Pharmaceutical composition for herpes viruses

Abstract

The present invention relates to methods and compositions for treating and / or inhibiting the onset or progression of diseases or disorders caused by or associated with herpesvirus infection. In particular, long-acting injectable depot pharmaceutical compositions containing a helicase-primase inhibitor, methods for their preparation, and uses of the pharmaceutical compositions as medicines and for treating diseases or disorders caused by or associated with herpesvirus are provided.

Description

[Technical Field]

[0001] Provided herein are compositions and methods for treating and / or inhibiting the onset or progression of diseases or disorders caused by or associated with herpesvirus infection. In particular, provided herein are long-acting injectable depot pharmaceutical compositions containing a helicase-primase inhibitor, methods for their preparation, and uses of the pharmaceutical compositions as medicines and for the treatment of diseases or disorders caused by or associated with herpesvirus. [Background technology]

[0002] Human herpesviruses are large, enveloped, double-stranded DNA viruses characterized by the ability to establish lifelong infections in humans. This is achieved by their ability to exist in the host either as an asymptomatic latent infection (in which the virus is dormant) or as a lytic infection with associated symptoms after activation. These viral infections are widespread worldwide, and it is noteworthy that more than 90% of all humans are chronically infected with multiple human herpesviruses.

[0003] Human herpesviruses are classified into three subfamilies (α, β, and γ) based on their biological characteristics and consist of eight members: herpes simplex virus subtypes 1 and 2 (HSV1, HSV2), varicella-zoster virus (VZV), Epstein-Barr virus (EBV), cytomegalovirus (CMV), and human herpesviruses 6 to 8 (HHV6-8).

[0004] Both HSV1 and HSV2 infections can cause disease in immunocompetent individuals. Both subtypes cause cutaneous genital / anal and labial / nasal (cold sore) lesions, but HSV2 is more commonly associated with the former, while HSV1 is more commonly associated with the latter. It is believed that >80% of genital infections are caused by HSV2. Globally, over 500 million people suffer from genital herpes infections, and approximately 50-80% of the world's population suffers from oral HSV infections, which are the primary cause of cold sores. HSV, particularly HSV1, can also cause lesions on the hands (whitlow) and other parts of the skin.

[0005] The majority of people infected with HSV experience no noticeable symptoms. However, some experience recurrent (and often severe) outbreaks of infection. In the United States, 20–40% of the population has recurrent HSV lip lesions. Importantly, oral cold sores and felon provide a very easy route for viral transmission to others, which can lead to rarer but much more serious HSV-related conditions. For example, HSV-associated bulbar keratitis is a leading cause of blindness, and HSV can also cause encephalitis in newborns, a life-threatening condition. Other diseases thought to be caused by HSV include herpes gladiatorum, Mollaret's meningitis, and possibly Bell's palsy.

[0006] Primary infection or reactivation of pre-existing herpesvirus infection can be a major cause of disease in immunocompromised individuals. Major risk populations include those undergoing solid organ or stem cell transplantation, those undergoing cancer treatment, those with HIV / AIDS, and intensive care unit (ICU) patients.

[0007] There is currently no cure for HSV, and although drugs have been developed that can somewhat suppress outbreaks and / or shorten the duration of outbreaks, improved treatments are needed.

[0008] Nucleoside analogs, such as acyclovir and its prodrugs, valacyclovir and famciclovir, are currently used as drugs against herpes viruses, including HSV. To exert their effects, these nucleoside analogs must be phosphorylated by viral thymidine kinase (TK) and then converted by cellular kinases to nucleoside triphosphates that inhibit the activity of viral DNA polymerase. If the virus does not have a functionally active TK, such as in the case of resistant HHV1 mutants or TK-negative viruses, the nucleoside analogs cannot exert their effects.

[0009] Nucleoside analogs are administered clinically at very high doses, typically hundreds of milligrams to several grams per day. Even at these high doses, often administered over long periods of time, these drugs cannot completely prevent recurrent outbreaks of HSV symptoms. Nucleoside analogs also do little to address the problem of viral shedding, which can asymptomatically promote HSV infection in more individuals. Some nucleoside analogs, especially when used at high doses, raise safety concerns. For example, these drugs can be incorporated into host genomic DNA via host DNA polymerases, raising concerns about their mutagenicity, as documented for the nucleoside analog ganciclovir (Aoki, Chapter 45 in Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases (Eighth Edition) 2015).

[0010] Given the inadequacies of existing treatments, there is an urgent medical need to develop improved and well-tolerated anti-herpes treatments.

[0011] One class of compounds currently being investigated are helicase-primase inhibitors. Helicase-primase inhibitors are antiviral agents with a novel mechanism of action. They inhibit the viral heterotrimeric complex, which consists of a helicase subunit, a primase subunit, and a coenzyme subunit, a function essential for viral DNA replication. These agents are not nucleoside analogs and do not require phosphorylation by TK to inhibit HSV replication. Therefore, they may be active against TK-deficient HSV, which, as noted above, is a major mechanism of resistance to nucleoside analogs.

[0012] Two examples of helicase-primase inhibitors are BILS-179 BS (Crute et al., (2002) Nature Medicine 8, pp. 386-391) and amenamevir (Katsumata et al. (2018) Biochem Pharm 158 pp. 201-206).

[0013] BILS-179 BS has been administered orally to humans, but early clinical trials were discontinued due to adverse events (Ruebsamen et al., (2019) Med. Chem. Commun., DOI: 10.1039 / C9MD00233B). Similarly, amenamevir, which has been administered orally to humans, was discontinued in early clinical trials for HSV due to adverse events. In a dose-ranging, placebo-controlled study in 437 patients with recurrent genital herpes, amenamevir was administered orally at one of four doses: 100 mg, 200 mg, 400 mg, and 1200 mg. The primary endpoint of the study, time to lesion healing, was significantly different only between the highest single-dose 1200 mg group and the placebo group (Aoki, Chapter 45 in Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases (Eighth Edition) 2015).

[0014] Another example of a helicase-primase inhibitor is plitelivir, a thiazolylamide derivative with the chemical name N-methyl-N-(4-methyl-5-sulfamoyl-1,3-thiazol-2-yl)-2-[4-(pyridin-2-yl)phenyl]acetamide, which is disclosed in WO 2001 / 47904.

[0015] In a human clinical trial, the efficacy of pritelivir in suppressing genital herpes was studied in 156 people (Wald et al., (2014) New England Journal of Medicine 370, pp. 201-210). Subjects received one of three oral daily doses or one weekly oral dose of pritelivir or a placebo for 28 days. The four pritelivir dosing regimens were a 20 mg loading dose followed by a 5 mg daily dose; a 100 mg loading dose followed by a 25 mg daily dose; a 300 mg loading dose followed by a 75 mg daily dose; and a 400 mg weekly dose. At higher doses, pritelivir was found to reduce genital HSV shedding, the primary endpoint of the study. Pritelivir also reduced the number of disease-free days in healthy men and women with genital herpes. The 75 mg oral daily dose showed the greatest antiviral effect and was found to be superior to the less frequent 400 mg once weekly dose.

[0016] As highlighted in the study, the highest daily dose of 75 mg reduced HSV shedding rates compared to placebo, but breakthrough shedding persisted. Discussing this finding, Wald et al. ((2014) New England Journal of Medicine 370, pp. 201-210) noted that persistent, low-level shedding has also been observed with nucleoside therapy and that the etiology of breakthrough viral shedding during adequate antiviral therapy with nucleoside analogs is poorly understood; it is not related to poor adherence to the treatment regimen or viral resistance. The authors also questioned whether further increases in the daily dose of pritelivir would completely eliminate viral shedding, a question that should be addressed in future clinical trials.

[0017] Following this suggestion, subsequent clinical trials using even higher oral daily doses were indeed conducted. In a Phase II 28-day clinical trial involving 91 patients with recurrent genital HSV-2, HSV was shed in 5.3% of swabs after daily administration of 500 mg of valacyclovir, whereas HSV was shed in 2.4% of analyzed genital swabs after daily oral administration of 100 mg of pritelivir (after a loading dose of 400 mg). The results of this study were reported in Wald et al. (2016) (J. Am. Med. Assoc. 316(23), pp. 2495-2503), which noted that the daily dose of 100 mg of pritelivir (after a loading dose of 400 mg) was selected based on previous study results showing the high efficacy of a daily dose of 75 mg in suppressing viral shedding.

[0018] A current phase III clinical trial is investigating an oral loading dose of 400 mg pritelivir followed by oral pritelivir 100 mg daily for up to 28 days or until healing of all mucocutaneous HSV lesions, whichever occurs first (https: / / www.clinicaltrials.gov / ct2 / show / NCT03073967?term=pritelivir+f&draw=2&rank=2).

