369] In certain embodiments, a semi-solid formulation used to treat, prevent, or delay an HPV infection or a secondary pathology, condition, or complication of HPV comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprises about 15 mg of carbomer, about 50 mg of propylene glycol, about 10 mg of sorbic acid, about 5 mg of EDTA, and about 920 mg of water. In certain embodiments, a semi-solid formulation used to treat, prevent, or delay an HPV infection or a secondary pathology, condition, or complication of HPV comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprises about 20 mg of carbomer; about 70 mg of mineral oil; about 80 mg of a mixture of polyoxyl 6 stearate type I, ethylene glycol stearate, and polyoxyl 32 stearate type 1; about 5 mg of parabens; about 60 mg of propylene glycol; about 5 mg of EDTA; and about 760 mg of water.

[0370] In certain embodiments, a dry powder for reconstitution used to treat, prevent, or delay an HPV infection or a secondary pathology, condition, or complication of HPV comprising any of the active compounds described herein, including but not limited to Compound I monofumarate, Compound II, or Compound III, further comprises about 15.5 mg xanthan gum, about 19.8 mg mannitol, about 5 mg silicon dioxide, and about 0.5 mg sodium benzoate.

[0371] VII. Combination and Alternative Therapies The treatments described herein for intraepithelial neoplasia can be combined with traditional approaches, such as, but not limited to, excision or ablation of the transformed zone. Techniques include cryotherapy, laser therapy, loop electrosurgical procedure (LEEP), and cone biopsy. All of these surgical procedures can damage the affected area and cause scarring. LEEP, the most common intervention for cervical intraepithelial neoplasia, is effective in 60% to 90% of cases but is associated with a significantly increased risk of miscarriage, ectopic pregnancy, and negative psychological outcomes. In certain embodiments, the treatments described herein are used to reduce, ameliorate, or replace the use of these traditional practices.

[0372] In certain embodiments, the treatments described herein can be used in combination with surgical procedures. In certain embodiments, patients in need thereof can undergo surgery before, during, and / or after administration of an effective amount of a compound described herein. In certain embodiments, the surgical procedure can be excision of target and / or diseased tissue, including but not limited to loop electrosurgical excision procedure (LEEP), large loop excision of the transition zone (LLETZ), knife conization, cold knife conization, knife cone biopsy, or laser conization. In certain embodiments, the surgical procedure can be cauterization, including but not limited to laser cauterization or cryoablation.

[0373] The efficacy of a drug against HPV infection can be extended, increased, or restored by administering the compound in combination with or alternating with another (perhaps two or three other) antiviral compound that induces different mutations or acts through a pathway different from that of the principal drug. Alternatively, the pharmacokinetics, biodistribution, half-life, or other parameters of the drug can be altered by such combination therapy (which may include alternation therapy, if deemed coordinated). Furthermore, because HPV is associated with several types of cancer, combination therapy with anticancer therapeutics can provide better outcomes for patients. Because the disclosed compound II is a DNA polymerase inhibitor, it can be useful to administer the compound to a host in need thereof, for example, in combination with: a) protease inhibitors; b) another DNA polymerase inhibitor; c) inhibitors of E6 or E6AP, such as MEDI0457, luteolin, CAF-24, or gossypetin; d) inhibitors of E7; e) inhibitors of E1 or E2, including inhibitors of E1-E2 protein interactions; f) L2 lipopeptide; g) L1 or L2 inhibitors or degraders; h) HDAC inhibitors such as vorinostat; i) degrading agents for tetraspanins such as CD9, CD63 or CD151; j) Immunotherapy, such as T cell therapy (including adoptive T cell therapy) and checkpoint inhibitors; k) antiproliferative drugs; l) therapeutic vaccines; m) preventive vaccines; n) Trichloroacetic acid; o) salicylic acid; p) Imiquimod; q) Podofilox; r)Gardasil(trademark)9; s)Gardasil(trademark)4; t) Cervarix; u)VGX-3100; v) GGX-188E; and / or w)ADXS11-001. [Example]

[0374] Overview The following instrumental methods were used to characterize the morphic forms of the present invention.

[0375] TIFF2025526212000027.tif89170

[0376] TIFF2025526212000028.tif38170

[0377] TIFF2025526212000029.tif53170

[0378] TIFF2025526212000030.tif52170

[0379] Karl Fischer moisture determination Equipment:Mettler Toledo Coulometric KF Titrator C30 Method: coulometric measurement

[0380] TIFF2025526212000031.tif31170

[0381] TIFF2025526212000032.tif36170

[0382] TIFF2025526212000033.tif63170

[0383] TIFF2025526212000034.tif62170

[0384] TIFF2025526212000035.tif47170

[0385] TIFF2025526212000036.tif72170

[0386] TIFF2025526212000037.tif52170

[0387] Abbreviation: DMF: N,N-dimethylformamide; DCM: dichloromethane, methylene dichloride; MeOH: methanol, methyl alcohol; ACN: acetonitrile; EtOH: Ethanol; Ethyl alcohol; IPAc: isopropyl acetate; IPA: Isopropyl alcohol, isopropanol; THF: tetrahydrofuran; MEK: methyl ethyl ketone; DIAD: diisopropyl azodicarboxylate; DEAD: diethyl azodicarboxylate; MTBE: methyl tert-butyl ether; DMSO-d6: deuterated dimethyl sulfoxide; Cs2CO3: Cesium carbonate; TMSBr: trimethylsilyl bromide; NaOMe: sodium methoxide; TEA: triethylamine; Ph3P: triphenylphosphine; Na2SO4: sodium sulfate; NaOH: sodium hydroxide; HCl: Hydrochloric acid; H2SO4: sulfuric acid; BsOH: benzenesulfonic acid; p-TsOH: para-toluenesulfonic acid; MsOH: methanesulfonic acid; 1 1H NMR: proton nuclear magnetic resonance; LCMS: liquid chromatography mass spectrometry; HPLC: high pressure liquid chromatography.

[0388] 1H NMR sections: s = singlet, bs = broad singlet, d = doublet, dd = doublet of doublets, t = triplet, m = multiplet, J = spin-spin coupling constant

[0389] Example 1: Approximate solubility of the free base of Compound I at 25°C Approximately 2 mg of Compound I free form was weighed into a 2 mL glass vial, and 20 μL aliquots of each solvent (Table 1) were added to determine solubility at 25°C. In a second set of experiments, approximately 10 mg of Compound I free form was added to a 2 mL glass vial, and 20 μL aliquots of each solvent were added to determine solubility at 50°C. The maximum amount of each solvent added was 1 mL. Approximate solubility was determined visually. The results are shown in Table 1.

[0390] TIFF2025526212000038.tif81170

[0391] Example 2: Crystallization Screening for Compound I Free Base Form Equilibration with solvent at 25°C Approximately 30 mg of Compound I free base was equilibrated in a suitable amount of solvent as shown in Table 2 with stirring on a stir plate at 25° C. for 1 week. No precipitated solids were observed on day 8 of the experiment, and all 10 samples were stirred at 5° C. for approximately 3 days. In experiment 10, the suspension in heptane was filtered. The solid material (wet cake) obtained in experiment 10 was characterized by XRPD, DSC, TGA, and NMR.

[0392] TIFF2025526212000039.tif66170

[0393] According to XRPD measurements (Figure 1), the solid material obtained in Run 10 formed Compound I free form pattern 1. This pattern was characterized by DSC (Figure 2) as having an onset melting temperature of 40.3°C (with a transition enthalpy of 43 J / g) and by TGA (Figure 3) as having a 0.1% weight loss at 40°C. 1 It contained 0.2% residual heptane according to 1 H NMR.

[0394] Precipitation by addition of antisolvent Approximately 30 mg of the free base of Compound I was dissolved in the solvents listed below (Table 3). To the resulting solution, an anti-solvent was slowly added. The precipitate in Experiment 3 was collected by filtration and analyzed by XRPD.

[0395] TIFF2025526212000040.tif71170

[0396] According to the XRPD data (Figure 1), a solid material was formed as shown in Experiment 3, Table 3 Compound I free form pattern 1.

[0397] One crystalline free form, Compound I Free Form Pattern 1, was obtained from equilibration experiments in heptane and antisolvent experiments in acetone / MTBE.

[0398] Example 3: Synthesis of salts of Compound I Compound I sulfate Two methods were used to synthesize the sulfate salt of compound I.

[0399] Method A To a solution of compound I (0.049 g, 0.1 mmol) in dry THF (1 mL) at 0°C to 10°C, a solution of sulfuric acid (1 N / THF) was slowly added. During the addition of the sulfuric acid solution, the clear pink THF solution of compound I free base turned into an off-white semi-solid. The reaction mixture was allowed to warm to room temperature over 20 to 30 minutes and shaken. After the solid material settled, the supernatant was carefully decanted. The resulting semi-solid was washed with additional dry THF (2 × 2 mL), and the resulting solid was dried under high vacuum to give 0.053 g of compound I sulfate-1 as an off-white solid.

[0400] Method B To a solution of compound I (0.024 g, 0.05 mmol) in dry ethyl acetate (0.5 mL) at 0°C to 10°C, a solution of sulfuric acid (1N / EtOAc) was slowly added. During the addition of the sulfuric acid solution, the clear pink EtOAc solution of compound I free base turned into an off-white solid. The reaction mixture was allowed to warm to room temperature over 20 to 30 minutes and shaken vigorously. After the solid settled, the supernatant was carefully decanted. The off-white solid was washed with additional 2 x 2 mL of EtOAc. The resulting off-white solid was dried under high vacuum to give 0.023 g of compound I sulfate-2.

[0401] Compound I methanesulfonate Two methods were used to synthesize the methanesulfonate salt of compound I.

[0402] Method A To a solution of compound I (0.049 g, 0.1 mmol) in dry EtOAc (1 mL) at 0°C–10°C, neat methanesulfonic acid (MSA; MW = 96.11; d = 1.47; 0.007 mL; 0.11 mmol) was added dropwise. During the addition of the MSA solution, the pink, clear EtOAc solution of compound I free base turned into an off-white, semi-solid (glue-like). The heterogeneous mixture was allowed to warm to room temperature for 20–30 min and shaken vigorously. After allowing the semi-solid material to settle, the supernatant was carefully decanted. The resulting semi-solid (glue-like) was washed with methyl tert-butyl ether (MTBE; 2 × 2 mL) and dried under high vacuum to give 0.054 g of compound I methanesulfonate-1.

[0403] Method B To a solution of compound I (0.049 g, 0.1 mmol) in dry IPA (1 mL) at 0°C–10°C, methanesulfonic acid solution (1N in THF; 0.11 mmol; 0.110 mL) was added dropwise. During the addition of the MSA solution, the pink, clear EtOAc solution of compound I free base turned into an off-white solid. The heterogeneous mixture was allowed to warm to room temperature for 20–30 min and shaken vigorously. After allowing the solid material to settle, the supernatant was carefully decanted. The resulting solid was washed with methyl tert-butyl ether (MTBE; 2 × 2 mL) and dried under high vacuum to give 0.049 g of compound I methanesulfonate-2.

[0404] Compound I Hydrochloride To a solution of compound I (MW = 492; 0.049 g, 0.1 mmol) in dry EtOAc (1 mL) at 0°C to 10°C, HCl solution (4N in dioxane; 0.11 mmol; 0.027 mL) was added dropwise. During the addition of the HCl solution, the clear pink EtOAc solution of compound I free base turned into an off-white solid. This heterogeneous mixture was allowed to warm to room temperature for 20 to 30 minutes and shaken vigorously. After allowing the solid material to settle, the supernatant was carefully decanted. The resulting solid was washed with methyl tert-butyl ether (MTBE; 2 × 2 mL) and dried under high vacuum to give 0.045 g of compound I HCl salt.

[0405] Compound I monofumarate To a solution of compound I (0.035 g, 0.071 mmol) in dry isopropanol (0.1 mL) at 0°C to 10°C, fumaric acid (MW = 116; 12.3 mg; 0.106 mmol; 1.5 equiv.) was added. The reaction mixture was allowed to reach room temperature, then heated at 60°C for 30 min and stirred at room temperature for 12 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure at 40°C. The mixture was diluted with MTBE (2 mL), shaken well, and the MTBE was carefully decanted. The colorless solid was washed with additional MTBE (2 mL) and dried under high vacuum to give 0.022 g of compound I sesquifumarate. Compound I monofumarate can be synthesized by washing compound I sesquifumarate with additional MTBE and drying under high vacuum.

[0406] Compound I benzenesulfonate To a solution of compound I (0.057 g, 0.115 mmol) in dry EtOAc (1.0 mL) at 0°C to 10°C, a solution of benzenesulfonic acid (BsOH, MW = 158; 0.020 g; 0.127 mmol in 0.2 mL of EtOAc) was added dropwise. During the addition of the BsOH solution, the pink EtOAc solution of compound I free base precipitated as a colorless solid. The reaction mixture was allowed to warm to room temperature for 20 to 30 minutes and shaken vigorously. After the solid material settled, the supernatant was carefully decanted. The colorless solid was washed with additional MTBE (2 × 2 mL). The resulting solid was dried under high vacuum to give 0.064 g of compound I besylate salt.

[0407] Compound I tosylate To a solution of compound I (0.024 g, 0.05 mmol) in dry EtOAc (0.5 mL) at 0°C to 10°C, a solution of p-toluenesulfonic acid (p-TsOH, 0.055 mL; 0.055 mmol; 1N in EtOAc) was added dropwise. During the addition of the p-TsOH solution, the pink EtOAc solution of compound I free base precipitated as a colorless solid. The reaction mixture was allowed to warm to room temperature for 20 to 30 minutes and shaken vigorously. After the solid material settled, the supernatant was carefully decanted. The colorless solid was washed with additional MTBE (2 × 2 mL). The resulting solid was dried under high vacuum to give 0.027 g of compound I tosylate.