[0019] As is evident from the above, the primary route of administration investigated for all of the above-mentioned small molecule antiviral therapeutics is oral. Efforts to improve therapy, e.g., to further reduce or prevent HSV shedding and viral reactivation, have focused primarily on the use of higher oral doses (including higher loading doses) and more frequent administration (e.g., multiple daily doses).

[0020] The focus on increasing systemic exposure via the oral route of administration is evident in the various attempts to improve the oral bioavailability of nucleoside analogs and helicase-primase inhibitors. For example, valacyclovir was developed as a prodrug of acyclovir to provide higher oral bioavailability; the human bioavailability of valacyclovir after oral administration is approximately 54%, compared with only 10–20% for oral acyclovir (Murray (1995) Antiviral Chem. Chemotherapy 6(1), pp. 34–38). For pritelivir, efforts have focused on developing salt forms with improved solubility for oral delivery. For example, WO 2013 / 045491 discloses that improved oral bioavailability can be achieved by using a crystalline monomesylate monohydrate salt of pritelivir with specific particle characteristics. The publication states that the free base of pritelivir has poor solubility compared to the monomesylate monohydrate salt, resulting in unfavorable drug release and resorption characteristics. Furthermore, the free base is also characterized as being unsuitable for long-term stable formulations. International Publication No. 2013 / 045479 discloses a specific crystalline monomesylate monohydrate salt of pritelivir, and reports that this salt form has superior long-term stability after oral administration and improved systemic exposure compared to pritelivir free base.

[0021] There is a continuing need for new and improved methods for treating HSV infections, and it is with this in mind that the present invention has been devised. Summary of the Invention

[0022] In a first aspect, the present invention provides a long-acting injectable depot pharmaceutical composition comprising a helicase-primase inhibitor or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients, wherein the helicase inhibitor has a human biological terminal half-life of 10 hours or more.

[0023] Surprisingly, it has been found that when a helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, having a human biological terminal half-life of 10 hours or more is formulated as a long-acting injectable depot pharmaceutical composition according to the present invention, such a composition provides beneficial properties including specific release characteristics and profiles that allow plasma levels to be achieved over an extended period of time.

[0024] Surprisingly, in some embodiments, the long-acting injectable depot compositions of the present invention can provide steady, continuous release of a helicase-primase inhibitor for at least 14 days after administration. This allows for the possibility of less frequent administration and / or lower doses than other routes of administration. This is an unexpected finding. As noted above, oral administration is the primary route of administration under consideration for small molecule antiviral therapeutics, and efforts to improve treatment have focused on increasing oral dosages (including the use of high loading doses) and more frequent drug administration regimens (e.g., multiple daily administrations).

[0025] In some embodiments, certain long-acting injectable depot compositions of the present invention have also been found to exhibit high drug loading capacities, which provide many advantages, including the potential use of smaller injection volumes to administer the compositions.

[0026] The present invention fulfills the need for novel treatments for HSV infection that may offer improvements in efficacy and / or safety and / or patient use.

[0027] In a second aspect, the present invention provides a method for forming a pharmaceutical composition according to the first aspect. While various techniques can be used to form the injectable depot compositions of the present invention, certain processes have been found to offer particular advantages. Furthermore, it has also been found that certain chemical and / or physical forms of helicase-primase inhibitors (e.g., forms with low water solubility and low dissolution rates) are well suited for formulation into the injectable depot compositions of the present invention.

[0028] In a third aspect, the present invention provides a method for treating or preventing a herpes infection in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, wherein the helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, is formulated as a pharmaceutical composition according to the first aspect. [Brief explanation of the drawings]

[0029] This Summary, like the Detailed Description below, will be better understood when read in conjunction with the accompanying drawings, in which: For the purpose of illustrating the compositions and methods of the present disclosure, the drawings depict exemplary embodiments of the compositions and methods, but the compositions and methods are not limited to the particular embodiments of the present disclosure.

[0030] [Figure 1] FIG. 1 shows the XRPD diffractogram of plitelivir free base dihydrate.

[0031] [Figure 2]Figure 2 shows the mean plasma concentration-time profiles of pritelivir following subcutaneous administration of the aqueous suspension formulation disclosed in Example 1 at 33.3 mg / kg (open triangles) and 166.7 mg / kg (open squares) in male Sprague Dawley rats (n=3 / arm).

[0032] [Figure 3] FIG. 3 shows the mean plasma concentration-time profile of plitelivir following subcutaneous administration of the aqueous suspension formulation disclosed in Example 2 at 10 mg / kg in male Sprague Dawley rats (n=3).

[0033] [Figure 4] FIG. 4 shows the mean plasma concentration-time profile of plitelivir following subcutaneous administration of the aqueous suspension formulation disclosed in Example 3 at 10 mg / kg in male cynomolgus monkeys (n=3).

[0034] [Figure 5] FIG. 5 shows the mean plasma concentration-time profile of plitelivir following subcutaneous administration of the formulation disclosed in Example 4 at 10 mg / kg in male non-naive beagle dogs (n=3).

[0035] [Figure 6] Figure 6 shows the mean plasma concentration-time profile of pritelivir after intravenous (IV) administration of 1 mg / kg and subcutaneous (SC) administration of 5 mg / kg of the solution disclosed in Example 5 in male Sprague Dawley rats (n=3 / arm).

[0036] [Figure 7] FIG. 7 shows the mean plasma concentration-time profile of Example 7 Compound 1 in male Sprague Dawley rats (n=3) after subcutaneous administration of the aqueous suspension formulation disclosed in Example 7 at 10 mg / kg (triangles).

[0037] [Figure 8] FIG. 8 shows the mean plasma concentration-time profile of Example 7 Compound 1 in male cynomolgus monkeys (n=3) following subcutaneous administration of the aqueous suspension formulation disclosed in Example 7 at 10 mg / kg (triangles).

[0038] [Figure 9] FIG. 9 shows the mean plasma concentration-time profile of Example 7 Compound 1 following subcutaneous administration of the aqueous suspension formulation disclosed in Example 7 at 10 mg / kg (triangles) in male non-naive beagle dogs (n=3).

[0039] [Figure 10] FIG. 10 shows the mean plasma concentration-time profile of Compound 1 (Form A) following subcutaneous administration of the aqueous suspension formulation disclosed in Example 10 at 10 mg / kg (triangles) in male Sprague Dawley rats (n=3).

[0040] [Figure 11] FIG. 11 shows the mean plasma concentration-time profile of Compound 1 (Form A) following subcutaneous administration of the aqueous suspension formulation disclosed in Example 11 at 10 mg / kg (triangles) in male cynomolgus monkeys (n=3).

[0041] [Figure 12] FIG. 12 shows the mean plasma concentration-time profile of Compound 1 (Form A) following subcutaneous administration of the aqueous suspension formulation disclosed in Example 12 at 10 mg / kg (diamonds) in male non-naive beagle dogs (n=3).

[0042] [Figure 13] FIG. 13 shows the mean plasma concentration-time profile of Compound 1 in male Sprague Dawley rats (n=3) following intravenous administration of the solution disclosed in Example 13 at 1 mg / kg (open triangles).

[0043] [Figure 14]FIG. 14 shows the mean plasma concentration-time profile of Compound 1 following intravenous administration of the solution disclosed in Example 13 at 0.25 mg / kg (open triangles) in male cynomolgus monkeys (n=3).

[0044] [Figure 15] FIG. 15 shows the mean plasma concentration-time profile of Compound 1 following intravenous administration of the solution disclosed in Example 13 at 0.15 mg / kg (open triangles) in male non-naive beagle dogs (n=3).

[0045] [Figure 16] FIG. 16 shows the XRPD diffractogram of the sesquihydrate of plitelivir free base.

[0046] [Figure 17] FIG. 17 shows the XRPD diffractogram of anhydrous Form A of Compound 1 free form.

[0047] [Figure 18] FIG. 18 shows the XRPD diffractogram of anhydrous Form C of Compound 1 free form.

[0048] [Figure 19] FIG. 19 shows the mean plasma concentration-time profiles of Compound 1 following IV administration of the solution disclosed in Example 13A to (A) male Sprague Dawley rats (n=3) at 0.2 mg / kg, (B) male non-naive beagle dogs (n=3) at 0.15 mg / kg, (C) male non-naive cynomolgus monkeys (n=3) at 0.2 mg / kg, and (D) naive Bama minipigs (n=3) at 0.25 mg / kg.

[0049] [Figure 20] FIG. 20 shows an SEM image of the microsuspension described in Example 14.

[0050] [Figure 21] FIG. 21 shows an SEM image of the nanosuspension described in Example 14.

[0051] [Figure 22] Figure 22 shows the mean plasma concentration-time profiles of plitelivir following subcutaneous administration of 10 mg / kg of the solution (filled circles), microsuspension (filled triangles), and nanosuspension (filled squares) disclosed in Example 14 in male non-naive beagle dogs (n=3).

[0052] [Figure 23] FIG. 23 shows the mean plasma concentration-time profile of Compound 1 following subcutaneous administration of the solution formulation disclosed in Example 15 (filled triangles) at 100 mg / kg in male Sprague Dawley rats (n=3).

[0053] [Figure 24] FIG. 24 shows the mean plasma concentration-time profiles of Compound 1 following subcutaneous administration of the solution disclosed in Example 16 at 10 mg / kg; 0.1 ml / kg (filled triangles) and 10 mg / kg; 0.2 ml / kg (filled diamonds) in male non-naive beagle dogs (n=3).

[0054] [Figure 25] FIG. 25 shows an SEM image of the microsuspension described in Example 17.

[0055] [Figure 26] FIG. 26 shows an SEM image of the nanosuspension described in Example 17.

[0056] [Figure 27] FIG. 27 shows the mean plasma concentration-time profiles of Compound 1 (Form C) following subcutaneous administration of the microsuspension (filled squares) and nanosuspension (filled triangles) disclosed in Example 17 at 20 mg / rat (approximately 80 mg / kg) in male Sprague Dawley rats (n=3).

[0057] [Figure 28] FIG. 28 shows the mean plasma concentration-time profiles of Compound 1 (Form C) in male Sprague Dawley rats (n=3) following intramuscular administration of the microsuspension (filled squares) and nanosuspension (filled triangles) disclosed in Example 17 at 20 mg / rat (approximately 80 mg / kg). DETAILED DESCRIPTION OF THE INVENTION

[0058] Detailed Description of the Invention The compositions, uses, and methods of the present disclosure may be more readily understood by reference to the following detailed description taken in conjunction with the accompanying drawings, which form a part of this disclosure: It is to be understood that the disclosed compositions and methods are not limited to the specific compositions and methods described and / or illustrated herein, and that the terminology used herein is for the purpose of describing particular embodiments by way of example only, and is not intended to limit the compositions and methods described in the claims.

[0059] definition Unless otherwise stated, the following terms used in the specification and claims have the meanings set forth below.

[0060] A reference to a particular value includes at least that particular value, unless the context clearly dictates otherwise. When a range of values ​​is expressed, another embodiment includes from the one particular value and / or to the other particular value. Also, reference to values ​​stated in a range includes each and every value within that range. All ranges are inclusive and combinable.

[0061] It should be understood that certain features of the compositions and methods of the present disclosure, which are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the compositions and methods of the present disclosure, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any subcombination.

[0062] As used herein, the singular forms "a", "an" and "the" include the plural.

[0063] As used herein, C max means the geometric mean maximum concentration of an active agent, which can be measured in vivo after administering a composition of the invention to a subject and measuring the plasma levels of the drug at various times after administration.

[0064] As used herein, AUC means area under the curve, which is the definite integral of the plasma active drug concentration as a function of time.

[0065] As used herein, a "patient" or "subject" refers to a mammal, including domestic animals, livestock animals, and zoo animals. Conveniently, a "patient" or "subject" is a human being.

[0066] It should be understood that references to "treating" or "treatment" include prophylaxis as well as the alleviation of established symptoms of a condition. Thus, "treating" or "treatment" of a condition, disorder, or condition includes: (1) preventing or delaying the appearance of clinical symptoms of an existing condition, disorder, or condition in a person suffering from or predisposed to the condition, disorder, or condition, but who has not yet experienced or manifested clinical or asymptomatic symptoms of the condition, disorder, or condition; (2) inhibiting the condition, disorder, or condition, i.e., arresting, alleviating, or delaying the onset of the disease or its recurrence (in the case of maintenance therapy), or at least one clinical or asymptomatic symptom thereof; or (3) palliating or attenuating the disease, i.e., causing regression of the condition, disorder, or condition, or at least one clinical or asymptomatic symptom thereof. As used herein, "treating" and similar terms can specifically include reducing the severity and / or frequency of HSV-induced symptoms, eliminating HSV-induced symptoms and / or the underlying causes of such symptoms, reducing the frequency or likelihood of HSV-induced symptoms and / or their underlying causes, delaying, preventing, and / or slowing the progression of HSV-induced pathologies, and ameliorating or repairing damage caused directly or indirectly by HSV infection. The term "preventing," as used herein with respect to HSV infection or an HSV-related disorder, refers to reducing the likelihood of HSV infection. In one embodiment, "treating" a condition, disorder, or condition or "treatment" of a condition, disorder, or condition means (1) inhibiting the condition, disorder, or condition, i.e., preventing, alleviating, or delaying the onset of the disease or its recurrence (in the case of maintenance therapy), or at least one clinical or asymptomatic symptom thereof, or (2) alleviating or attenuating the disease, i.e., causing regression of the condition, disorder, or condition, or at least one clinical or asymptomatic symptom thereof.Advantageously, "treating" and similar terms mean reducing the severity and / or frequency of HSV-induced symptoms, eliminating HSV-induced symptoms and / or the underlying cause of said symptoms, reducing the frequency or likelihood of HSV-induced symptoms and / or their underlying causes, delaying, preventing and / or slowing the progression of HSV-induced pathologies, and / or ameliorating or repairing damage caused directly or indirectly by HSV infection.

[0067] A "therapeutically effective amount" or "therapeutically effective dose" refers to an amount of a compound that, when administered to a patient or subject for treating a disease, is sufficient to effect such treatment of the disease. As used herein, the phrase "therapeutically effective dose" or "therapeutically effective amount" may refer to an amount of a helicase-primase inhibitor administered to a patient or subject using a long-acting injectable pharmaceutical composition as described herein that is effective to achieve a particular biological or therapeutic result, such as, but not limited to, the biological or therapeutic results disclosed, described, or exemplified herein. A therapeutically effective dose may vary depending on factors such as the individual's disease state, age, sex, and weight, as well as the ability of the composition to elicit a desired response in a subject. Such results may include, but are not limited to, relief, remission, and / or regression of a condition caused by or associated with HSV, or prevention of the onset of a condition caused by or associated with HSV, as determined by any means suitable in the art.

[0068] When values ​​are expressed as approximations, by use of the antecedent "about," it is understood that the particular value forms another embodiment. Further, the term "about" refers to a ±10% variation from the nominal value, unless otherwise indicated or inferred.

[0069] The term "about," when used in reference to a numerical range, cutoff value, or specific value, is used to indicate that the stated value may vary by up to 10% from the stated value. Because many of the numerical values ​​used herein were determined experimentally, those skilled in the art should understand that such determinations can, and often do, vary from experiment to experiment. Values ​​used herein should not be considered unduly limited by this inherent variation. Thus, the term "about" is used to include variations of ±10% or less, ±5% or less, ±1% or less, ±0.5% or less, or ±0.1% or less from the stated value.

[0070] At various places in this specification, values ​​are disclosed in groups or ranges, and the description is specifically intended to include all individual subcombinations of the members of such groups and ranges, as well as any combination of the various endpoints of such groups or ranges. For example, integers in the range of 0 to 40 are specifically intended to individually disclose 0, 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, and 40; integers in the range of 1 to 20 are specifically intended to individually disclose 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20.

[0071] As used herein, when a composition is described as having, including, or comprising particular ingredients, or a process is described as having, including, or comprising particular process steps, it is contemplated that a composition of the present teachings also consists essentially of or consists of the recited ingredients, and that a process of the present teachings also consists essentially of or consists of the recited process steps.

[0072] The use of any examples or exemplary language herein, such as, "such as," "including," or "for example," is intended merely to better describe the present teachings and does not limit the scope of the invention unless recited in the claims. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the present teachings.

[0073] Helicase-primase inhibitors The term "helicase-primase inhibitor" in the context of the present invention refers to a compound or agent capable of reducing viral replication by inhibiting the viral complex consisting of DNA helicase, DNA primase, and cofactor subunits. The helicase-primase complex is utilized by herpesviruses, and thus, helicase-primase inhibitors have antiviral activity against one or more herpesviruses, such as herpes simplex virus subtypes 1 and 2 (HSV-1, HSV-2), varicella-zoster virus (VZV), Epstein-Barr virus (EBV), cytomegalovirus (CMV), and one or more human herpesviruses 6-8 (HHV6-8).