[0408] The melting points of the salts of Compound I obtained in this example were measured by differential scanning calorimetry, and the results are shown in Table 4. The monofumarate salt, produced by washing the sesquifumarate salt, has the highest melting point of the salts tested.

[0409] TIFF2025526212000041.tif51170

[0410] Example 5: Salt Screening of Compound I Free Base Eight acids (fumaric acid, citric acid, L-malic acid, L-tartaric acid, succinic acid, benzenesulfonic acid, oxalic acid, and maleic acid) and two coformers (L-proline and nicotinamide) were selected for screening of potential salt and / or cocrystal formation. Compound I was found to exist as two diastereomers ( 1 H NMR (purity: approximately 98-99%) was used as a mixture of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61

[0411] Salt screening by slurry equilibration Compound I free base as a mixture of diastereomers (approximately 30 mg) was added to a suitable solvent, followed by the addition of 1 molar equivalent or 0.5 molar equivalent (as indicated in Table 5 below) of the acid (Experiments RC2–RC10) or coformer (Experiments RC11 and RC12) with stirring at 50°C. The mixture was stirred at 50°C for 2 hours and then at 25°C for at least 12 hours. In experiments in which a clear solution formed, half of the solution was evaporated in a fume hood, and the remainder was treated by antisolvent addition. The resulting suspension was removed and centrifuged. The solids obtained in Experiments RC2 (Sample RC2-EA), RC3 (Sample RC3-EA), RC7 (Samples RC7-IPA and RC7-EA), and RC11 (Sample RC11-EA) were analyzed by XRPD. The results are shown in Table 5.

[0412] TIFF2025526212000042.tif110170

[0413] In experiment RC2, the solid material obtained with EA (sample RC2-EA) was shown by XRPD (Figures 7 and 10) to be a mixture of fumaric acid and a phase similar to hemifumarate Pattern 1 (sample AS3-B in Table 6). In experiment RC3, the solid material obtained with EA (sample RC3-EA) was shown by XRPD data to be a phase similar to hemifumarate Pattern 1 (Figure 7). In experiment RC11, the solid material obtained with EA was shown to be a mixture of L-proline coformers and free form Pattern 1. In experiment RC7, the solid material obtained from isopropanol (sample RC7-IPA, Figure 8) and ethyl acetate (sample RC7-EA, Figure 9) was characterized by XRPD as a phase similar to monosuccinate Pattern 1 (see Table 6, experiment AS7, sample AS7-B, below).

[0414] Salt screening results by adding antisolvent An anti-solvent (methyl tert-butyl ether (MTBE) or heptane) was slowly added to the clear solutions obtained in the experiments in Table 5 (Experiments AS1-AS12 in Table 6). For Experiments AS2, AS3, and AS7, the addition of the heptane anti-solvent to the solutions in EtOH resulted in the precipitation of solid materials AS2-B, AS3-B, and AS7-B, which were characterized by XRPD.

[0415] Next, to clear solutions (from experiments AS1, AS3, and AS12) and sample AS5-A, which contained a small amount of solid material obtained after adding the antisolvent, 0.5 molar, 1 molar, or 1.5 molar amounts of acid or coformer were added with stirring at 25°C, and the mixtures were stirred at this temperature for at least 12 hours. No solid phase formed in experiments AS13-AS17. The results are summarized in Table 6.

[0416] TIFF2025526212000043.tif164170

[0417] Addition of heptane anti-solvent to the clear solution obtained from experiment RC2 (ethanol as solvent, 1 molar equivalent of fumaric acid) resulted in the formation of monofumarate pattern 1, characterized by XRPD (sample AS2-B, Figure 10). The composition of this material as the monofumarate was 1 The H NMR (ratio of free base form to fumaric acid = 1:0.98) established that no free fumaric acid phase was found in this sample by XRPD. Addition of heptane antisolvent to the clear solution obtained from experiment RC3 (ethanol as solvent, 0.5 molar equivalents of fumaric acid) resulted in the formation of hemifumarate pattern 1 (sample AS3-B, Figure 10), characterized by XRPD. The composition of this material as a hemifumarate was 1 The H NMR (ratio of free base form to fumaric acid = 1:0.54) established that no free fumaric acid phase was found in this sample by XRPD. Addition of heptane antisolvent to the clear solution obtained from entry RC7 (ethanol as solvent, 1 molar equivalent of succinic acid) resulted in the formation of monosuccinate pattern 1 (sample AS7-B, Figure 11) characterized by XRPD. The composition of this material as a monosuccinate was 1 This was established by 1 H NMR (ratio of free base form to succinic acid = 1:1.06) and by XRPD no free succinic acid phase was found in this sample.

[0418] Re-equilibration experiments The solvent was evaporated from the gel-like or oily samples obtained in the above salt screening by adding an antisolvent. The remaining mixture was slurried in MTBE or heptane at 25°C as shown in Table 7. The experimental results are shown in Table 7.

[0419] TIFF2025526212000044.tif66170

[0420] In re-equilibration experiments, citric acid, L-malic acid, L-tartaric acid, oxalic acid, maleic acid, and nicotinamide did not form crystalline phases with Compound I.

[0421] Example 6: Compound I Salt Production by Slurry Crystallization In experiment RC13, approximately 50 mg of Compound I free base was added to EtOH, and 0.5 molar fumaric acid was added with stirring at 50°C for 2 hours, followed by stirring at 25°C for at least 12 hours. Seeds from sample AS3-B were added, and heptane (0.4 mL) was added as an antisolvent. The resulting mixture was stirred at 5°C for approximately 3 days. The suspension was then removed and centrifuged. The resulting solid was dried in an oven under vacuum at 50°C for approximately 1 hour and analyzed by XRPD (Figure 12), NMR, DSC (Figure 13), and TGA (Figure 14).

[0422] In experiment RC14, approximately 50 mg of the free base of Compound I was added to EtOH, and an equimolar amount of succinic acid was added with stirring at 50°C for 2 hours, followed by stirring at 25°C for at least 12 hours. Seeds from sample AS7-B were added, and heptane (0.2 mL) was added as an antisolvent. The resulting suspension was removed and centrifuged. The resulting solid was dried in an oven under vacuum at 50°C for approximately 1 hour and analyzed by XRPD (Figure 15), NMR, DSC (Figure 16), and TGA (Figure 17).

[0423] In experiment RC15, approximately 50 mg of Compound I free base was added to 0.1 mL of EtOH. An equimolar amount of benzenesulfonic acid was dissolved in 0.2 mL of EtOH. The solution of benzenesulfonic acid was then added dropwise to the free base solution at 25 °C. MTBE (0.8 mL) was added as an antisolvent, and the mixture was stirred at 5 °C, resulting in the formation of some solids.

[0424] In experiment RC16, approximately 50 mg of Compound I free base was added to 0.1 mL of IPA. 1.5 molar amounts of fumaric acid were added to the mixture, and the mixture was stirred at 25°C. After approximately 3 hours, the sample became too sticky, so an additional 0.1 mL of IPA was added, and stirring was maintained at 25°C for approximately 22 hours. The sticky sample was obtained, dried in a vacuum oven at 50°C for approximately 2 hours, and reslurried in 1.0 mL of MTBE at 25°C for approximately 5 days. The resulting suspension was removed and centrifuged. The resulting solid was dried in an oven under vacuum at 40°C for approximately 1 hour and analyzed by XRPD (Figure 18) and NMR. The residual solid was added as a seed in RC18.

[0425] In experiment RC17, approximately 30 mg of Compound I free base was added to 0.1 mL of EtOH. 0.5 molar amount of succinic acid was added to the mixture while stirring at 50°C for 2 hours, resulting in a clear solution. Heptane (0.3 mL) was added as an anti-solvent. An oil was obtained, and the mixture was stirred at 25°C for approximately 5 days. The resulting suspension was removed and centrifuged. The resulting solid was dried in an oven under vacuum at 40°C for approximately 1 hour and analyzed by XRPD (Figure 19), NMR, DSC (Figure 20), and TGA (Figure 21).

[0426] In experiment RC18, approximately 30 mg of Compound I free base was added to 0.1 mL of IPA. An equimolar amount of fumaric acid was added to the mixture while stirring at 50°C for 2 hours, resulting in a clear yellow solution, and 0.3 mL of heptane was added as an antisolvent. Some seeds of RC16 were added. A suspension was obtained, which was stirred at 25°C for at least 12 hours. The resulting suspension was removed, centrifuged, and analyzed by XRPD (Figure 22). Monofumarate pattern 1 was obtained and added as seeds to the scale-up sample.

[0427] TIFF2025526212000045.tif160170

[0428] From the salt screening experiments, four patterns were identified: hemifumarate pattern 1, monofumarate pattern 1, hemisuccinate pattern 1, and monosuccinate pattern 1. The four patterns exhibit similar properties, such as low to moderate crystallinity and relatively low melting points.

[0429] Example 7: Preparation of fumarate salt Preparation of Compound I Hemifumarate Pattern 1 Approximately 200 mg of Compound I free base was added to 0.5 mL of EtOH. While stirring, 0.5 molar amounts of fumaric acid were added at 50° C., and the mixture was stirred for 2 hours. A clear solution was obtained. The solution was then cooled to 25° C. within 1 hour. Hemi-fumarate seeds from sample RC13 (Example 5, Table 7) were added, followed by the addition of 2.5 mL of heptane to induce precipitation. An oil was obtained and stirred at 25° C. for approximately 4 days. After 4 days, the resulting suspension was cooled to 5° C. After stirring at 5° C. for approximately 4 days, the precipitated solid was collected by filtration and dried under vacuum at 40° C. for approximately 3 hours. As a result, 116 mg of pale orange hemifumarate pattern 1 was obtained in 52% yield. The XRPD is shown in FIG. 23, the DSC is shown in FIG. 24, and the TGA is shown in FIG. 25.

[0430] Preparation of Compound I Monofumarate Pattern 1 (Small-Scale Preparation) Approximately 244 mg of Compound I free base was added to 0.8 mL of IPA. Then, 1.0 equivalent of fumaric acid was added with stirring at 50° C. for approximately 1.5 hours. The resulting clear yellow solution was cooled to 25° C. and stirred for approximately 5 minutes. Monofumarate seeds from sample RC18 (Table 7) were added to the mixture, followed by 4 mL of heptane as an antisolvent. The mixture was stirred at 25° C. for 4 days. The precipitated material was collected by filtration and dried under vacuum at 40° C. for approximately 2 hours. As a result, 208 mg of monofumarate Pattern 1 solid was obtained in 69% yield. The XRPD is shown in Figure 23. The DSC at 10° C. / min is shown in Figure 26. The DSC at 2° C. / min is shown in Figure 27b, the DSC cycle results are shown in Figure 27c, and the TGA is shown in Figure 28.

[0431] TIFF2025526212000046.tif95170

[0432] Compound I Pattern 1 is highly crystalline. The hemifumarate and monofumarate patterns are moderately crystalline.

[0433] Compound I Pattern 1 is an anhydride, T onset The compound I has a melting peak at 75.0°C with an enthalpy of about 64 J / g. The compound I pattern 1 shows a weight loss of about 0.3% at about 70°C. The KF shows that it contains about 1.7% water. The compound I contains about 0.7% (by weight) MTBE residue. 1 1 H NMR.

[0434] Compound I hemifumarate pattern 1 is an anhydride. Stoichiometry of the free form: fumaric acid is 1 Based on the results of H NMR, the ratio is approximately 1:0.5. onset It has a melting peak at 85.2°C with an enthalpy of about 37 J / g. It shows a weight loss of about 1.0% at about 85°C. KF shows that it contains about 1.7% water. About 0.7% of EtOH and 0.7% of heptane (by residual weight) 1 1 H NMR.

[0435] Compound I monofumarate pattern 1 is an anhydride. Stoichiometry of the free form: fumaric acid is 1 Based on the H NMR results, the ratio is approximately 1:1.0. onset The melting peak is at 107.2°C with an enthalpy of about 78 J / g. The two thermal events cannot be resolved even at heating rates of 2 K / min and 0.5 K / min by DSC. The weight loss is about 0.3% at about 107°C. KF indicates that the solution contains about 1.2% water. The residue is about 0.7% IPA and 2.2% heptane (by weight). 1 1 H NMR.

[0436] Example 8: Stability of Compound I Free Base Pattern 1, Compound I Monofumarate Pattern 1, and Compound I Hemifumarate Pattern 1 Initial Chemical Purity: The initial purities of Compound I free base pattern 1, Compound I monofumarate pattern 1 and Compound I hemifumarate pattern 1 are 98.7%, 98.6% and 97.8%, respectively.

[0437] Bulk stability: Accelerated stability studies were conducted in open containers at 25°C / 92% RH and 40°C / 75% RH, and in airtight containers at 60°C for 1 week. The results are shown in Table 10. The results for Compound I monofumarate Pattern 1 are also shown in Figure 28.

[0438] All three candidates showed good physical stability after exposure to the three conditions. All showed some decomposition after one week of exposure to 40°C / 75% RH, and two were not stable at higher temperatures (60°C for one week). Compound I hemifumarate pattern 1 tended to decompose more than Compound I monofumarate pattern 1 under the above two conditions.

[0439] TIFF2025526212000047.tif85170

[0440] Example 9: Solubility of Compound I Free Base Pattern 1, Compound I Monofumarate Pattern 1, and Compound I Hemifumarate Pattern 1 Approximately 4 mg of Compound I Pattern 1 was added to 2 mL of buffer solution. For Compound I hemifumarate Pattern 1 and Compound I monofumarate Pattern 1, approximately 4 mg was added to 1.8 mL and 1.6 mL of buffer solution, respectively. The pH value was adjusted to simulated vaginal fluid using 0.2 N NaOH. After stirring at 37°C for 0.5 and 2 hours, clear solutions were obtained for all three candidates.