[0074] In one embodiment, the helicase-primase inhibitor has antiviral activity against HSV-1 and / or HSV-2.

[0075] In one embodiment, the helicase-primase inhibitor has an in vitro EC50 activity against HSV-1 and / or HSV-2 of less than about 0.1 μM 50 Advantageously, the helicase-primase inhibitor has an in vitro EC value of less than about 0.075 μM or less than about 0.05 μM against HSV-1 and / or HSV-2, advantageously against HSV-1 and HSV-2. 50 More advantageously, the helicase-primase inhibitor has an in vitro EC value of less than about 0.030 μM against HSV-1 and / or HSV-2.50 More advantageously, the helicase-primase inhibitor has an in vitro EC value of less than about 0.020 μM against HSV-1 and / or HSV-2. 50 In vitro EC against HSV-1 and / or HSV-2 50 The value can be determined according to methods known to those skilled in the art, such as those disclosed in Field et al. (2013, Antiviral Res. 100, pp. 297-299). In vitro EC 50 The value may also be determined according to the assays described in the Examples section of this application.

[0076] Helicase-primase inhibitors may undergo plasma protein binding in vivo. In one embodiment, the EC 50 Values ​​represent the unbound plasma concentration of active drug.

[0077] In one embodiment, the helicase-primase inhibitor exhibits no or low levels of carbonic anhydrase inhibition, such as inhibition of carbonic anhydrase I and / or carbonic anhydrase II. Carbonic anhydrase inhibition can be measured using a carbonic anhydrase I assay such as that described by Katritzky et al. (J. Med. Chem. 1987, 30:2058) and a carbonic anhydrase I assay such as that described by Iyer et al. (J. Biomol. Screen 2006, 11:782). In one embodiment, the helicase-primase inhibitor exhibits an IC of greater than 2.0 μM, greater than 3.0 μM, and most preferably greater than 5.0 μM, when measured by either or both of these assays. 50 It has.

[0078] The helicase-primase inhibitor used in the compositions of the present invention has a human terminal biological half-life of 10 hours or more. The human terminal biological half-life of the helicase-primase inhibitor can be derived from the terminal elimination phase of the pharmacokinetic profile after intravenous administration of the helicase-primase inhibitor. The human terminal biological half-life of pritelivir is up to 80 hours (Wald et al., (2014) New England Journal of Medicine 370, pp. 201-210). In one embodiment, the human terminal biological half-life of the helicase-primase inhibitor is 20 hours or more, e.g., 30 hours or more, 40 hours or more, 50 hours or more, 60 hours or more, 70 hours or more, or 80 hours or more. Advantageously, the human terminal biological half-life of the helicase-primase inhibitor is 30 hours or more.

[0079] In one embodiment, the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof has low aqueous solubility. Advantageously, the compound has a solubility in water or an aqueous solvent system (measured at room temperature) of less than 100 μg / ml, less than 50 μg / ml, preferably less than 25 μg / ml, and more preferably less than 20 μg / ml. Advantageously, the compound has a solubility in water at about pH 7.0 (measured at room temperature) of less than 100 μg / ml, less than 50 μg / ml, preferably less than 25 μg / ml, and more preferably less than 20 μg / ml.

[0080] Examples of helicase-primase inhibitors include: · N-methyl-N-(4-methyl-5-sulfamoyl-1,3-thiazol-2-yl)-2-[4-(pyridin-2-yl)phenyl]acetamide (Pritelivir; WO 01 / 47904); (S)-2-(2',5'-difluoro-[1,1'-biphenyl]-4-yl)-N-methyl-N-(4-methyl-5-(S-methylsulfone-imidoyl)thiazol-2-yl)acetamide (IM-250; WO 2017 / 174640 and WO 2019 / 068817 A1); and N-[2-[4-(2-aminothiazol-4-yl)anilino]-2-oxo-ethyl]-N-[(1S)-1-phenylethyl]pyridine-4-carboxamide (BILS-179BS; WO 97 / 24343) Examples include:

[0081] The helicase-primase inhibitors of the present invention include both the parent compound and pharmaceutically acceptable salts of the parent compound.

[0082] In cases where the helicase-primase inhibitor is sufficiently basic or acidic to form a stable pharmaceutically acceptable acid or base salt, preparation and administration of the helicase-primase inhibitor as a pharmaceutically acceptable salt may be appropriate.

[0083] Pharmaceutically acceptable salts can be obtained using standard procedures well known in the art, for example, by reacting a sufficiently basic compound, such as an amine, with a suitable acid that provides a physiologically acceptable anion. Alkali metal (e.g., sodium, potassium, or lithium) or alkaline earth metal (e.g., calcium) salts of carboxylic acids can also be prepared.

[0084] Examples of pharmaceutically acceptable salts are inorganic and organic acid addition salts formed with acids that form physiologically acceptable anions, such as malate, oxalate, chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, superphosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)) salts.

[0085] Helicase-primase inhibitors that are acidic in nature can form base salts with various pharmacologically acceptable cations, including alkali metal or alkaline earth metal salts, particularly calcium, magnesium, sodium, lithium, zinc, potassium, and iron salts.

[0086] It is also to be understood that certain inhibitors of the present invention may exhibit polymorphism, and that the present invention encompasses all such forms (including anhydrous / unsolvated forms, solvates and hydrates).

[0087] In one embodiment, the helicase-primase inhibitor is present in the composition as a crystalline solid. A crystalline solid form is a solid material in which the components of the solid material are arranged in a highly ordered microstructure, thereby forming a crystal lattice that extends in all directions. The crystalline solid may exist in any suitable form, such as an anhydrous form, a hydrated form, or a solvated form. It should also be understood that the helicase-primase inhibitor of the present invention may be pure, essentially pure, or have a purity level with respect to other crystalline forms of greater than 75%, e.g., greater than 85%, greater than 90%, greater than 95%, greater than 97%, greater than 98%, or greater than 99% (e.g., as measured by powder X-ray diffraction).

[0088] Certain crystalline solid forms of helicase-primase inhibitors, such as certain crystalline solid forms of plitelivir, have surprisingly been found to be well suited for the long-acting injectable depot compositions of the present invention.

[0089] In one embodiment, the helicase-primase inhibitor is pritelivir or a pharmaceutically acceptable salt thereof. Pritelivir is N-methyl-N-(4-methyl-5-sulfamoyl-1,3-thiazol-2-yl)-2-<-4-(pyridin-2-yl)phenyl]acetamide, which has the following structure: [ka] N-methyl-N-(4-methyl-5-sulfamoyl-1,3-thiazol-2-yl)-2-[4-(pyridin-2-yl)phenyl]acetamide having the formula:

[0090] Pritelavir has potent antiviral activity against HSV-1 and HSV-2, and in vitro EC 50 The values ​​are understood to be 0.026 μM and 0.029 μM, respectively (Field et al., (2013) Antiviral Res. 100, p. 297-299).

[0091] In one embodiment, the helicase-primase inhibitor is plitelivir free base. The free base of plitelivir has been rejected due to its poor aqueous solubility and its unsuitability for providing a long-term stable formulation (WO 2013 / 045491). However, the applicants have found that the free base compound is particularly suitable for incorporation into the long-acting injectable depot compositions of the present invention.

[0092] In a preferred embodiment, the helicase-primase inhibitor is plitelivir free base, and the free base is present in the composition as a crystalline solid. In one embodiment, the plitelivir crystalline free base is in anhydrous form (anhydrate) or in hydrated form. Advantageously, the plitelivir crystalline free base is a hydrate. Advantageously, the plitelivir crystalline free base is a hemihydrate, dihydrate, or sesquihydrate. More advantageously, the plitelivir crystalline free base is a dihydrate or sesquihydrate. Most advantageously, the plitelivir crystalline free base is a sesquihydrate, as disclosed in the Examples.

[0093] In one embodiment, the helicase-primase inhibitor is [ka] or a pharmaceutically acceptable salt thereof.

[0094] The compound shown above is 2-(3-(2',5'-difluoro-[1,1'-biphenyl]-4-yl)-2-oxotetrahydropyrimidin-1(2H)-yl)-4-methylthiazole-5-sulfonamide (also referred to herein as "Compound 1"), a potent helicase-primase inhibitor.

[0095] Compound 1 inhibited in vitro EC 50 The value was approximately 0.019 μM, and the in vitro EC 50 The in vitro EC value against HSV-1 and / or HSV-2 is 0.011 μM. 50 The value can be determined according to methods known to those skilled in the art, such as those disclosed in Field et al. (2013, Antiviral Res. 100, pp. 297-299). In vitro EC 50 Values ​​may also be determined according to the assays described in the Examples section of this application.

[0096] Compound 1 is predicted to have a predicted human biological terminal half-life of greater than 180 hours (see Example 13 and Example 13A).

[0097] Compound 1 exhibits no or low levels of carbonic anhydrase inhibition, such as inhibition of carbonic anhydrase I and / or carbonic anhydrase II. Carbonic anhydrase inhibition can be measured using a carbonic anhydrase I assay such as that described by Katritzky et al. (J. Med. Chem. 1987, 30:2058) and Iyer et al. (J. Biomol. Screen 2006, 11:782).

[0098] It is to be understood that Compound 1 may exhibit polymorphism, and that the present invention encompasses all such forms, including anhydrous / unsolvated forms, solvates and hydrates.

[0099] In one embodiment, Compound 1 is present in the composition as a crystalline solid. The crystalline solid can be present in any suitable form, such as an anhydrous, hydrated, or solvated form. It should also be understood that Compound 1 can be pure, essentially pure, or have a purity level of greater than 75%, e.g., greater than 85%, greater than 90%, greater than 95%, greater than 97%, greater than 98%, or greater than 99% with respect to other crystalline forms (e.g., as measured by X-ray powder diffraction).

[0100] In a preferred embodiment, the helicase-primase inhibitor present in the composition of the present invention is Compound 1 free form, which is present in the composition as a crystalline solid. In a preferred embodiment, the Compound 1 crystalline free form is an anhydrous form. It has been surprisingly found that certain crystalline solid forms of the helicase-primase inhibitor Compound 1 are well suited for the long-acting injectable depot compositions of the present invention. Conveniently, the Compound 1 crystalline free form is Form A or Form C, as described in the Examples. More conveniently, the Compound 1 crystalline free form is Form C, as described in the Examples.

[0101] Pharmaceutical compositions of the present invention In a first aspect, the present invention provides a long-acting injectable depot pharmaceutical composition comprising a helicase-primase inhibitor or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients, wherein the helicase-primase inhibitor has a human biological terminal half-life of 10 hours or more.

[0102] As used herein, "long-acting injectable depot pharmaceutical composition" refers to a pharmaceutical composition that is adapted for administration to a subject (preferably a human) via a subcutaneous or intramuscular route, and that upon administration forms a depot from which the active ingredient of the composition (a helicase-primase inhibitor) is released over an extended period of time.

[0103] In one embodiment, extended release refers to continuous release of the active ingredient for at least 10 days, e.g., at least 14 days, at least 21 days, at least 28 days, at least 56 days, at least 72 days, at least 96 days, at least 120 days, at least 180 days, at least 240 days, or at least 300 days. Extended release of the active ingredient means that the active ingredient plasma level can be maintained at a therapeutically effective plasma concentration for at least 10 days, e.g., at least 14 days, at least 21 days, at least 28 days, at least 56 days, at least 72 days, at least 96 days, at least 120 days, at least 180 days, at least 240 days, or at least 300 days after administration of the composition, which one of ordinary skill in the art would understand to correspond to "long-acting."

[0104] It should be understood that the release rate of the helicase-primase inhibitor may vary during the extended-release period, e.g., a short "initial burst" of active agent may be observed immediately after administration, followed by a period of lower release. However, extended-release means that the plasma level of the helicase-primase inhibitor may be maintained at a therapeutically effective plasma concentration throughout most, if not all, of the extended-release period.

[0105] In one embodiment, the pharmaceutical composition provides a short "initial burst" of active agent immediately after administration, followed by a period of lower release. Advantageously, Applicants have discovered that certain compositions of the present invention can provide a rapid initial release to achieve high plasma levels of a helicase-primase inhibitor, followed by a continuous release to maintain plasma levels at therapeutically effective concentrations over an extended period of time.

[0106] In one embodiment, the pharmaceutical composition releases less than 20% by weight (conveniently, less than 10%) of the total amount of active agent contained within the pharmaceutical composition within a 24 hour (conveniently, 48 hour) period after administration.

[0107] Advantageously, applicants have discovered that the extended release provided by certain compositions of the present invention, and the resulting sustained plasma levels at therapeutically effective concentrations over an extended period of time, allows for the potential use of lower doses of the helicase-primase inhibitor than would be expected. In one embodiment, the dose administered in a single administration of a long-acting injectable depot pharmaceutical composition capable of providing extended release for at least 14 days (conveniently, at least 28 days, at least 56 days, at least 96 days, at least 120 days, at least 180 days, at least 240 days, or at least 300 days) is about 100-1200 mg (conveniently, 300-900 mg) of the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof.

[0108] In one embodiment, the dose administered in a single administration of the long-acting injectable depot pharmaceutical composition capable of providing extended release over a period of at least one month is about 100-900 mg (conveniently, 100-600 mg) of the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof.

[0109] In one embodiment, the dose administered in a single administration of the long-acting injectable depot pharmaceutical composition capable of providing extended release over a period of at least two months is about 200-900 mg (conveniently, 200-600 mg) of the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof.

[0110] In one embodiment, the dose administered in a single administration of the long-acting injectable depot pharmaceutical composition capable of providing extended release over a period of at least three months is about 300-900 mg (conveniently, 300-600 mg) of the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof.

[0111] In one embodiment, the dose administered in a single administration of the long-acting injectable depot pharmaceutical composition capable of providing extended release over a period of at least six months is about 300-1200 mg (conveniently, 300-900 mg) of the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof.

[0112] In some embodiments, certain long-acting injectable depot compositions of the present invention have been found to exhibit high drug loading capacities, which provide a number of advantages, including the ability to administer the compositions in small injection volumes. In one embodiment, a dose is administered in a single administration of the long-acting injectable depot pharmaceutical composition in an injection volume of about 0.1 to about 5 mL. Conveniently, the dose is administered in an injection volume of about 0.1 to about 3 mL, about 0.1 to about 1.5 mL, or about 0.1 to about 0.75 mL. More conveniently, the dose is administered in an injection volume of about 0.1 to about 0.5 mL.

[0113] In a preferred embodiment, the pharmaceutical composition is designed for subcutaneous or intramuscular injection. In a further aspect of the invention, the pharmaceutical composition is for subcutaneous administration. In yet another aspect of the invention, the pharmaceutical composition is for intramuscular administration. Advantageously, the dose of the helicase-primase inhibitor is administered subcutaneously in an injection volume of about 0.1 to about 3 mL, or about 0.1 to about 1.5 mL. Advantageously, the dose of the helicase-primase inhibitor is administered intramuscularly in an injection volume of about 0.1 to about 5 mL, or about 0.1 to about 3 mL.

[0114] The advantages of the present invention are not limited to a particular type of long-acting injectable depot pharmaceutical composition having a particular drug release mechanism. Pharmaceutical compositions suitable for subcutaneous and / or intramuscular administration can be aqueous-based depots, aqueous-based suspension depots, organic solvent-based solution formulations, organic solvent-based suspension depots, gel-based depots, in-situ gelling depot formulations, oil-based depots, emulsion-based depots, monolithic polymer-based depots, microparticle polymer-based depots, or solid implant depots.

[0115] In one embodiment, the pharmaceutical composition of the present invention includes a rate-controlling agent that facilitates the extended release of the helicase-primase inhibitor. In one embodiment, the rate-controlling agent includes at least one biodegradable polymer. Suitable biodegradable polymers are those that are degradable under physiological conditions so that the polymer and its degradation products do not elicit unacceptable toxicity or immune responses. Such polymers may be of natural or synthetic origin. Examples of natural biodegradable polymers include polysaccharides such as chitosan, alginate, and dextran, or polyesters such as polyhydroxyalkanoates. Examples of synthetic biodegradable polymers include polymers of lactic acid and glycolic acid, as well as copolymers thereof. In one embodiment, the biodegradable polymer is poly(lactic-co-glycolic acid) or polylactic acid, each end-capped with either acid or ester groups. Advantageously, the biodegradable polymer is poly(lactic-co-glycolic acid). Poly(lactic-co-glycolic acid) is also referred to herein as PLGA. Advantageously, the biodegradable polymer is PLGA end-capped with acid groups, hi one embodiment, the biodegradable polymer is PLGA, which has a weight content of about 5-95% lactic acid, the remainder being glycolic acid.

[0116] In advantageous embodiments, the pharmaceutical compositions of the present invention are aqueous based suspensions, organic solvent solution formulations, or in-situ gelling depot formulations.