[0441] TIFF2025526212000048.tif94170

[0442] Solubility was tested in five media: pH 3.0 citrate buffer, pH 4.5 acetate buffer, pH 6.8 phosphate buffer, water, and simulated vaginal fluid (pH 4.2) for 0.5 and 2 hours at 37° C. All three candidates were highly soluble in the media (>2 mg / mL).

[0443] Example 10: Hygroscopicity of Compound I Free Base Pattern 1, Compound I Hemifumarate Pattern 1, and Compound I Monofumarate Pattern 1 Hygroscopicity was investigated by DVS at 25°C using the following method. Method: 40-0-95-0-40%RH, dm / dt=0.002

[0444] The results are shown in Table 12 and Figures 29 to 34.

[0445] TIFF2025526212000049.tif123170

[0446] The moisture absorption was examined by DVS at 25°C. All three patterns are moderately hygroscopic. Compound I Pattern 1 shows approximately 4.3% water absorption up to 90% RH, but absorbs approximately 14.3% water at 95% RH. Compound I monofumarate Pattern 1 and Compound I hemifumarate Pattern 1 show approximately 2.9% and 6.9% water absorption up to 95% RH, respectively. There is no change in morphology after the DVS test.

[0447] Example 11: Large-scale preparation of monofumarate pattern 1 Approximately 4.16 g of Compound I free base was dissolved in 11 mL of IPA. 1.0 equivalent of fumaric acid was then added to the clear yellow solution while stirring at 50°C. After approximately 1 hour, some solid precipitated. 10 mg of monofumarate seeds (from the "Small-Scale Preparation" section above) were then added. The mixture was stirred at 50°C for approximately 1.5 hours, cooled to 25°C, and then stirred at 25°C for approximately 10 minutes. 40 mL of heptane was then added as an anti-solvent. The resulting suspension was stirred at 25°C for approximately 24 hours, then cooled to 5°C at a rate of 0.1°C / min, and stirred at 5°C for approximately 1 day. The solid was collected by filtration and dried in an oven under vacuum at 40°C for approximately 2 hours. Approximately 4.1 g of a pale pink solid was obtained in 81.2% yield. The XRPD is shown in Figure 35, the DSC at a rate of 10°C / min is shown in Figure 36a, the DSC at a rate of 2°C / min is shown in Figure 36b, the mDSC thermogram is shown in Figure 36c, and the TGA is shown in Figure 37.

[0448] TIFF2025526212000050.tif112170

[0449] Example 12: Polymorph Screening Study of Compound I Fumarate A polymorph screening study was conducted on the fumarate salt of Compound I (a mixture of diastereomers). Their polymorphic behavior was investigated by equilibration, precipitation by addition of antisolvent, slow cooling, fast cooling, and slow evaporation experiments.

[0450] Approximate solubility of Compound I monofumarate pattern 1 at 25°C and 50°C Approximately 2 mg of Compound I monofumarate pattern 1 (Example 11) was weighed into a 2 mL glass vial, and a 20 μL aliquot of each solvent (shown in Table 14) was added to determine the solubility at 25° C. Approximately 10 mg of Compound I monofumarate pattern 1 (Example 11) was weighed into a 2 mL glass vial, and a 20 μL aliquot of each solvent (shown in Table 14 below) was added to determine the solubility at 50° C. The maximum amount of each solvent added was 1 mL. The approximate solubility was determined visually.

[0451] TIFF2025526212000051.tif85170

[0452] Equilibration with solvent for 2 and 3 weeks at 25°C Approximately 30 mg of Compound I monofumarate Pattern 1 (obtained in Example 11) was equilibrated in an appropriate amount of solvent using a stir plate at 25°C for 2 and 3 weeks. The resulting suspension was filtered. The solid portion (wet cake) was examined by XRPD. If differences were observed, additional testing (e.g., NMR, DSC, TGA, HPLC, and PLM) was performed. The results are shown in Tables 15 to 30 and Figures 38 to 60.

[0453] TIFF2025526212000052.tif69170

[0454] For pattern D, the XRPD diffractogram is shown in Figures 38 and 39, the DSC thermogram is shown in Figures 40 and 41, and the TGA thermogram is shown in Figures 42 and 43.

[0455] TIFF2025526212000053.tif78170

[0456] For pattern C, the XRPD diffractogram is shown in FIG. 44, the DSC thermogram is shown in FIG. 45, and the TGA thermogram is shown in FIG.

[0457] TIFF2025526212000054.tif78170

[0458] For pattern B, the XRPD diffractogram is shown in FIG. 47, the DSC thermogram is shown in FIG. 48, and the TGA thermogram is shown in FIG.

[0459] TIFF2025526212000055.tif31170

[0460] The XRPD diffractogram is shown in Figure 47 (3 weeks).

[0461] TIFF2025526212000056.tif88170

[0462] For pattern 1, the XRPD diffractogram is shown in FIG. 50, the DSC thermogram is shown in FIG. 51, and the TGA thermogram is shown in FIG.

[0463] TIFF2025526212000057.tif94170

[0464] For pattern E, the XRPD diffractogram is shown in Figure 53, the DSC thermogram is shown in Figures 54 and 55, and the TGA thermogram is shown in Figures 56 and 57.

[0465] TIFF2025526212000058.tif60170

[0466] The XRPD diffractogram is shown in Figure 47 (3 weeks).

[0467] TIFF2025526212000059.tif89170

[0468] The XRPD diffractogram is shown in FIG.

[0469] TIFF2025526212000060.tif79170

[0470] The XRPD diffractogram is shown in FIG.

[0471] TIFF2025526212000061.tif99170

[0472] The XRPD diffractogram is shown in FIG.

[0473] TIFF2025526212000062.tif60170

[0474] The XRPDs are shown in Figures 58 and 59.

[0475] TIFF2025526212000063.tif92170

[0476] The XRPD diffractogram is shown in FIG.

[0477] TIFF2025526212000064.tif59170

[0478] The XRPDs are shown in Figures 58 and 59.

[0479] TIFF2025526212000065.tif71170

[0480] The XRPDs are shown in Figures 58 and 59.

[0481] TIFF2025526212000066.tif55170

[0482] The XRPD diffractogram is shown in FIG.

[0483] TIFF2025526212000067.tif97170

[0484] The XRPD diffractogram is shown in FIG.

[0485] Precipitation by addition of antisolvent Approximately 30 mg of Compound I monofumarate pattern 1 (Example 7) was dissolved in a good solvent. A poor solvent was slowly added to the resulting solution. The precipitate was collected by filtration. The solid portion (wet cake) was examined by XRPD. If differences were observed, further investigations were carried out (e.g., NMR, DSC, TGA). If no precipitate was obtained, the solution was cooled to 5°C for crystallization. After stirring at 5°C for approximately 23 days, no precipitate was obtained, and the solution was placed in a -20°C freezer for crystallization.

[0486] The results are shown in Table 31. The XRPD diffractograms are shown in Figures 61 and 62.

[0487] TIFF2025526212000068.tif163170

[0488] Crystallization at room temperature by slow evaporation In conjunction with the approximate solubility experiments, the solubility samples were filtered through a 0.45 μm nylon filter. The resulting solutions were allowed to slowly evaporate at ambient conditions. The polymorphic forms of the solid residues were investigated.

[0489] The results are shown in Table 32. The XRPD diffractograms are shown in Figures 63 and 64.

[0490] TIFF2025526212000069.tif71170

[0491] Crystallization from high-temperature saturated solutions by slow cooling Approximately 30 mg of Compound I monofumarate pattern 1 (Example 7) was dissolved in a minimum amount of selected solvent at 50°C. The resulting solution was cooled to 5°C at a rate of 0.1°C / min, and the precipitate was collected by filtration. The solid portion (wet cake) was examined by XRPD. If differences were observed, additional investigations were performed (e.g., NMR, DSC, TGA). If no precipitate was obtained, the solution was placed in a -20°C freezer for crystallization. After storing in a -20°C freezer for approximately 5 days, no precipitate was obtained, and heptane was added as an anti-solvent. The precipitate was collected by filtration. The solid portion (wet cake) was examined by XRPD.

[0492] The results are shown in Table 33. The XRPD diffractograms are shown in Figures 65 and 66.

[0493] TIFF2025526212000070.tif147170

[0494] Crystallization from high-temperature saturated solutions by rapid cooling Approximately 30 mg of Compound I monofumarate pattern 1 (Example 7) was dissolved in the minimum amount of selected solvent at 50°C. The resulting solution was placed in an ice bath and stirred. The precipitate was collected by filtration. The solid portion (wet cake) was examined by XRPD. If differences were observed, additional investigations were performed (e.g., NMR, DSC, TGA). If no precipitate was obtained, the solution was placed in a -20°C freezer for crystallization. After storing in a -20°C freezer for approximately 7 days, no precipitate was obtained, and heptane was added as an anti-solvent. The precipitate was collected by filtration. The solid portion (wet cake) was examined by XRPD.

[0495] The results are shown in Table 34. The XRPD diffractograms are shown in Figures 67 and 68.

[0496] TIFF2025526212000071.tif84170

[0497] Behavior of Compound I Monofumarate Pattern 1 under Heating and Cooling. The polymorphic behavior of compound I monofumarate pattern was investigated by two different heating-cooling cycles of DSC.

[0498] Cycle 1: 0°C to 106°C at 10°C / min; 106°C to 0°C at 10°C / min; reheat from 0°C to 250°C at 10°C / min.

[0499] Cycle 2: 0°C to 130°C at 10°C / min; 130°C to 0°C at 10°C / min; reheat from 0°C to 250°C at 10°C / min.

[0500] The results are shown in Table 35 and Figures 69 and 70.

[0501] TIFF2025526212000072.tif88170

[0502] Behavior under compression Approximately 10 mg of Compound I monofumarate pattern 1 (Example 7) was compressed in a hydraulic press at 10 MPa for 5 minutes. XRPD characterization was performed to determine polymorphic behavior under compression. No morphological changes were observed by XRPD.

[0503] Grinding simulation experiment Approximately 10 mg of Compound I monofumarate pattern 1 (Example 7) was manually ground with a mortar and pestle for 5 minutes. The morphology change and crystallinity were evaluated by XRPD. According to XRPD, no morphology change was observed, and the crystallinity was slightly decreased.

[0504] Granulation simulation experiment Water or ethanol was added dropwise to approximately 10 mg of Compound I monofumarate Pattern 1 (Example 7) until the solid was sufficiently wetted. The sample was manually ground using a mortar and pestle for 3 minutes. The sample was dried under ambient conditions for 10 minutes, and the morphology and crystallinity were evaluated by XRPD. No changes were observed in either ethanol or water by XRPD.

[0505] Summary of identified monofumarate and hemifumarate polymorphs Compound I monofumarate pattern 1 used in this study was prepared from Compound I free base according to Example 7. The initial form of monofumarate used in the polymorphism study (pattern 1) described below is the monofumarate anhydride with an HPLC purity of approximately 99.3%. The ratio of the free form to fumaric acid is 1 The ratio is about 1:0.96 by H NMR. Differential scanning calorimetry (DSC) shows a T of about 98.5°C with an enthalpy of about 14 J / g and a T of about 109.6°C with an enthalpy of about 25 J / g. onset Thermogravimetric analysis (TGA) showed that pattern 1 exhibited a weight loss of about 0.5% at about 98°C and a weight loss of 0.6% between 98°C and 140°C. Residues of about 1.0% (by weight) heptane and 0.2% (by weight) IPA were observed. 1 Karl Fischer water content as determined by HNMR indicates that it contains approximately 1.3% water.

[0506] During the polymorphism study, four new patterns were identified. Although the monofumarate was used as the initial physical form, the new patterns obtained showed different stoichiometries. Patterns B, C, and E are the hemifumarate salt, and Pattern D is a degradation product.

[0507] Pattern B is the anhydrous form of hemifumarate with an HPLC purity of about 99.6%. It can be obtained from equilibration experiments, anti-solvent addition, slow cooling, and fast cooling experiments using MEK, acetone, acetone / heptane, MEK / heptane, and EtOH / MTBE as solvents. The ratio of the free base form to fumaric acid is about 1:0.52 by 1H-NMR. The T is about 77.4°C with an enthalpy of about 71 J / g, and 88.4°C with an enthalpy of about 18 J / g. onset The sample has two thermal events: a weight loss of about 0.7% at about 77°C and a weight loss of 4.2% between 77°C and 130°C. Approximately 4.6% (by weight) of MEK residue remains. 1 1 H NMR.

[0508] A mixture of Pattern C and fumaric acid was obtained by equilibration, slow cooling, and fast cooling experiments using ACN and ACN / water as solvents. The ratio of free base form to fumaric acid in this mixture was 1 H NMR gives a ratio of about 1:0.95. After washing with water, the ratio decreased to about 1:0.76, indicating that Pattern C is not a monofumarate. The T onset , and a T of about 90.6°C with an enthalpy of about 15 J / g onset The sample has two thermal events: a weight loss of about 0.4% at about 73°C and a weight loss of 2.1% between 73°C and 144°C. ACN residue of about 2.0% (by weight) is observed. 1 1 H NMR.

[0509] Pattern D is a degradation product with an HPLC purity of about 0.2%. It was obtained in water by equilibration experiments. Its T is about 41.4°C with an enthalpy of about 67 J / g. onset , and a T of about 72.1°C with an enthalpy of about 29 J / g onset There are two thermal events: a weight loss of about 0.6% at about 41°C and a weight loss of 8.5% between 41°C and 178°C.

[0510] Pattern E is the anhydrous hemifumarate with an HPLC purity of approximately 98.9%. It was obtained by equilibration experiments in acetone / toluene. The ratio of the free base form to fumaric acid in this mixture is 1 H NMR gives a ratio of about 1:0.69. The T is about 53.1°C with an enthalpy of about 33 J / g. onset , and a T of about 96.5°C with an enthalpy of about 34 J / g onset It has two thermal events: a weight loss of about 1.0% at about 53°C and a weight loss of 3.6% between 53°C and 96°C. There is an acetone residue of about 0.6% (by weight). 1 1 H NMR.