[0117] In a preferred embodiment, the pharmaceutical composition of the present invention is an aqueous suspension depot. Surprisingly, it has been found that certain aqueous suspension depot compositions provide therapeutically effective plasma levels of the active ingredient for extended periods of time without the need for rate-controlling agents. This can be particularly advantageous because it allows for high drug loading, smaller administration volumes, and lower excipient levels. In one embodiment, the pharmaceutical composition of the present invention does not contain a rate-controlling agent or contains a rate-controlling agent in an amount of less than 20% by weight, or less than 15%, 10%, 5%, 3%, 2%, or 1% by weight of the total composition. Advantageously, the pharmaceutical composition of the present invention does not contain a rate-controlling agent, such as a biodegradable polymer.

[0118] It has also been found that the particle size of the helicase-primase inhibitor in the aqueous suspension depot composition influences release. Suitable methods for measuring the particle size distribution of a sample will be apparent to those skilled in the art and include techniques such as laser diffraction. Dx (or Dv x The term D50 (or Dv) refers to the particle size up to and including the particle size at which x% of the total volume of material in the sample is contained. 50 ) is 10 μm, it means that 50% of the sample has a particle size of 10 μm or less.

[0119] In one embodiment, the particle size volume distribution of suspended particles of the active agent (helicase-primase inhibitor) in the aqueous suspension depot composition is such that less than 10% of the particles have a size greater than 9 microns, as determined by laser diffraction analysis. In one embodiment, the aqueous suspension depot composition comprises particles of the active agent having a D10 of 0.5-2.5 μm, a D50 of 2-5 μm, and / or a D90 of 4-13 μm. In one embodiment, the aqueous suspension depot composition comprises particles of the active agent having a D10 of 0.5-2.5 μm, a D50 of 3-5 μm, and / or a D90 of 8-13 μm. In one embodiment, the aqueous suspension depot composition comprises particles of the active agent having a D10 of 0.5-2.0 μm, a D50 of 2-3 μm, and / or a D90 of 4-8 μm. In one embodiment, the aqueous suspension depot composition comprises particles of active agent having a D10 of 1-2 μm, a D50 of 4-5 μm, and / or a D90 of 10-11 μm. In one embodiment, the aqueous suspension depot composition comprises particles of active agent having a D90 of 10-5000 nm, for example 20-2000 nm, or 50-1000 nm. Conveniently, the helicase inhibitor is plitelivir.

[0120] In one embodiment, the helicase-primase inhibitor is Compound 1, and the aqueous suspension depot composition comprises particles of the active agent having a D10 of 0.3-1.5 μm, a D50 of 2-4 μm, and / or a D90 of 8-16 μm. In one embodiment, the aqueous suspension depot composition comprises particles of the active agent having a D10 of 0.3-0.7 μm, a D50 of 2-3 μm, and / or a D90 of 12-16 μm.

[0121] In one embodiment, the helicase-primase inhibitor is pritelivir, and the pharmaceutical composition is an aqueous-based microsuspension depot composition. Conveniently, the particle size volume distribution of suspended particles of pritelivir in the aqueous-based microsuspension depot composition is such that less than 10% of the particles have a size greater than 20 μm, as measured by laser diffraction analysis. In one embodiment, the aqueous-based microsuspension depot composition comprises particles of pritelivir having a D90 of 4-20 μm, advantageously 5-15 μm, and more advantageously 6-10 μm. In one embodiment, the aqueous-based microsuspension depot composition comprises particles of pritelivir having a D10 of 0.3-3 μm, a D50 of 2-6 μm, and / or a D90 of 5-15 μm. In one embodiment, the aqueous microsuspension depot composition comprises particles of plitelivir having a D10 of 1.5-2.5 μm, a D50 of 3-5 μm, and / or a D90 of 6-10 μm.

[0122] In one embodiment, the helicase-primase inhibitor is pritelivir, and the pharmaceutical composition is an aqueous nanosuspension depot composition. Advantageously, the particle size volume distribution of suspended particles of pritelivir in the aqueous nanosuspension depot composition is such that less than 10% of the particles have a size greater than 700 nm, as measured by laser diffraction analysis. In one embodiment, the aqueous nanosuspension depot composition comprises particles of pritelivir having a D90 of 1-700 nm, advantageously 1-500 nm, and more advantageously 1-300 nm.

[0123] In one embodiment, the helicase-primase inhibitor is Compound 1, and the pharmaceutical composition is an aqueous microsuspension depot composition. Advantageously, the particle size volume distribution of suspended particles of Compound 1 in the aqueous microsuspension depot composition is such that less than 10% of the particles have a size greater than 15 μm, as measured by laser diffraction analysis. In one embodiment, the aqueous microsuspension depot composition comprises particles of Compound 1 having a D90 of 4-13 μm, advantageously 4-10 μm, and more advantageously 4-8 μm. In one embodiment, the aqueous microsuspension depot composition comprises particles of Compound 1 having a D10 of 0.3-2 μm, a D50 of 2-4 μm, and / or a D90 of 4-13 μm. In one embodiment, the aqueous microsuspension depot composition comprises particles of Compound 1 having a D10 of 0.3-1.5 μm, a D50 of 2-3.5 μm, and / or a D90 of 4-8 μm.

[0124] In one embodiment, the helicase-primase inhibitor is Compound 1, and the pharmaceutical composition is an aqueous nanosuspension depot composition. Advantageously, the particle size volume distribution of suspended particles of Compound 1 in the aqueous nanosuspension depot composition is such that less than 10% of the particles have a size greater than 500 nm, as measured by laser diffraction analysis. In one embodiment, the aqueous nanosuspension depot composition comprises particles of Compound 1 having a D90 of 1-400 nm, advantageously 1-300 nm, and more advantageously 1-200 nm. In one embodiment, the aqueous nanosuspension depot composition comprises particles of Compound 1 having a D10 of 1-100 nm, a D50 of 1-200 nm, and / or a D90 of 1-300 nm. In one embodiment, the aqueous nanosuspension depot composition comprises particles of Compound 1 having a D10 of 1-100 nm, a D50 of 1-150 nm, and / or a D90 of 1-200 nm. In one embodiment, the aqueous nanosuspension depot composition comprises particles of Compound 1 having a D10, D50 and D90 of 1 to 100 nm.

[0125] In one embodiment, the pharmaceutical composition of the present invention is a solution formulation in an organic solvent. The organic solvent can be any suitable organic solvent that solubilizes the helicase-primase inhibitor and is biocompatible. A suitable biocompatible organic solvent diffuses out of the formulation and solidifies it after injection into the body. Advantageously, the biocompatible solvent is non-toxic, water-miscible, and does not cause severe tissue irritation or necrosis at the injection / implantation site. Examples of such solvents include benzyl alcohol, benzyl benzoate, N-methyl-2-pyrrolidone, 2-pyrrolidone, ethanol, propylene glycol, acetone, methyl acetate, ethyl acetate, methyl ethyl ketone, dimethylacetamide, dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, caprolactam, oleic acid, and 1-dodecylazacycloheptan-2-one. In one embodiment, the organic solvent is selected from N-methylpyrrolidone (NMP), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), benzyl alcohol, benzyl benzoate, and mixtures thereof. The organic solvent solution formulation may further comprise an aqueous medium (such as an aqueous buffer or saline) and / or a surfactant (such as a polyethylene glycol fatty acid ester, e.g., PEG-15 hydroxystearate).

[0126] In one embodiment, the helicase-primase inhibitor is Compound 1, and the pharmaceutical composition is an organic solvent solution formulation. In one embodiment, the organic solvent solution formulation comprises Compound 1 dissolved in an organic solvent selected from N-methylpyrrolidone (NMP), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), benzyl alcohol, benzyl benzoate, and mixtures thereof, preferably N-methylpyrrolidone (NMP), dimethylacetamide (DMA), and dimethyl sulfoxide (DMSO), e.g., NMP. In one embodiment, the organic solvent solution formulation comprising Compound 1 may optionally further comprise an aqueous medium (e.g., aqueous buffer or saline) and / or a surfactant (e.g., polyethylene glycol fatty acid ester, e.g., PEG-15 hydroxystearate). In one embodiment, the composition comprises 5 to 95% v / v (e.g., 5 to 75% v / v, 5 to 50% v / v, 5 to 20% v / v, or 5 to 15% v / v) of an organic solvent selected from N-methylpyrrolidone (NMP), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), and mixtures thereof. In one embodiment, the composition comprises Compound 1 at a concentration of 1 to 500 mg / mL, e.g., 10 to 400 mg / mL, 100 to 400 mg / mL, or 200 to 300 mg / mL. In one embodiment, the composition comprises Compound 1 at a concentration of 100 to 400 mg / mL, and 5 to 95% v / v of NMP as the organic solvent.