[0511] The results of testing for the monofumarate and hemifumarate polymorphs of Compound I are summarized in Table 36 below. The salt ratio is the ratio of Compound I free base to salt counterion. "AS" indicates that the form can be prepared by antisolvent addition using the solvent / antisolvent pair listed in the table. "EQ" indicates that the form can be prepared by equilibration in the listed solvent. "SC" indicates that the form can be prepared by slowly cooling a solution of Compound I monofumarate in the listed solvent. "FC" indicates that the form can be prepared by rapidly cooling a solution of Compound I monofumarate in the listed solvent.

[0512] TIFF2025526212000073.tif125170

[0513] Example 13: Preparation of Compound II Pattern 1 Experiment 1. Small-scale synthesis and seed preparation 100mg of R P Compound I free base and 0.3 mL of IPA were added to a glass vial. To this was added 1.0 equivalent of fumaric acid, and the resulting mixture was stirred at 50°C for 2 minutes to precipitate most of the material. To this was added 1.0 mL of heptane. The sample was stirred at 50°C for 1 hour and then cooled to 3°C at 0.1°C / min. After stirring at 3°C for approximately 8 hours, 0.4 mL of heptane was added. The resulting solid was isolated by filtration and dried in a vacuum oven at 40°C for approximately 2 hours to obtain Compound II Pattern 1. Characterization results are reported in Table 37. The XRPD diffractogram of Compound II Pattern 1 is shown in Figure 71.

[0514] TIFF2025526212000074.tif41170

[0515] Experiment 2. Large-scale preparation 3g R PCompound I free base and 9 mL of IPA were added to a glass vial. 1.0 equivalent of fumaric acid was added, and the resulting mixture was stirred at 50°C for 5 minutes. Approximately 21 mg of Compound II Pattern 1 seeds from Experiment 1 were added to the mixture. 30 mL of heptane was added to the mixture. The sample was stirred at 50°C for 1 hour and then cooled to 3°C at 0.1°C / min. After stirring at 3°C for approximately 16 hours, the resulting solid was isolated by filtration and dried in a vacuum oven at 40°C for approximately 4 hours and at 50°C for approximately 3 hours. This afforded 2.9 g of Compound II Pattern 1 as a white solid in 78.3% yield. The XRPD diffractogram of Compound II Pattern 1 is shown in Figure 71. The DSC thermogram of Compound II Pattern 1 is shown in Figure 72. The TGA thermogram of Compound II Pattern 1 is shown in Figure 73.

[0516] TIFF2025526212000075.tif55170

[0517] Compound II Pattern 1 is an anhydride. The stoichiometry of the free base:fumaric acid is 1 Based on the results of H NMR, the ratio is approximately 1:1.0. onset The melting peak at 141.5°C is associated with decomposition. TGA shows a weight loss of about 0.3% at about 130°C. Residual solvent is 1 Not detected by 1 H NMR.

[0518] Example 14: Preparation of Compound IV Pattern 1 66mg of R P Compound I free base and 0.2 mL of EtOH were added to a glass vial. 0.5 equivalents of fumaric acid were added, and the resulting mixture was stirred at 50° C. for 2 hours, after which some solid was allowed to precipitate. 0.4 mL of heptane was added to the suspension, and the mixture was further stirred at 25° C. for approximately 4 days. The solid was isolated by filtration and dried in a vacuum oven at 50° C. for approximately 2 hours. The resulting Compound IV Pattern 1 was characterized as reported in Table 39. The XRPD diffractogram of Compound IV Pattern 1 is shown in Figure 74. The DSC thermogram of Compound IV Pattern 1 is shown in Figure 75. The TGA thermogram of Compound IV Pattern 1 is shown in Figure 76.

[0519] TIFF2025526212000076.tif50170

[0520] Example 15: Preparation of Compound III Pattern 1 S P 100 mg of compound I free base and 0.3 mL of IPA were added to a glass vial. 1.0 equivalent of fumaric acid was added, and the mixture was stirred at 50°C for 15 minutes to precipitate most of the material. After adding 1.0 mL of heptane, the sample was stirred at 50°C for 1 hour and then cooled to 3°C at 0.1°C / min. After stirring at 3°C for approximately 8 hours, 0.4 mL of heptane was added to the mixture to obtain a good suspension. The white solid was isolated by filtration and dried in a vacuum oven at 40°C for approximately 2 hours to obtain compound III Pattern 1. Characterization results are reported in Table 40. The XRPD diffractogram of compound III Pattern 1 is shown in Figure 77. The DSC thermogram of compound III Pattern 1 is shown in Figure 78. The TGA thermogram of compound III Pattern 1 is shown in Figure 79.

[0521] TIFF2025526212000077.tif40170

[0522] Example 16: Preparation of Compound III Pattern 2 3g of S PCompound I free base and 9 mL of IPA were added to a glass vial. After adding 1.0 equivalent of fumaric acid, a large amount of solid precipitated immediately. 30 mL of heptane was added to the mixture, followed by approximately 20 mg of compound III Pattern 1 seeds. The sample was stirred at 50°C for 1 hour and then cooled to 3°C at 0.1°C / min. After stirring at 3°C for approximately 20 hours, the sample was heated from 3°C to 50°C within 20 minutes, and then 0.2 equivalents of fumaric acid and 1.5 mL of heptane were added to the mixture. The resulting mixture was stirred at 50°C for approximately 2 hours and then cooled to 3°C at 0.1°C / min. After stirring at 3°C for approximately 13 hours, it was reheated to 50°C within 20 minutes and stirred at 50°C for approximately 6 hours. It was cooled to 3°C at 0.1°C / min and stirred at 3°C for approximately 2 days. The resulting solid was isolated by filtration and dried in a vacuum oven at 50° C. for approximately 3 hours to give 3.1 g of a white solid in 83.6% yield. The results are reported in Table 41. The XRPD diffractogram of Compound III Pattern 2 is shown in Figure 80. The DSC thermogram of Compound III Pattern 2 is shown in Figure 81. The TGA thermogram of Compound III Pattern 2 is shown in Figure 82.

[0523] TIFF2025526212000078.tif65170

[0524] Compound III Pattern 2 is an anhydride. The stoichiometry of the free base:fumaric acid is 1 The ratio is about 1:1.2 based on H NMR results. The T onset , and a T of 118.8°C with an enthalpy of approximately 63 J / g onset It has two melting peaks, and shows a weight loss of about 0.3% at about 105°C. The residual solvent is 1 Not detected by 1 H NMR.

[0525] Example 17: Preparation of Compound V Pattern 1 100mg of S PCompound I free base and 1.0 equivalent of fumaric acid were added to a glass vial, followed by 0.8 mL of IPA. After stirring at 50°C for approximately 1 hour, a clear solution was obtained. Approximately 2 mg of Compound III Pattern 1 seeds were added to the mixture. After observing the precipitation of some solid mass, 1 mL of heptane was added to the mixture. The mixture was stirred at 50°C for 2 hours and then cooled to 3°C at 0.1°C / min. It was maintained stirring at 3°C for approximately 3 days. The solid was isolated by filtration and dried in a vacuum oven at 50°C for approximately 2 hours to obtain Compound V Pattern 1. The results are reported in Table 42. The XRPD diffractogram of Compound V Pattern 1 is shown in Figure 83. The DSC thermogram of Compound V Pattern 1 is shown in Figure 84. The TGA thermogram of Compound V Pattern 1 is shown in Figure 85.

[0526] TIFF2025526212000079.tif55170

[0527] Example 18: Preparation of Compound V Pattern 2 100mg of S P Compound I free base and 1.0 equivalent of fumaric acid were added to a glass vial, followed by 0.8 mL of IPA. To the resulting clear solution, 1 mL of heptane was added, and the mixture was stirred at 50°C for 2 hours, then cooled to 3°C at 0.1°C / min. It was maintained under stirring at 3°C for approximately 3 days. The solid formed was isolated by filtration and dried in a vacuum oven at 50°C for approximately 2 hours to obtain Compound V Pattern 2. The results are reported in Table 43. The XRPD diffractogram of Compound V Pattern 2 is shown in Figure 86. The DSC thermogram of Compound V Pattern 2 is shown in Figure 87. The TGA thermogram of Compound V Pattern 2 is shown in Figure 88.

[0528] TIFF2025526212000080.tif60170

[0529] Example 19: Bulk Stability of Compound II Pattern 1 and Compound III Pattern 2 Compound II Pattern 1 and Compound III Pattern 2 were placed in an open container at 25°C / 92%RH, an open container at 40°C / 75%RH, and a closed container at 60°C for one week. The samples were characterized by XRPD and HPLC and examined for color change. The results are shown in Table 44.

[0530] TIFF2025526212000081.tif91170

[0531] Initial chemical and chiral purity The initial chemical purities of Compound II Pattern 1 and Compound III Pattern 2 are 99.7% and 98.8%, respectively. The chiral purity (% de) of Compound II Pattern 1 is 98.4%.

[0532] Bulk Stability Accelerated stability studies were conducted for one week in an open container at 25°C / 92% RH, an open container at 40°C / 75% RH, and an airtight container at 60°C. Compound II Pattern 1 showed good physical and chemical stability after exposure to the three conditions. Compound III Pattern 2 showed good physical stability under the above three conditions. Degradation products increased by 1.6% and 1.5% after exposure to 40°C / 75% RH in an open container and 60°C in a sealed container, respectively.

[0533] Example 20: Solubility Study of Compound II Pattern 1 and Compound III Pattern 2 12 mg of Compound II Pattern 1 and 12 mg of Compound III Pattern 2 were accurately weighed into an 8 mL glass vial and 5 mL of dissolution medium was added. The amount of salt used was equivalent to 10 mg of the anhydrous free base. All samples were clear solutions after 0.5 and 2 hours at 37°C in the medium. The pH of the resulting clear solutions was analyzed with a pH meter, and the solubility was determined by observation.

[0534] TIFF2025526212000082.tif72170

[0535] Solubility was tested in five media: pH 3.0 citrate buffer, pH 4.5 acetate buffer, pH 6.8 phosphate buffer, water, and simulated vaginal fluid (pH 4.2) for 0.5 and 2 hours at 37° C. Two candidates, Compound II Pattern 1 and Compound III Pattern 2, were highly soluble in the media (greater than 2 mg / mL).

[0536] Example 21: Hygroscopicity of Compound II Pattern 1 and Compound III Pattern 2 Hygroscopicity was investigated by DVS at 25°C using the following method. Method: 40-0-95-0-40%RH, dm / dt=0.002

[0537] Both Compound II Pattern 1 and Compound III Pattern 2 are slightly hygroscopic. Compound II Pattern 1 exhibits water absorption of approximately 0.2% up to 95% RH. No morphological changes were observed after the DVS test. Compound III Pattern 2 exhibits water absorption of approximately 1.0% up to 95% RH. No morphological changes were observed after the DVS test. The results are shown in Table 46.

[0538] TIFF2025526212000083.tif104170

[0539] Example 22: Polymorph Screening of Compound II Pattern 1 and Compound III Pattern 2 50 mg of the monofumarate was equilibrated in a suitable amount of solvent or solvent mixture. The resulting suspension was equilibrated for one week. The solid was isolated by centrifugal filtration. The wet cake obtained after equilibration was analyzed by XRPD to determine the crystal morphology change.

[0540] TIFF2025526212000084.tif243170TIFF2025526212000085.tif108170

[0541] Polymorphic evaluation of compound II pattern 1 In this study, no dissociation was observed during equilibration experiments, and two new potential polymorphs of the fumarate salt of Isomer I (Compound II Pattern 2 and Compound II Pattern 3) were obtained from acetonitrile and MEK, respectively, both of which exhibited lower melting temperatures than Compound II Pattern 1. Both polymorphs of Isomer 1 exhibited unchanged high chiral purity.

[0542] Polymorphic evaluation of compound III pattern 2 In this study, dissociation was observed during equilibration experiments to give the hemifumarate of isomer II (hemifumarate pattern 2). Additionally, four new monofumarates of isomer II (compound III pattern 3, compound III pattern 4, compound III pattern 5, and compound III pattern 6) were obtained.

[0543] Example 23: X-ray Powder Diffraction (XRPD) XRPD analysis was performed on a Bruker D8 Advance diffractometer.

[0544] Table 48 below provides the results of XRPD performed on Compound II Pattern 1. The XRPD showed sharp peaks, indicating that the sample was composed of crystalline material. The XRPD of Compound II Pattern 1 showed prominent peaks at about 3.1±0.2°, about 9.3±0.2°, about 12.1±0.2°, about 14.9±0.2°, about 15.1±0.2°, about 18.1±0.2°, about 19.8±0.2°, about 20.1±0.2°, about 25.1±0.2°, about 25.9±0.2°, and about 28.8±0.2°.

[0545] TIFF2025526212000086.tif137170

[0546] Table 49 below shows the results of XRPD performed on Compound IV Pattern 1. The XRPD of Compound IV Pattern 1 showed prominent peaks at about 6.5±0.2°, about 12.1±0.2°, about 17.5±0.2°, about 18.1±0.2°, about 18.5±0.2°, about 19.6±0.2°, about 19.8±0.2°, about 20.2±0.2°, about 20.6±0.2°, and about 21.3±0.2°.

[0547] TIFF2025526212000087.tif110170

[0548] Table 50 below provides the results of XRPD performed on Compound III Pattern 1. The XRPD of Compound III Pattern 1 showed prominent peaks at about 9.5±0.2°, about 11.7±0.2°, about 14.6±0.2°, about 17.5±0.2°, about 18.0±0.2°, about 20.0±0.2°, about 20.4±0.2°, about 22.3±0.2°, about 23.7±0.2°, and about 25.5±0.2°.

[0549] TIFF2025526212000088.tif116170

[0550] Table 51 below provides the results of XRPD performed on Compound III Pattern 2. The XRPD showed sharp peaks, indicating that the sample was composed of crystalline material. The XRPD of Compound III Pattern 2 showed prominent peaks at about 8.9±0.2°, about 9.9±0.2°, about 11.7±0.2°, about 12.1±0.2°, about 15.1±0.2°, about 17.9±0.2°, about 18.2±0.2°, about 19.9±0.2°, about 25.1±0.2°, about 29.6±0.2°, and about 38.1±0.2°.

[0551] TIFF2025526212000089.tif126170

[0552] Table 52 below shows the results of XRPD performed on Compound V Pattern 1. The XRPD of Compound V Pattern 1 showed prominent peaks at about 5.0±0.2°, about 7.2±0.2°, about 10.1±0.2°, about 12.1±0.2°, about 17.5±0.2°, about 17.9±0.2°, about 19.3±0.2°, about 22.0±0.2°, about 24.3±0.2°, about 25.1±0.2°, and about 26.3±0.2°.

[0553] TIFF2025526212000090.tif80170

[0554] Table 53 below shows the results of XRPD performed on Compound V Pattern 2. The XRPD of Compound V Pattern 2 showed prominent peaks at about 5.1±0.2°, about 6.9±0.2°, about 7.6±0.2°, about 10.2±0.2°, about 11.6±0.2°, about 12.1±0.2°, about 15.1±0.2°, about 17.6±0.2°, about 18.1±0.2°, about 18.7±0.2°, about 19.5±0.2°, about 19.8±0.2°, and about 25.1±0.2°.

[0555] TIFF2025526212000091.tif111170

[0556] Example 24: Single Crystal X-ray Diffraction (SC-XRD) Study of Compound II Pattern 1 Single crystals of compound II pattern 1 suitable for SC-XRD studies were obtained by temperature cycling experiments in MeOH. X-ray diffraction data were collected on a D8 Venture diffractometer equipped with a CMOS area detector at 170(2) K using Cu-Kα radiation (λ = 1.5418 Å); X-ray generator power: 50 kV, 1.4 mA, sample-to-area detector distance: 40 mm, exposure time: 150 s, resolution: 0.81, structure refinement: F 2 , Hydrogen donor sites: mixed. H atoms were treated with a mixture of independent and constrained refinements. X-ray diffraction and crystallographic data are shown in Table 54.

[0557] TIFF2025526212000092.tif211170

[0558] The crystalline form of Compound II Pattern 1 is monoclinic, R int = 5.7% and a final R1 [I>2σ(I)] = 7.4%. The crystalline form was free of solvent molecules. This crystalline form of Compound II Pattern 1 was found to correspond to the mono-fumaric acid salt of Isomer I with a free base to fumaric acid ratio of 1:1 and a Flack parameter (absolute structural parameter) of 0.16 (10). As shown in Figure 115, the protonated free base and fumarate anion are located at the N-terminal end of the single crystal structure. + The fumaric acid is linked via a (5)-H(5)····O(7) ionic bond. Proton transfer from the fumaric acid to the N(5)-nitrogen atom of the purine was observed.

[0559] Example 25. Tablet stability test Tablets were prepared using the excipients and excipient ratios shown in Column 1 of the following table. The tablets were then stored under the specified conditions, sampled periodically, and tested for purity by HPLC. Tablets made from Compound I monofumarate showed less degradation than tablets made from Compound I across the various formulations.

[0560] TIFF2025526212000093.tif51170

[0561] TIFF2025526212000094.tif70170

[0562] Example 26: Synthesis of a mixture of (R,S) and (S,S) ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate (Compound I) [ka]

[0563] Step 1: Preparation of diethyl-((2-(2-amino-6-chloro-9H-purin-9-yl)-ethoxy)-methyl)-phosphonate (3) [ka] 2-Amino-6-chloropurine 1 (50 g, 0.296 mol, 1 equiv.), CsCO (96.37 g, 0.296 mol, 1 equiv.), and DMF (250 mL) were charged to a dry reaction vessel under a N atmosphere at room temperature. Diethyl 2-chloroethoxymethylphosphonate 2 (74.85 g, 0.325 mol, 1.1 equiv.) was added dropwise with stirring at room temperature. The reaction mixture was stirred at 40°C–50°C for 0.5–1.5 h, heated to 60°C–70°C, stirred for 0.5–1.5 h, and then stirred at 75°C–85°C for 18–24 h. After the reaction temperature was brought to 20°C–30°C, the reaction mixture was filtered, and the resulting cake was washed with DMF (100 mL × 2). The combined filtrate was concentrated to half its volume below 70 °C, diluted with n-heptane (250 mL), and again concentrated to half its volume below 75 °C. The resulting solution was poured into DCM (1 L) and stirred at 20–30 °C for 20–40 min, followed by the addition of 10% aqueous Na2SO4 (approximately 100 mL). The resulting biphasic solution was stirred for 20–40 min, then filtered through diatomaceous earth, and the wet cake was washed with DCM (approximately 100 mL). The aqueous phase was separated from the filtrate, and the organic phase was washed again with 10% aqueous Na2SO4 (approximately 100 mL). The aqueous phase was combined by back-extraction with DCM (200–300 mL), and the combined organic phases were concentrated. The resulting crude product 3 was then purified by silica gel column chromatography using DCM and 1% MeOH in DCM. The product-containing fractions were combined, and the solvent was evaporated below 40 °C. The solid product 3 was treated by repeated dilution and concentration (up to 1 / 3 volume) with MTBE. The resulting slurry was then diluted with MTBE (400 mL–500 mL) and stirred at 40–50 °C for 4–6 h and at 15–25 °C for 8–15 h. The suspension was filtered, washed with MTBE, and dried at 35–40 °C for 15–20 h to give the desired product, diethyl-((2-(2-amino-6-chloro-9H-purin-9-yl)-ethoxy)-methyl)-phosphonate 3, in 43.4% isolated yield (48.66 g) with a purity of 91.8% by HPLC. 1H NMR (400 MHz, DMSO-d6), δppm: 8.08 (s, 1H), 6.91 (s, 2H), 4.24 (d, 2H, J= 8 Hz), 3.92 (m, 4H), 3.86 (q, 4H, J= 8Hz), 1.14 (t, 6H, J= 8 Hz). LCMS (m / z): 364.2 (MH+) and 366.2 (MH+).

[0564] Step 2: Preparation of ((2-(2-amino-6-chloro-9H-purin-9-yl)-ethoxy)-methyl)-phosphonic acid (4) [ka]

[0565] Diethyl-((2-(2-amino-6-chloro-9H-purin-9-yl)-ethoxy)-methyl)-phosphonate 3 (100 g, 0.275 mol) was added to a dry vessel containing DCM (1 L) under a N atmosphere, followed by 2,6-lutidine (147.33 g, 1.375 mol, 5 equiv.), and the temperature was adjusted to 0-5 °C. To this, TMSBr (167.47 g, 1.102 mol, 4.0 equiv.) was added dropwise and the mixture was stirred at 0-5 °C for 0.5-1 h and at 20-25 °C for 15-20 h. After adjusting the reaction temperature to 0-5 °C, 1144 g of 1 N aqueous NaOH solution was added dropwise. After maintaining the temperature at 20-30 °C for 1-2 h, the aqueous alkaline layer was separated and repeatedly washed with MTBE. The aqueous solution was acidified to pH 6-7 by dropwise addition of 2N aqueous HCl at 15-25°C, and MeOH (10 volumes) was charged. The resulting methanol solution was further acidified to pH 3-4 by dropwise addition of 2N aqueous HCl at 35-45°C. After seeding with product 4, the methanolic acidic solution was stirred at 35-45°C for 3-5 hours, further acidified to pH 1.5-2.5 by dropwise addition of 2N aqueous HCl, and stirred at 15-20°C for 11-20 hours. The resulting solid was isolated by filtration, washed with MeOH (2 × 100 mL), and dried at 45–55 °C for 20–30 h to give the desired product ((2-(2-amino-6-chloro-9H-purin-9-yl)-ethoxy)-methyl)-phosphonic acid 4) in 96.5% (84.4 g) isolated yield with a purity of 99.8% by HPLC. 1 H NMR (400 MHz, DMSO-d6), δ ppm: 8.1 (s, 1H), 6.92 (bs, 2H), 4.5-5.5 (bs, 2H), 4.22 (dd= t, 2H, J= 8 Hz), 3.84 (t, 2H, J= 8 Hz), 3.58 (t, 2H, J= 8 Hz). LCMS (m / z): 308 (MH+) and 310 (MH+).

[0566] Step 3: Preparation of ((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-phosphonic acid (5) [ka]

[0567] ((2-(2-amino-6-chloro-9H-purin-9-yl)-ethoxy)-methyl)-phosphonic acid 4 (50 g, 0.162 mol) was charged to a flask containing MeOH (350 mL) at 20-30 °C and stirred for 10-30 min. To this solution, a 30 wt% NaOMe solution (1.62 mol, 10 equiv.) in MeOH was added dropwise and then stirred at 50-60 °C for 15-24 h. The reaction was maintained at 20-30 °C for 20-40 min and then filtered. The filtrate was then acidified to pH 6-7 by dropwise addition of concentrated HCl at 20-30 °C and concentrated to one-third volume below 40 °C. The temperature of the concentrated solution was raised to 35-45 °C and acidified to pH 3-4 by dropwise addition of concentrated HCl. The resulting acidic solution was seeded with product 5 and stirred at 35°C to 45°C for 1.5 to 2.5 hours. At this temperature, the pH was adjusted to 2 to 3 by dropwise addition of concentrated HCl, stirred for 3 to 5 hours, cooled to -3°C to 3°C, and stirred for 8 to 15 hours. The resulting solid was filtered and washed with MeOH (approximately 100 mL) and n-heptane (approximately 100 mL). The resulting cake was dried under vacuum at 50°C to 60°C for 16 to 24 hours to afford the desired product ((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-phosphonic acid 5) in 89.3% isolated yield (48.22 g) with a purity of 99.5% by HPLC. 1 H NMR (400 MHz, DMSO-d6), δ, ppm: 7.88 (s, 1H), 6.47 (bs, 4H), 4.18 (t, 2H, J= 8 Hz), 3.96 (t, 2H, J= 8 Hz), 3.60 (d, 2H, J= 12 Hz). LCMS (m / z): 304.20 (MH+).

[0568] Step 4a: Preparation of a mixture of (R,S)- and (S,S)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate (8) [ka]

[0569] To a solution of ((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-phosphonic acid 5 (40 g, 0.132 mol, 1 equiv.) in DCM (560 mL) was charged (S)-ethyl 2-aminopropionate hydrochloride salt 6 (20.19 g, 0.132 mol, 1 equiv.), benzyl alcohol 7 (71.28 g, 0.66 mol, 5 equiv.), and TEA (159.98 g, 1.58 mol, 12 equiv.) with stirring at 20-30 °C, and the solution was stirred for 10-30 min. To this was added a solution prepared from Ph3P (207.5 g, 0.792 mol, 6 equiv.) and 2,2'-dithiopyridine (Aldrithiol-2) (174.24 g, 0.792 mol, 6 equiv.) in DCM (320 mL) over 60 min at 20-30 °C. The resulting reaction mixture was stirred at 35-45 °C for 15-20 h, concentrated to remove 3 / 4 of the solvent under vacuum below 40 °C, and the resulting residue was added with MeOH (approximately 120 mL), distilled water (approximately 400 mL), toluene (approximately 400 mL), and n-heptane (approximately 400 mL) and stirred at 20-30 °C for 0.5-1 h. After the reaction mixture was allowed to stand at 20-30°C for 0.5-1 hour, the organic phase was separated, and the aqueous phase was further extracted several times with a mixture of toluene (approximately 400 mL) and n-heptane (approximately 400 mL) to remove the maximum amount of remaining reagents and by-products. The remaining aqueous phase was then extracted with DCM (2 x 400 mL), and the DCM was concentrated under vacuum at <40°C. The crude product was purified by silica gel column chromatography using DCM to 2% MeOH in DCM as the mobile phase. The product-containing eluted fractions were combined and the solvent removed under reduced pressure below 40°C to give the desired product, a mixture of (R,S) and (S,S) diastereoisomers, i.e., (±)(2S)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate 8 (Compound I), in 45.8% (29.74 g) isolated yield, with a purity of 98.8% by HPLC. 1H NMR (400 MHz, DMSO-d6), δ, ppm: 7.85 (s, 1H), 7.34 (m, 5H), 6.44 (s, 2H), 5.36 (m, 1H), 4.90 (m, 2H), 4.17 (m, 2H), 4.07 (m, 2H), 3.95 (s, 3H), 3.82 (m, 5H), 1.18-1.24 (m, 6H). LCMS (m / z): 493.3 (MH+).

[0570] Step 4b: Preparation of a mixture of (R,R)- and (S,R)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate [ka]

[0571] To synthesize the (R,R) and (S,R) mixtures, the procedure in step 4a can be carried out by substituting D-alanine ethyl ester ((R)-ethyl 2-aminopropionate hydrochloride) for L-alanine ethyl ester ((S)-ethyl 2-aminopropionate hydrochloride).