[0127] In one embodiment, the helicase-primase inhibitor is pritelivir free base, and the pharmaceutical composition is an organic solvent solution formulation. In one embodiment, the organic solvent solution formulation comprises Compound 1 dissolved in an organic solvent selected from N-methylpyrrolidone (NMP), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), benzyl alcohol, benzyl benzoate, and mixtures thereof, preferably N-methylpyrrolidone (NMP), dimethylacetamide (DMA), and dimethyl sulfoxide (DMSO). In one embodiment, the organic solvent solution formulation containing pritelivir free base may further comprise an aqueous medium (e.g., aqueous buffer or saline) and / or a surfactant (e.g., polyethylene glycol fatty acid ester, e.g., PEG-15 hydroxystearate). In one embodiment, the composition comprises 5 to 95% v / v (e.g., 5 to 75% v / v, 5 to 50% v / v, 5 to 20% v / v, or 5 to 15% v / v) of an organic solvent selected from N-methylpyrrolidone (NMP), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), and mixtures thereof. In one embodiment, the composition comprises plitelivir free base at a concentration of 1 to 500 mg / mL, e.g., 10 to 400 mg / mL, 50 to 300 mg / mL, or 50 to 200 mg / mL. In one embodiment, the composition comprises plitelivir free base at a concentration of 10 to 400 mg / mL (e.g., 100 to 400 mg / mL) and 5 to 95% v / v of NMP as the organic solvent.

[0128] In one embodiment, the pharmaceutical composition of the present invention is an in situ gelling depot formulation. As used herein, the term "in situ gelling depot formulation" refers to a formulation comprising a helicase-primase inhibitor, a biodegradable polymer (such as the biodegradable polymers described above as rate-controlling agents), and a biocompatible solvent. This formulation is delivered to a patient as an injectable liquid, but solidifies into a solid depot formulation as the liquid solvent diffuses in vivo. A suitable biocompatible solvent is one that can dissolve the components of the formulation and diffuses out of the formulation after injection into the body, solidifying the formulation. Advantageously, the solvent for the biodegradable polymer is nontoxic, water-miscible, and otherwise biocompatible. The solvent must also be biocompatible so as not to cause severe tissue irritation or necrosis at the implantation site. Furthermore, the solvent must be water-miscible so as to rapidly diffuse into body fluids, allowing the polymer solution to penetrate water and solidify or solidify. Examples of such solvents include benzyl alcohol, N-methyl-2-pyrrolidone, 2-pyrrolidone, ethanol, propylene glycol, acetone, methyl acetate, ethyl acetate, methyl ethyl ketone, dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, caprolactam, oleic acid, and 1-dodecylazacycloheptan-2-one.

[0129] The compositions of the present invention include one or more pharmaceutically acceptable excipients. The long-acting injectable depot pharmaceutical compositions of the present invention include at least one excipient selected from the group consisting of solvents, co-solvents, wetting or suspending agents, isotonicity agents, pH adjusters, stabilizers, emulsifiers, and viscosity adjusters.

[0130] Suitable wetting or suspending agents for inclusion in the formulations of the present invention include cellulose derivatives, polyvinylpyrrolidone, polysorbate 20 and polysorbate 80, lecithin, polyoxyethylene sorbitan fatty acid esters, and polyoxyethylene sorbitan fatty acid esters. Advantageously, polysorbate 20 is used alone or in combination. Advantageously, polysorbate 80 is present in the composition. Advantageously, the composition contains about 0.2 wt% to about 5 wt% polysorbate 80, more advantageously about 0.2 wt% to about 2 wt% polysorbate 80.

[0131] The isotonic agent that can be contained in the long-acting injectable depot pharmaceutical composition of the present invention is, for example, sodium chloride, dextrose, mannitol, sorbitol, lactose, and / or sodium sulfate.Conveniently, the isotonic agent is sodium chloride.Conveniently, the isotonic agent is mannitol.

[0132] The pH adjusting agent contained in the long-acting injectable depot pharmaceutical composition of the present invention may be, but is not limited to, hydrochloric acid or sodium hydroxide.

[0133] The depot formulations of the present invention may further contain a preservative, which may be selected from the group consisting of benzoic acid, benzyl alcohol, butylated hydroxyanisole, butylated hydroxytoluene, chlorbutol, gallates, hydroxybenzoates, EDTA, phenol, parabens, methylparaben, propylparaben, butylparaben, benzalkonium chloride, thiomerosal, metacresol, or chlorobutanol.

[0134] In some embodiments, the long-acting injectable depot pharmaceutical compositions of the present invention comprise at least one viscosity modifier. Optionally, the viscosity modifier is selected from the group consisting of sodium carboxymethylcellulose, methylcellulose, hydroxypropylcellulose, calcium carboxymethylcellulose, cross-linked carboxymethylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, acacia, gelatin, and polyvinylpyrrolidone. Advantageously, the composition comprises about 0.1 wt% to about 5 wt% of the viscosity modifier. In some embodiments, the viscosity modifier is selected from the group consisting of sodium carboxymethylcellulose and / or hydroxypropylcellulose and / or polyvinylpyrrolidone.

[0135] The long-acting injectable depot pharmaceutical compositions of the present invention can be stored, for example, by lyophilization in suitable vials and resuspended for injection immediately prior to injection. The compositions can be stored in pre-filled syringes.

[0136] Advantageously, the compositions of the present invention can exhibit good stability during storage. Thus, in one embodiment, the compositions are stable for at least 4 weeks, e.g., at least 8 weeks, or at least 12 weeks. In this context, "stable" can refer to the physical and / or chemical stability of the active agent (helicase-primase inhibitor) within the long-acting injectable depot pharmaceutical composition. "Stable" can also refer to the maintenance of the release profile or pharmacokinetics of the composition over the stated storage period. "Stable" can also refer to the appearance of the composition; for example, in the case of an aqueous suspension depot composition, it can refer to the maintenance of the uniformity of the suspension over the stated storage period.

[0137] Physical stability relates to maintaining the active agent in the same physical form. When the long-acting injectable depot pharmaceutical composition is an aqueous suspension and the active agent is suspended as a crystalline solid, the physical form of the suspended solid (as determined by XRPD) does not change during the stated storage period.

[0138] Chemical stability refers to the low level of impurities formed over the stated storage period. Typically, these are impurities associated with the active drug and can be measured by suitable techniques such as HPLC or LC-MS.

[0139] The release of active agents from the compositions of the present invention can be determined by methods known in the art. For example, the release rate can be determined using in vitro dissolution tests that mimic an aqueous physiological-type environment. As used herein, the term "aqueous physiological-type environment" refers to the body of a warm-blooded animal, particularly the human body. In certain embodiments, the compositions of the present invention are designed for subcutaneous or intramuscular administration, and the term "aqueous physiological-type environment" for such compositions refers to the subcutaneous or intramuscular environment of such a body. These conditions can be simulated in vitro by placing the composition in an aqueous dissolution medium, which may be buffered to physiological pH, at a temperature of 35-40°C. The amount of active agent released over a given period of time can be determined by sampling the dissolution medium and measuring the concentration of the active agent using an appropriate analytical method, such as HPLC.

[0140] The release of an active agent from a composition of the invention can also be determined by in vivo methods known in the art. For example, in vivo release can be tested by measuring the plasma concentration at predetermined time intervals to obtain a plasma concentration versus time profile of the compound of interest.

[0141] Other tests can also be used to measure the amount of active agent released in vivo. Animals (e.g., mice, rats, dogs, etc.) can be used as models to examine release characteristics. For example, in the case of subcutaneous or intramuscular compositions, the composition under investigation can be administered to the animal, and after a specific period of time, the animal can be sacrificed, and the subcutaneous or intramuscular composition can be extracted / collected and analyzed. By measuring the content of active agent remaining in the composition for a specific period of time, the amount and extent of release can be calculated.

[0142] Advantageously, the compositions of the present invention are capable of maintaining therapeutically effective plasma concentrations of the drug for extended periods of time.

[0143] Pharmacokinetics The present invention provides long-acting injectable depot pharmaceutical compositions that offer certain advantages over prior art compositions in that they provide stable and sustained plasma helicase-primase inhibitor levels over extended periods of time, such that viral shedding and viral reactivation may be reduced or even eliminated.

[0144] In one embodiment, a long-acting injectable depot pharmaceutical composition according to the invention maintains a geometric mean plasma concentration of the helicase-primase inhibitor in a subject in need of treatment of at least 25 ng / mL for at least 14 days after administration to the subject.