[0572] Example 27: Preparation of (±)-Compound I monofumarate ((±)-(2S)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate monofumarate) (9) [ka]

[0573] To a solution of (±)-(2S)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate (compound I) 8 (29.72 g, 0.06 mol, 1 equiv.) in IPA, a solution of fumaric acid (7.66 mol, 1.1 equiv.) in IPA was added through a filter at 45-55°C, and stirring was continued for 1-2 h. Seeds of compound 9 were added to the reaction mixture, and stirring was continued at 45-55°C for 1-2 h. The reaction mixture was allowed to settle at 20-30°C for 4-6 h, after which n-heptane (approximately 300 mL) was added dropwise, and stirring was continued for an additional 8-15 h at 20-30°C and 8-15 h at 0-5°C. The observed solid was filtered, and the wet cake was washed with a mixture of IPA / n-heptane (1 / 3, vol / vol, approximately 50 mL-60 mL). The solid cake was dried under vacuum at 35-45 °C for 16-24 h to afford the desired product, (±)-(2S)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate monofumarate 9 (compound I monofumarate), in 87.9% (32.74 g) isolated yield with a purity of 99.1% by HPLC. 1 H NMR (DMSO-d6), δ, ppm: 1.14 (t, 3H, J= 7.2 Hz), 1.22 (d, 3H, J= 7.2 Hz), 3.82 (m, 2H; dd, 1H, J= 4.0 Hz; bs, 2H), 3.95 (s, 3H), 4.06 (m, 2H), 4.17 (m, 2H), 4.87 (m, 2H), 5.38 (q, 1H, J= 4 Hz), 6.44 (s, 2H), 6.64 (s, 2H), 7.33 (m, 5H), 7.82 (s, 1H), 13.18 (bs, 2H). LCMS (m / z): 493.20 (MH+).

[0574] Example 28: Chiral Separation of the (R,S)- and (S,S)-Isomers of Compound I and Preparation of Their Monofumarate Salts, Compound II and Compound III [ka]

[0575] Step 1a: Chiral separation of (R,S)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate and (S,S)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate Compound I (a diastereoisomeric mixture of isomers I and II) (22.50 g) was subjected to chiral chromatographic separation under the SFC separation conditions shown below to obtain 11.7 g of (R,S)-isomer I (10) with a purity of 98.6% by HPLC and 9.1 g of (S,S)-isomer II (11) with a purity of 95.6% by HPLC.

[0576] SFC conditions: Column: ChiralPak AD, 250 × 30 mm i.d., 10 μm; Mobile phase: A: CO2 and B: ethanol (0.1% NH3H2O); Gradient: B45% isocratic; Flow rate: 200mL / min; Wavelength: 310nm; Cycle time: about 6 minutes; Back pressure: 100bar; Injection amount: Approximately 1g.

[0577] Characterization of (R,S)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate (isomer I) as the free base: purity by HPLC: 98.6%; 1H NMR (DMSO-d6), δ, ppm: 7.82 (s, 1H), 7.30 (m, 5H), 6.38 (s, 2H), 5.30 (t, 1H), 4.83 (d, 2H), 4.18 (t, 2H), 4.05 (m, 2H), 3.95 (s, 3H), 3.84 (m, 2H), 3.60 (m, 5H), 1.20 (d, 3H), 1.15 (t, 3H); LCMS (m / z): 493 (MH+).

[0578] Characterization of (S,S)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate (isomer II) as the free base: purity by HPLC: 95.6%; 1 H NMR (DMSO-d6): δppm 7.82 (s, 1H), 7.35 (m, 5H), 6.45 (s, 2H), 5.30 (t, 1H), 4.80 (d, 2H), 4.18 (t, 2H), 4.05 (m, 2H), 3.95 (s, 3H), 3.80 (m, 3H), 3.70 (m, 2H), 1.20 (d, 3H), 1.15 (t, 3H); LCMS (m / z): 493 (MH+).

[0579] In certain non-limiting embodiments, stereoisomers are separated using HPLC or SFC with an achiral or chiral stationary phase. Non-limiting examples of chiral stationary phases that can be used include Chiralpak AD, Chiralpak AS, Chiralcel OG, and Chiralcel OJ.

[0580] In an alternative, non-limiting embodiment, individual isomers can be synthesized asymmetrically. For non-limiting examples of asymmetric synthesis of phosphonamidates, see Numan, A. et al. "Asymmetric Synthesis of Stereogenic Phosphorus P(V) Centers Using Chiral Nucleophilic Catalysis", Molecules 2021, 26, 3661 and Ambrosi, A. et al. "Synthesis of Rovafovir Etalafenamide (Part III): Evolution of the Synthetic Process to the Phosphonamidate Fragment", 2021, Org. Process Res. Dev. 25, 5, 1247-1262.

[0581] Step 1b: Chiral separation of (R,S)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate and (S,S)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate [ka]

[0582] Separation of (R,R)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate and (S,R)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methoxy)-(benzyloxy)-phosphoryl)-amino)-propionate (synthesis described in step 4b of Example 26) can be carried out using the same techniques as for the (R,S) and (S,S) mixtures described above.

[0583] Step 2a: Preparation of (R,S)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate monofumarate (Compound II) [ka]

[0584] To a solution of (R,S)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate 10 (3 g, 6 mmol, 1 equiv.) in IPA, a solution of fumaric acid (0.765 g, 6.6 mmol, 1.1 equiv.) in IPA was added through a filter at 45-55 °C, and stirring was continued for 1-2 h. Seeds of compound 12 were added to the reaction mixture, and stirring was continued at 45-55 °C for 1-2 h. The reaction mixture was allowed to settle at 20-30 °C for 4-6 h, after which n-heptane (approximately 30 mL) was added dropwise, and stirring was continued for another 8-15 h at 20-30 °C and 8-15 h at 0-5 °C. The observed solid was filtered, and the wet cake was washed with a mixture of IPA / n-heptane (1 / 3, v / v, ca. 5 mL). The solid cake was dried under vacuum at 35-45 °C for 16-24 h to give the desired product, (R,S)-ethyl-2((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate monofumarate 12 (isomer I monofumarate or compound II), in 85% (3.1 g) isolated yield with 98.6% purity by HPLC. 1H NMR (DMSO-d6), δ, ppm:δ7.80 (s, 1H), 7.35 (m, 5H), 6.63 (s, 2H), 6.40 (s, 2H), 5.53 (t, 1H), 4.84 (d, 2H), 4.15 (t, 2H), 4.00 (m, 2H), 3.92 (s, 3H), 3.80 (m, 3H), 3.75 (m, 2H), 1.20 (d, 3H), 1.13 (t, 3H); base (10):fumaric acid ratio = 1:1.00( 1 by H NMR).

[0585] Step 2b: Preparation of (S,S)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate monofumarate (13) (also referred to as Compound III) [ka]

[0586] To a solution of (R,S)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate 11 (3 g, 6 mmol, 1 equiv.) in IPA, a solution of fumaric acid (0.765 g, 6.6 mmol, 1.1 equiv.) in IPA was added through a filter at 45-55 °C, and stirring was continued for 1-2 h. Seeds of compound 13 were added to the reaction mixture, and stirring was continued at 45-55 °C for 1-2 h. The reaction mixture was allowed to settle at 20-30 °C for 4-6 h, after which n-heptane (approximately 30 mL) was added dropwise, and stirring was further continued at 20-30 °C for 8-15 h and at 0-5 °C for 8-15 h. The observed solid was filtered, and the wet cake was washed with a mixture of IPA / n-heptane (1 / 3, v / v, ca. 5 mL). The solid cake was dried under vacuum at 35-45 °C for 16-24 h to give the desired product, (S,S)-ethyl-2((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate monofumarate 13 (isomer II monofumarate or compound III), in 80% (2.9 g) isolated yield with a purity of 95.6% by HPLC. 1 H NMR (DMSO-d6), δ, ppm: δ7.82 (s, 1H), 7.35 (m, 5H), 6.62 (s, 2H), 6.35 (s, 2H), 5.30 (t, 1H), 4.90 (d, 2H), 4.15 (t, 2H), 4.05 (m, 2H), 3.95 (s, 3H), 3.80 (m, 3H), 3.70 (m, 2H), 1.20 (d, 3H), 1.15 (t, 3H); base (11):fumaric acid ratio = 1:1.2( 1 by H NMR).

[0587] Step 2c: Preparation of (R,R)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate monofumarate and (S,R)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate monofumarate [ka]

[0588] The synthesis of (R,R)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate monofumarate and (S,R)-ethyl-2-((((2-(2-amino-6-methoxy-9H-purin-9-yl)-ethoxy)-methyl)-(benzyloxy)-phosphoryl)-amino)-propionate monofumarate can be carried out in the same manner as was done in steps 2b and 2c for the (R,S) and (S,S) stereoisomers, replacing the starting materials in steps 2b and 2c with the product of the chiral separation in step 1b.

[0589] Example 29. Non-limiting example of preparation of semi-solid formulation A topical cream formulation can be prepared, for example, by emulsifying an oil phase and an aqueous phase together with an active pharmaceutical ingredient. In a non-limiting embodiment, the oil phase of the cream was prepared by mixing light mineral oil, propylparaben, and Tefose™ 63. Next, the aqueous phase of the cream was prepared by mixing water, EDTA, methylparaben, and Carbopol™ 974P. The oil phase and the aqueous phase were then emulsified. The active pharmaceutical ingredient and propylene glycol were added to the emulsified mixture. The pH of the mixture was adjusted and then filled into a tube.

[0590] The topical gel formulation can be prepared, for example, by mixing an aqueous gel carrier with an active pharmaceutical ingredient. In a non-limiting embodiment, the aqueous phase of the topical gel was prepared by mixing water, EDTA, methylparaben (or sorbic acid), and Carbopol™ 974P. The active pharmaceutical ingredient and propylene glycol were added to this solution, mixed, the pH was adjusted, and then the solution was filled into a tube.

[0591] In certain non-limiting embodiments, about 0.001% w / w to about 10% w / w of the active pharmaceutical ingredient is added to the semi-solid formulation, for example, about 0.0025% w / w to about 2.5% w / w, such as 0.003%, 0.01%, 0.03%, 0.1%, 0.3%, or 1%.

[0592] Example 30. Preparation of Compound I monofumarate tablets A non-limiting example of the preparation of Compound I monofumarate cervical tablets is provided below (see Figure 119 for a flow diagram). Two or more excipients are combined, blended, and screened to create an excipient blend. An active pharmaceutical ingredient (e.g., Compound I monofumarate) is then screened and added to a portion of the excipient blend. The resulting mixture is then blended, and then more excipient blend is added. In this way, the mixture is gradually diluted with the excipient blend, and thoroughly mixed after each addition of the excipient blend. When the excipient blend is exhausted, magnesium stearate is added, and the mixture is blended again. The mixture is then compressed into tablets and packaged.

[0593] TIFF2025526212000107.tif75170

[0594] Compound I monofumarate vaginal tablets 0.3mg A non-limiting example of a process for preparing vaginal tablets of Compound I monofumarate is provided below.

[0595] dispensing 1. Weigh out ingredients according to batch production formula and dispense into separate poly bags.

[0596] screening 1. Sift all excipients through a screen.

[0597] Active ingredient blending and screening 1. The screened excipients: microcrystalline cellulose and mannitol are blended in a diffusion blender. 2. Take 49.5 grams of excipient blend and add 2.12 grams of Compound I monofumarate. 3. The active ingredient and excipients are blended and screened to remove lumps. 4. To this blend, add 148.5 grams of excipient blend. 5. The active ingredient blend and excipients are blended and screened to remove lumps. 6. To this blend, add 247.5 grams of excipient blend. 7. The active ingredient blend and excipients are blended and screened to remove lumps. 8. To this blend, add the remaining 495 grams of excipient blend. 9. The active ingredient blend and excipients are blended and screened to remove lumps.

[0598] final mix 10. Add magnesium stearate to a diffusion blender and blend the contents. 11. Drain and blend.

[0599] compression 1. Compress the blend to target weight on a rotary tablet press using appropriate tooling (punches and dies). Check friability and disintegration at the start of the compression run and periodically check individual tablet weight, thickness and hardness.

[0600] packaging 1. Bulk tablets were packaged in a double-lined, reclosable transparent PE bag with a desiccant between the bags, and then placed in an aluminum foil pouch containing a desiccant and heat-sealed.

[0601] Example 31. Exemplary excipients for tablet formulations Tablet formulations are selected to exhibit mucoadhesive and substantive properties and include excipients with solubilizing, erodogenic (for disintegration), porosity (for water uptake), and viscosity-enhancing (to retain the drug at the target site) properties. Examples of excipients that cause rapid disintegration to coat the cervical, anal, or vaginal areas include, but are not limited to, mannitol, microcrystalline cellulose, lactose, sucrose, calcium phosphate, sodium phosphate, sodium bicarbonate, citric acid, maleic acid, adipic acid, or fumaric acid. Examples of excipients that can promote disintegration and coverage of the affected area include, but are not limited to, sodium starch glycolate, pregelatinized starch, crospovidone, and croscarmellose sodium. Mucoadhesive excipients useful in the present invention include, but are not limited to, microcrystalline cellulose, polycarbophil, hydroxymethylcellulose, hypromellose, hydroxypropylcellulose, and PVP.

[0602] The following table lists excipient combinations with desirable properties for a tablet formulation. The tablet formulation includes an active pharmaceutical ingredient, microcrystalline cellulose, and optionally mannitol. In certain non-limiting embodiments, the tablet formulation includes one or more excipients selected from the rapid disintegrant category (left column of Table 58). In certain non-limiting embodiments, the tablet formulation includes one or more excipients selected from the disintegration enhancing category (middle column of Table 58). In certain non-limiting embodiments, the tablet formulation includes one or more excipients selected from the mucoadhesive excipient category (right column of Table 58).

[0603] TIFF2025526212000108.tif169170

[0604] Example 32. Exemplary excipients for reconstituted powder or dry powder formulations Reconstituted powders or dry powder formulations can improve the storage stability of the drug or formulation. In certain non-limiting embodiments, the dry powder formulation can be mixed with saline, propylene glycol, or other aqueous carriers immediately before administration, minimizing degradation time. In certain non-limiting embodiments, the dry powder formulation is mixed with oil, cream, or other non-aqueous carrier immediately before administration.

[0605] In certain embodiments, the reconstituted powder or dry powder formulation rapidly coats infected or diseased tissues of the cervix, vulva, vagina, perianal area, penis, or anus. Excipients that enhance rapid coating of the cervix, vulva, vagina, perianal area, penis, or anus include, but are not limited to, mannitol, lactose, sucrose, calcium phosphate, and microcrystalline cellulose. In certain embodiments, the excipient for rapid coating of the cervix, vulva, vagina, perianal area, penis, or anus is mannitol.

[0606] In certain embodiments, the reconstituted powder or dry powder formulation has good coverage of the cervix, vulva, vagina, perianal area, penis, or anus. Non-limiting examples of excipients that enhance coverage of the cervix, vulva, vagina, perianal area, penis, or anus include sodium starch glycolate, pregelatinized starch, crospovidone, and croscarmellose sodium.

[0607] In certain embodiments, the reconstituted powder or dry powder formulation has mucoadhesive properties once reconstituted. This prevents smearing of the dosage form or exposure of healthy tissue to the active pharmaceutical ingredient. Excipients that improve the mucoadhesive properties of the reconstituted powder or dry powder formulation include, but are not limited to, xanthan gum, polycarbophil, polyethylene oxide, hydroxyethyl methylcellulose, hydroxyethyl cellulose, hypromellose, hydroxypropyl cellulose, PVP, and microcrystalline cellulose. In certain embodiments, the excipient that improves mucoadhesion is xanthan gum.

[0608] The following table lists excipient combinations with desirable properties for reconstitution powder or dry powder formulations. The dry powder or reconstitution powder formulation includes an active pharmaceutical ingredient and may include mannitol and / or xanthan gum. In certain non-limiting embodiments, the dry powder or reconstitution powder formulation includes one or more excipients selected from the rapid coating category (left column of Table 59). In certain non-limiting embodiments, the dry powder or reconstitution powder formulation includes one or more excipients selected from the enhanced coating category (middle column of Table 59). In certain non-limiting embodiments, the dry powder or reconstitution powder formulation includes one or more excipients selected from the mucoadhesive excipient category (right column of Table 58).

[0609] TIFF2025526212000109.tif169170

[0610] Example 33. Exemplary excipients for semi-solid formulations Semi-solid formulations are selected to exhibit mucoadhesive properties and aid in the penetration of drugs into tissues, and may include excipients with solubilizing, lipophilic (to aid in the solubilization of lipophilic compounds), penetration-enhancing (for higher activity), and mucoadhesive (to retain the drug at the target site) properties.

[0611] In certain embodiments, the semi-solid formulation is mucoadhesive. Excipients that contribute to mucoadhesion include, but are not limited to, carbomer, polyethylene glycol, crospovidone, polycarbophil, hypromellose, and hydroxyethylcellulose.

[0612] In certain embodiments, the semi-solid formulation enhances the permeability and / or solubility of the active pharmaceutical ingredient. Excipients that enhance the permeability and / or solubility of the active pharmaceutical ingredient include, but are not limited to, polyoxyl 6 stearate type I, ethylene glycol stearate, polyoxyl 32 stearate type I, and propylene glycol.

[0613] The following table lists excipient combinations with desirable properties for semi-solid formulations. The semi-solid formulation comprises an active pharmaceutical ingredient and one or more excipients from each column of Table 60. In certain non-limiting embodiments, the semi-solid formulation comprises one or more excipients selected from the mucoadhesive polymer category (left column of Table 60). In certain non-limiting embodiments, the tablet formulation comprises one or more excipients selected from the solubility and penetration enhancer category (second column of Table 60). In certain non-limiting embodiments, the semi-solid formulation comprises one or more excipients selected from the lipophilic solubilizer category (third column of Table 60). In certain non-limiting embodiments, the semi-solid formulation comprises one or more excipients selected from the penetration enhancer category (right column of Table 60).

[0614] TIFF2025526212000110.tif252170

[0615] Example 34. Exemplary excipients for semi-solid formulations The pessary and film-forming formulations are selected to be solid at room temperature but soften at body temperature to release the active pharmaceutical ingredient. This allows for easy handling and storage of the formulations and also allows for the desired tissue coverage of the cervix, vulva, vagina, perianal area, penis, or anus to be achieved. In non-limiting embodiments of the film-forming formulations, one or more excipients from the left column of Table 61 provide the desired properties. In non-limiting embodiments of the pessary formulations, one or more excipients from the right column of Table 61 provide the desired properties.

[0616] TIFF2025526212000111.tif101170

[0617] Example 35. Exemplary tablet formulations In certain non-limiting embodiments, the tablet dosage form formulation includes the ingredients in Table 62. In certain non-limiting embodiments, the tablet dosage form formulation includes the ingredients in Table 63. An exemplary process for combining these ingredients into a tablet dosage form is found in Example 29.

[0618] TIFF2025526212000112.tif72170

[0619] TIFF2025526212000113.tif50170

[0620] Example 36. Exemplary Semi-Solid Formulations In certain non-limiting embodiments, the cream semisolid dosage form formulation includes the ingredients in Table 64. In certain non-limiting embodiments, the gel semisolid dosage form formulation includes the ingredients in Table 65. An exemplary process for combining these ingredients into a cream or gel semisolid dosage form can be found in Example 27.

[0621] TIFF2025526212000114.tif68170

[0622] TIFF2025526212000115.tif103170

[0623] Example 37. Exemplary film-forming formulations Film dosage forms can be prepared by solvent casting or hot melt extrusion. For example, to prepare a film dosage form, the active pharmaceutical ingredient is dissolved in a solution of excipients and water. This solution is then optionally degassed, cast into a thin film, and dried in an oven. Film dosage forms can also be prepared by hot melt extrusion. In certain embodiments, the active pharmaceutical ingredient is mixed with the excipient(s) in a hopper. These ingredients are then mixed, milled, and kneaded into a uniform mixture. The mixture is heated until it flows and extruded through a die onto a roller, where it cools. In certain embodiments, the ingredients of the film dosage form are shown in Table 66.

[0624] TIFF2025526212000116.tif39170

[0625] Example 38. Exemplary dry powder or reconstituted powder formulations In cert...

Claims

1. formula: 【Chemical 1】 Compound.

2. formula: 【Chemistry 2】 or a pharmaceutically acceptable salt thereof.

3. formula: 【Chemistry 3】 or a pharmaceutically acceptable salt thereof.

4. formula: 【Chemistry 4】 The compound according to claim 2.

5. formula: 【Chemistry 5】 The compound according to claim 3.

6. Formula of isolated morphic form 【Chemistry 6】 wherein the isolated morphic form is characterized by an XRPD pattern comprising at least five 2-theta values selected from 3.08±0.2°, 9.30±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 20.14±0.2°, 25.14±0.2°, and 28.82±0.2°.

7. Formula of isolated morphic form 【Chemistry 7】 wherein the isolated morphic form is characterized by an XRPD pattern comprising at least seven 2-theta values selected from 9.53±0.2°, 10.04±0.2°, 11.60±0.2°, 14.57±0.2°, 17.22±0.2°, 17.50±0.2°, 20.04±0.2°, 20.36±0.2°, 22.34±0.2°, 23.73±0.2°, 25.48±0.2°, 26.06±0.2°, 27.38±0.2°, and 32.20±0.2°.

8. Formula of isolated morphic form 【Chemistry 8】 wherein the isolated morphic form is characterized by an XRPD pattern comprising at least seven 2-theta values selected from 8.94±0.2°, 9.89±0.2°, 9.91±0.2°, 11.66±0.2°, 12.11±0.2°, 15.13±0.2°, 17.85±0.2°, 18.15±0.2°, 19.90±0.2°, 20.38±0.2°, 22.94±0.2°, 25.09±0.2°, 26.54±0.2°, 26.90±0.2°, 27.38±0.2°, 28.28±0.2°, 28.95±0.2°, 29.64±0.2°, and 38.07±0.2°.

9. Formula of isolated morphic form 【Chemistry 9】 wherein the isolated morphic form is characterized by an XRPD pattern comprising at least five 2-theta values selected from 3.08±0.2°, 9.30±0.2°, 12.08±0.2°, 14.92±0.2°, 15.10±0.2°, 20.14±0.2°, 25.14±0.2°, and 28.82±0.2°.

10. A pharmaceutical composition comprising a compound according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier.

11. A pharmaceutical composition comprising the morphic form of any one of claims 6 to 9 in a pharmaceutically acceptable carrier.

12. 12. The pharmaceutical composition of claim 10 or 11, which is in a solid dosage form.

13. 12. The pharmaceutical composition of claim 10 or 11, which is in a semi-solid dosage form.

14. 12. The pharmaceutical composition of claim 10 or 11, in the form of a reconstituted powder.

15. 12. The pharmaceutical composition of claim 10 or 11, in the form of a dry powder dosage form.

16. 12. The pharmaceutical composition according to claim 10 or 11, in the form of a film.

17. 12. The pharmaceutical composition according to claim 10 or 11, in the form of a pessary.

18. 13. The pharmaceutical composition of claim 12 in the form of a tablet.

19. 14. The pharmaceutical composition of claim 13, in the form of a cream.

20. 14. The pharmaceutical composition of claim 13, in the form of a gel.

21. The pharmaceutical composition according to any one of claims 10 to 20, formulated for topical administration.

22. A pharmaceutical composition according to any one of claims 10 to 21 for delivery to the cervix.

23. 22. The pharmaceutical composition of any one of claims 10 to 21, for vaginal delivery.

24. A pharmaceutical composition according to any one of claims 10 to 21 for delivery to the vulva.

25. A pharmaceutical composition according to any one of claims 10 to 21 for delivery to the perianal area.

26. A pharmaceutical composition according to any one of claims 10 to 21, for delivery to the anus.

27. A pharmaceutical composition according to any one of claims 10 to 21 for delivery to the penis.

28. 19. The pharmaceutical composition of claim 18, wherein the tablet is a bilayer tablet.

29. 20. The pharmaceutical composition of claim 18, wherein the tablet disintegrates in less than about 250 μL of fluid.

30. 20. The pharmaceutical composition of claim 18, wherein the tablet disintegrates in less than about 150 μL of fluid.

31. 31. The pharmaceutical composition of any one of claims 10 to 30, comprising from about 0.01 mg to about 10 mg of the compound.

32. 31. The pharmaceutical composition of any one of claims 10 to 30, comprising from about 0.01 mg to about 5 mg of the compound.

33. 31. The pharmaceutical composition of any one of claims 10 to 30, comprising from about 0.03 mg to about 1 mg of the compound.

34. 31. The pharmaceutical composition of any one of claims 10 to 30, comprising about 0.03 mg to about 0.07 mg of the compound.

35. 31. The pharmaceutical composition of any one of claims 10 to 30, comprising about 0.05 mg to about 0.15 mg of the compound.

36. 31. The pharmaceutical composition of any one of claims 10 to 30, comprising from about 0.15 mg to about 0.45 mg of the compound.

37. 31. The pharmaceutical composition of any one of claims 10 to 30, comprising at least 0.05 mg of the compound.

38. 31. The pharmaceutical composition of any one of claims 10 to 30, comprising at least about 0.1 mg of the compound.

39. 31. The pharmaceutical composition of any one of claims 10 to 30, comprising from about 0.001% to about 10% of said compound.

40. 31. The pharmaceutical composition of any one of claims 10 to 30, comprising about 0.01% to 0.5% of said compound.

41. 31. The pharmaceutical composition of any one of claims 10 to 30, comprising about 0.1% to 5% of said compound.

42. The pharmaceutical composition according to any one of claims 10 to 41, comprising a mucoadhesive polymer.

43. 43. The pharmaceutical composition of claim 42, comprising about 5% to about 20% of a mucoadhesive polymer.

44. 43. The pharmaceutical composition of claim 42, comprising about 10% to about 50% mucoadhesive polymer.

45. 43. The pharmaceutical composition of claim 42, comprising about 50% to about 90% mucoadhesive polymer.

46. 42. The pharmaceutical composition of any one of claims 10 to 41, comprising a disintegration-enhancing excipient.

47. 42. The pharmaceutical composition of any one of claims 10 to 41, comprising a penetration enhancing excipient.

48. A pharmaceutical composition according to any one of claims 10 to 41, comprising excipients allowing a controlled release of the active compound.

49. 20. The pharmaceutical composition of claim 19, wherein the pharmaceutically acceptable carrier is comprised of light mineral oil, propylparaben, Tefose 63, water, EDTA, methylparaben, and Carbopol 974P.

50. 21. The pharmaceutical composition of claim 20, wherein the pharmaceutically acceptable carrier is comprised of water, EDTA, methylparaben, Carbopol 974P, propylene glycol, and sorbic acid.

51. 19. The pharmaceutical composition of claim 18, wherein the tablet is composed of mannitol, polycrystalline cellulose, and magnesium stearate.

52. 10. A method for treating a human papillomavirus infection, comprising administering to a host in need thereof an effective amount of a compound according to any one of claims 1 to 5, optionally in a pharmaceutically acceptable carrier.

53. A method for treating a condition caused by human papillomavirus infection, comprising administering to a host in need thereof an effective amount of a compound according to any one of claims 1 to 5, optionally in a pharmaceutically acceptable carrier.

54. 54. The method of claim 53, wherein the condition caused by human papillomavirus infection is an intraepithelial neoplasia.

55. 55. The method of claim 54, wherein the condition caused by human papillomavirus is atypical squamous cells of undetermined significance (ASC-US).

56. 55. The method of claim 54, wherein the condition caused by human papillomavirus is atypical glandular cells (AGC).

57. 55. The method of claim 54, wherein the condition caused by human papillomavirus is a low-grade squamous intraepithelial lesion (LSIL).

58. 55. The method of claim 54, wherein the condition caused by human papillomavirus is atypical squamous cells and high-grade squamous intraepithelial lesion cannot be excluded (ASC-H).

59. 55. The method of claim 54, wherein the condition caused by human papillomavirus is high-grade squamous intraepithelial lesion (HSIL).

60. 55. The method of claim 54, wherein the condition caused by human papillomavirus is adenocarcinoma occult (AIS).

61. 55. The method of claim 54, wherein the intraepithelial neoplasia is a cervical intraepithelial neoplasia.

62. 62. The method of claim 61, wherein the cervical intraepithelial neoplasia is grade 1 cervical intraepithelial neoplasia.

63. 62. The method of claim 61, wherein the cervical intraepithelial neoplasia is grade 2 cervical intraepithelial neoplasia.

64. 62. The method of claim 61, wherein the cervical intraepithelial neoplasia is grade 3 cervical intraepithelial neoplasia.

65. 55. The method of claim 54, wherein the intraepithelial neoplasia is a vaginal intraepithelial neoplasia.

66. 55. The method of claim 54, wherein the intraepithelial neoplasia is a vulvar intraepithelial neoplasia.

67. 55. The method of claim 54, wherein the intraepithelial neoplasia is an anal intraepithelial neoplasia.

68. 55. The method of claim 54, wherein the intraepithelial neoplasia is a perianal intraepithelial neoplasia.

69. 55. The method of claim 54, wherein the intraepithelial neoplasia is a penile intraepithelial neoplasia.

70. 70. The method of any one of claims 52 to 69, wherein the host is a human.

71. 71. The method of any one of claims 52 to 70, wherein the compound is administered topically.

72. 72. The method of any one of claims 52-71, wherein about 0.01 milligrams to about 1 milligram of the compound is administered.

73. 72. The method of any one of claims 52-71, wherein about 0.05 milligrams to about 0.3 milligrams is administered.

74. 74. The method of any one of claims 52 to 73, further comprising applying a lubricating means to epithelial tissue prior to inserting the dosage form into the affected area.

75. 74. The method of any one of claims 52 to 73, further comprising applying a lubricating means to the dosage form prior to inserting the dosage form into the affected area.

76. 76. The method of claim 74 or 75, wherein the lubricating agent is selected from water, glycerol-based lubricants, and hydroxyethylcellulose-based lubricants.

77. 77. The method of any one of claims 52 to 76, wherein the compound is administered daily.

78. 77. The method of any one of claims 52-76, administered about once daily.

79. 77. The method of any one of claims 52 to 76, administered about twice a week.

80. 77. The method of any one of claims 52-76, administered about three or more times per week.

81. 81. The method of any one of claims 52 to 80, wherein the method is administered for about one week.

82. 81. The method of any one of claims 52 to 80, wherein the method is administered for about two weeks.

83. 81. The method of any one of claims 52 to 80, wherein the method is administered for about 3 weeks.

84. 81. The method of any one of claims 52 to 80, wherein the method is administered for about 4 weeks.

85. 81. The method of any one of claims 52 to 80, wherein the method is administered for about 5 weeks.

86. 81. The method of any one of claims 52 to 80, wherein the method is administered for about 6 weeks.

87. The compound is a. a treatment cycle comprising administration of the compound; and b. A drug-free cycle, including a treatment-free period; 87. The method of any one of claims 52 to 86, wherein the method is administered in a treatment cycle comprising:

88. 88. The method of claim 87, wherein the drug holiday cycle is about one week.

89. 88. The method of claim 87, wherein the drug holiday cycle is about 2 weeks.

90. 88. The method of claim 87, wherein the drug holiday cycle is about 3 weeks.

91. 88. The method of claim 87, wherein the drug holiday cycle is about 4 weeks.

92. 88. The method of claim 87, wherein the drug holiday cycle is about 5 weeks.

93. 88. The method of claim 87, wherein the drug holiday cycle is about 6 weeks.

94. 94. The method of any one of claims 87 to 93, wherein at least two treatment cycles are applied.

95. 94. The method according to any one of claims 87 to 93, wherein at least three treatment cycles are applied.

96. 81. The method of any one of claims 52-80, wherein about 0.05 mg to about 0.3 mg of the compound is administered daily.

97. 81. The method of any one of claims 52-80, wherein about 0.05 mg to about 0.3 mg of the compound is administered three times per week.

98. 81. The method of any one of claims 52-80, wherein about 0.05 mg to about 0.3 mg of the compound is administered once a week.

99. 99. The method of any one of claims 52 to 98, wherein the human papillomavirus is a high-risk strain.

100. The method of any one of claims 52 to 98, wherein the human papillomavirus is HPV-16 or HPV-18.

101. 101. The method of any one of claims 52 to 100, wherein the compound is administered in combination with another antiviral compound.

102. 102. The method of claim 101, wherein the antiviral compound is selected from the group consisting of a protease inhibitor, another DNA polymerase inhibitor, an inhibitor of E6 or E6AP, an inhibitor of E7, an inhibitor of E1, an inhibitor of E2, an inhibitor of E1-E2 protein interaction, an L2 lipopeptide, an inhibitor of L1, an inhibitor of L2, a degrader of L1, and a degrader of L2.

103. 101. The method of any one of claims 52 to 100, wherein the compound is administered in combination with an anti-cancer compound.

104. 104. The method of claim 103, wherein the anti-cancer compound is selected from the group consisting of HDAC inhibitors, degraders of tetraspanins, immune checkpoint inhibitors, T-cell therapy, and anti-proliferative agents.

105. 101. The method of any one of claims 52 to 100, wherein the compound is administered in combination with a surgical procedure.

106. 106. The method of claim 105, wherein the compound is administered prior to the surgical procedure.

107. 106. The method of claim 105, wherein the compound is administered after the surgical procedure.

108. 106. The method of claim 105, wherein the surgical procedure is performed during administration of the compound.

109. 109. The method according to any one of claims 105 to 108, wherein the surgical procedure is the resection of diseased tissue.

110. 110. The method of claim 109, wherein the ablation is a loop electrosurgical ablation procedure (LEEP).

111. 110. The method of claim 109, wherein the resection is a large loop resection of the transition zone (LLETZ).

112. 110. The method of claim 109, wherein the excision is a knife conization.

113. 110. The method of claim 109, wherein the excision is laser conization.

114. 109. The method of any one of claims 105 to 108, wherein the surgical procedure is ablation of diseased tissue.

115. 115. The method of claim 114, wherein the ablation is laser ablation.

116. 115. The method of claim 114, wherein the ablation is cryoablation.

117. 10. Use of a compound according to any one of claims 1 to 5, optionally in a pharmaceutically acceptable carrier, in the manufacture of a medicament for the treatment of a human papillomavirus infection in a host in need thereof.

118. 10. Use of a compound according to any one of claims 1 to 5, optionally in a pharmaceutically acceptable carrier, in the manufacture of a medicament for the treatment of a condition caused by a human papillomavirus infection in a host in need thereof.

119. 119. The use of claim 118, wherein the condition caused by human papillomavirus infection is an intraepithelial neoplasia.

120. 120. The use of claim 119, wherein the intraepithelial neoplasia is a vaginal intraepithelial neoplasia.

121. 120. The use of claim 119, wherein the intraepithelial neoplasia is a vulvar intraepithelial neoplasia.

122. 120. The use of claim 119, wherein the intraepithelial neoplasia is a cervical intraepithelial neoplasia.

123. 120. The use of claim 119, wherein the intraepithelial neoplasia is an anal intraepithelial neoplasia.

124. 120. The use of claim 119, wherein the intraepithelial neoplasia is a perianal intraepithelial neoplasia.

125. 120. The use of claim 119, wherein the intraepithelial neoplasia is a penile intraepithelial neoplasia.

126. The use according to any one of claims 117 to 125, wherein the host is a human.

127. 127. The use according to any one of claims 117 to 126, for topical administration.

128. 6. A compound according to any one of claims 1 to 5, optionally in a pharmaceutically acceptable carrier, for use in the treatment of a human papillomavirus infection in a host in need thereof.

129. 6. A compound according to any one of claims 1 to 5, optionally in a pharmaceutically acceptable carrier, for use in the treatment of a condition caused by a human papillomavirus infection in a host in need thereof.

130. 130. The compound for use according to claim 129, wherein the condition caused by human papillomavirus infection is an intraepithelial neoplasia.

131. 131. The compound for use according to claim 130, wherein the intraepithelial neoplasia is a vaginal intraepithelial neoplasia.

132. 131. The compound for use according to claim 130, wherein the intraepithelial neoplasia is a vulvar intraepithelial neoplasia.

133. 131. The compound for use according to claim 130, wherein the intraepithelial neoplasia is a cervical intraepithelial neoplasia.

134. 131. The compound for use according to claim 130, wherein the intraepithelial neoplasia is an anal intraepithelial neoplasia.

135. 131. The compound for use according to claim 130, wherein the intraepithelial neoplasia is a perianal intraepithelial neoplasia.

136. 131. The compound for use according to claim 130, wherein the intraepithelial neoplasia is a penile intraepithelial neoplasia.

137. The compound for use according to any one of claims 128 to 136, wherein the host is a human.

138. 138. The compound for use according to any one of claims 128 to 137, wherein the compound is administered topically.

139. a.R Ｐ Dissolving Compound I in a solvent selected from methanol, ethanol, and isopropanol; b. stirring at a temperature of about 20°C to about 70°C; c. adding 1.0 equivalent of fumaric acid; d. adding a solvent selected from pentane, hexane, and heptane; e. Cooling the mixture; f. Stirring the cooled solution; g. isolating and drying the solid; 5. A method for preparing the morphic form of claim 4, comprising:

140. 140. The method of claim 139, wherein the alcohol solvent in step (a) is ethanol or isopropanol.

141. 141. The method of claim 139 or 140, wherein the alcohol solvent in step (a) is isopropanol.

142. 140. The method of claim 139, wherein the solution of step (b) is stirred at about 45°C to about 55°C.

143. 140. The method of claim 139, wherein the aliphatic solvent is hexane or heptane.

144. 144. The method of claim 139 or 143, wherein the aliphatic solvent is heptane.

145. 140. The method of claim 139, wherein the mixture is cooled to below about 20°C.

146. 140. The method of claim 139, wherein the mixture is cooled to less than about 10°C.

147. 140. The method of claim 139, wherein the mixture is cooled to less than about 5°C.

148. 140. The method of claim 139, wherein the mixture is cooled to about 5°C to 0°C.

149. 12. The pharmaceutical composition of claim 10 or 11, comprising microcrystalline cellulose.

150. 12. The pharmaceutical composition of claim 10 or 11, comprising lactose.

151. 12. The pharmaceutical composition of claim 10 or 11, comprising sucrose.

152. 12. The pharmaceutical composition of claim 10 or 11, comprising calcium phosphate.

153. 12. The pharmaceutical composition of claim 10 or 11, comprising sodium bicarbonate.

154. 12. The pharmaceutical composition of claim 10 or 11, comprising citric acid.

155. 12. The pharmaceutical composition according to claim 10 or 11, comprising sodium starch glycolate.

156. 12. The pharmaceutical composition according to claim 10 or 11, comprising pregelatinized starch.

157. 12. The pharmaceutical composition of claim 10 or 11, comprising crospovidone.

158. 12. The pharmaceutical composition of claim 10 or 11, comprising croscarmellose sodium.

159. 12. The pharmaceutical composition of claim 10 or 11, comprising polycarbophil.

160. 12. The pharmaceutical composition of claim 10 or 11, comprising polyethylene oxide.

161. 12. The pharmaceutical composition of claim 10 or 11, comprising hydroxyethyl methylcellulose.

162. 12. The pharmaceutical composition of claim 10 or 11, comprising hydroxyethylcellulose.

163. 12. The pharmaceutical composition according to claim 10 or 11, comprising hypromellose.

164. 12. The pharmaceutical composition according to claim 10 or 11, comprising maleic acid.

165. 12. The pharmaceutical composition according to claim 10 or 11, comprising adipic acid.

166. 12. The pharmaceutical composition according to claim 10 or 11, comprising fumaric acid.

167. 12. The pharmaceutical composition of claim 10 or 11, comprising hydroxypropyl cellulose.

168. 12. The pharmaceutical composition according to claim 10 or 11, comprising PVP.

169. 12. The pharmaceutical composition of claim 10 or 11, comprising xanthan gum.

170. 12. The pharmaceutical composition according to claim 10 or 11, comprising mannitol.

171. 12. The pharmaceutical composition of claim 10 or 11, comprising a carbomer.

172. 12. The pharmaceutical composition according to claim 10 or 11, comprising polyethylene glycol.

173. 12. The pharmaceutical composition according to claim 10 or 11, comprising cetyl alcohol.

174. 12. The pharmaceutical composition of claim 10 or 11, comprising stearyl alcohol.

175. 12. The pharmaceutical composition of claim 10 or 11, comprising a polysorbate.

176. 12. The pharmaceutical composition of claim 10 or 11, comprising sodium lauryl sulfate.

177. 12. A pharmaceutical composition according to claim 10 or 11, comprising a light mineral oil or mineral oil.

178. 12. The pharmaceutical composition according to claim 10 or 11, comprising a white wax.

179. 12. The pharmaceutical composition of claim 10 or 11, comprising a silicone fluid.

180. 12. A pharmaceutical composition according to claim 10 or 11, comprising transcutol.

181. 12. The pharmaceutical composition according to claim 10 or 11, comprising oleic acid.

182. 12. The pharmaceutical composition of claim 10 or 11, comprising isopropyl myristate.

183. 12. The pharmaceutical composition of claim 10 or 11, comprising propylene glycol dicaprylate.

184. 12. The pharmaceutical composition of claim 10 or 11, comprising glyceryl monooleate.

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