[0145] In one embodiment, a long-acting injectable depot pharmaceutical composition according to the invention comprises a helicase-primase inhibitor or a pharmaceutically acceptable salt thereof and produces a plasma concentration of the helicase-primase inhibitor of at least 25 ng / mL (e.g., at least 50 ng / mL, or at least 100 ng / mL) in a subject following administration for at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or substantially all) of the dosing interval. Advantageously, the dosing interval is at least 10 days (e.g., at least 14 days, at least 21 days, at least 28 days, at least 56 days, at least 72 days, at least 96 days, at least 120 days, at least 180 days, at least 240 days, or at least 300 days). In one embodiment, the dosing interval is at least 14 days or at least 28 days. Advantageously, the interval between doses is at least 56 days, at least 96 days, at least 120 days, at least 180 days, at least 240 days, or at least 300 days.

[0146] In one embodiment, a long-acting injectable depot pharmaceutical composition according to the invention comprises pritelivir free base and produces a plasma concentration of pritelivir of at least 25 ng / mL (e.g., at least 50 ng / mL, or at least 100 ng / mL) in a subject following administration for at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or substantially all) of the administration interval. Advantageously, the administration interval is at least 10 days (e.g., at least 14 days, at least 21 days, at least 28 days, at least 56 days, at least 72 days, at least 96 days, at least 120 days, at least 180 days, at least 240 days, or at least 300 days). In one embodiment, the administration interval is at least 14 days or at least 28 days. Advantageously, the administration interval is at least 56 days, at least 96 days, at least 120 days, at least 180 days, at least 240 days, or at least 300 days.

[0147] In one embodiment, a long-acting injectable depot pharmaceutical composition according to the invention comprises the free form of Compound 1 and produces a plasma concentration of Compound 1 of at least 25 ng / mL (e.g., at least 50 ng / mL, or at least 100 ng / mL) in a subject after administration for at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or substantially all) of the dosing interval. Advantageously, the dosing interval is at least 10 days (e.g., at least 14 days, at least 21 days, at least 28 days, at least 56 days, at least 72 days, at least 96 days, at least 120 days, at least 180 days, at least 240 days, or at least 300 days). In one embodiment, the dosing interval is at least 14 days or at least 28 days. Advantageously, the dosing interval is at least 56 days, at least 96 days, at least 120 days, at least 180 days, at least 240 days, or at least 300 days.

[0148] Helicase-primase inhibitors typically undergo plasma protein binding. Depending on the degree of plasma protein binding, the free fraction (unbound active drug) may be low compared to the protein-bound fraction. For example, plitelivir is typically 97-98% protein bound, resulting in a free fraction of only 2-3% of the total plasma concentration. In one embodiment, the plasma concentrations referred to above refer to unbound plasma concentrations. Thus, in one embodiment, a long-acting injectable depot pharmaceutical composition according to the present invention maintains a geometric mean unbound plasma concentration of the helicase-primase inhibitor (e.g., plitelivir) of at least 25 ng / mL in a subject in need of such treatment for at least 14 days after administration to the subject. In one embodiment, a long-acting injectable depot pharmaceutical composition of the invention maintains a geometric mean total (unbound and protein-bound) plasma concentration of a helicase-primase inhibitor (e.g., plitelivir) of at least 1,500 ng / mL (e.g., at least 2,000 ng / mL, or at least 2,500 ng / mL) for at least 14 days in a subject in need of treatment following administration to the subject.

[0149] In one embodiment, a long-acting injectable depot pharmaceutical composition according to the invention maintains a geometric mean total (unbound and protein-bound) plasma concentration of Compound 1 of at least 500 ng / mL (e.g., at least 1000 ng / mL, at least 1100 ng / mL, at least 1500 ng / mL, at least 2000 ng / mL, or at least 2500 ng / mL) for most, if not all, of at least 14 days in a subject in need of such treatment after administration to the subject. Advantageously, a long-acting injectable depot pharmaceutical composition according to the invention maintains a geometric mean total (unbound and protein-bound) plasma concentration of Compound 1 of between 500 ng / mL and 1700 ng / mL (e.g., between 900 ng / mL and 1,300 ng / mL, between 900 ng / mL and 1,200 ng / mL) for most, if not all, of at least 14 days in a subject in need of such treatment after administration to the subject.

[0150] Specific Compositions of the Invention In one embodiment, the present invention provides a method for producing a pharmaceutical composition comprising: a) helicase-primase inhibitors, and b) one or more pharmaceutically acceptable excipients The present invention provides a long-acting injectable depot pharmaceutical composition comprising a helicase primase inhibitor having a human biological terminal half-life of 10 hours or more and a solubility in water or an aqueous solvent system of less than 100 μg / ml (measured at room temperature). Advantageously, the long-acting injectable depot pharmaceutical composition sustainably releases the helicase primase inhibitor at a rate that results in a therapeutic plasma concentration of the helicase primase inhibitor for at least one month after subcutaneous or intramuscular administration. Advantageously, the long-acting injectable depot pharmaceutical composition comprises about 1 wt% to about 80 wt% of the helicase primase inhibitor. Advantageously, the long-acting injectable depot pharmaceutical composition comprises about 10 wt% to about 40 wt% of the helicase primase inhibitor. Advantageously, the long-acting injectable depot pharmaceutical composition contains 300-900 mg of the helicase-primase inhibitor, and is administered in an injection volume of about 0.1 to about 3 mL. Advantageously, the long-acting injectable depot pharmaceutical composition comprises an aqueous suspension or an organic solvent solution formulation. Advantageously, the aqueous suspension is a microsuspension or nanosuspension. Advantageously, the long-acting injectable depot pharmaceutical composition does not contain a rate-controlling agent, such as a biodegradable polymer, or contains a rate-controlling agent in an amount of less than 20% by weight, or less than 15%, 10%, 5%, 3%, 2%, or 1% by weight of the total composition.

[0151] In one embodiment, the present invention provides a method for producing a pharmaceutical composition comprising: a) pritelivir free base form (conveniently a hydrate, e.g., hemihydrate, sesquihydrate or dihydrate), and b) one or more pharmaceutically acceptable excipients 1. A long-acting injectable depot pharmaceutical composition comprising: the long-acting injectable depot pharmaceutical composition sustainably releases pritelivir at a rate that results in a therapeutic plasma concentration of the helicase-primase inhibitor for at least one month after subcutaneous or intramuscular administration; A long-acting injectable depot pharmaceutical composition is provided.

[0152] In one embodiment, the present invention provides a method for producing a pharmaceutical composition comprising: a) pritelivir free base form (conveniently the dihydrate or trihydrate); and b) one or more pharmaceutically acceptable excipients 1. A long-acting injectable depot pharmaceutical composition comprising: the long-acting injectable depot pharmaceutical composition sustainably releases pritelivir at a rate that results in a therapeutic plasma concentration of the helicase-primase inhibitor for at least one month after subcutaneous or intramuscular administration; A long-acting injectable depot pharmaceutical composition is provided.

[0153] In one embodiment, the present invention provides a method for producing a pharmaceutical composition comprising: a) pritelivir free base form (conveniently a hemihydrate, dihydrate or sesquihydrate, e.g., a sesquihydrate), and b) one or more pharmaceutically acceptable excipients 1. A long-acting injectable depot pharmaceutical composition comprising: the long-acting injectable depot pharmaceutical composition comprises 300 to 900 mg of pritelivir, and the long-acting injectable depot pharmaceutical composition is administered in an injection volume of about 0.1 to about 3 mL. A long-acting injectable depot pharmaceutical composition is provided. Advantageously, the long-acting injectable depot pharmaceutical composition continuously releases plitelivir at a rate that results in therapeutic plasma concentrations of plitelivir for at least three months (preferably at least six months) after subcutaneous or intramuscular administration. Advantageously, the long-acting injectable depot pharmaceutical composition comprises an organic solvent solution formulation containing plitelivir, wherein the plitelivir is dissolved in an organic solvent selected from N-methylpyrrolidone (NMP), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), benzyl alcohol, benzyl benzoate, and mixtures thereof, preferably N-methylpyrrolidone (NMP), dimethylacetamide (DMA), and dimethyl sulfoxide (DMSO). The composition may further comprise an aqueous medium (such as an aqueous buffer or saline) and / or a surfactant (such as a polyethylene glycol fatty acid ester, e.g., PEG-15 hydroxystearate). Advantageously, the long-acting injectable depot pharmaceutical composition comprises an aqueous suspension. Advantageously, the aqueous suspension is a nanosuspension.

[0154] Advantageously, the long-acting injectable depot pharmaceutical composition does not contain a rate-controlling agent, such as a biodegradable polymer, or contains less than 20% by weight of the total composition of the rate-controlling agent, or less than 15%, less than 10%, less than 5%, less than 3%, less than 2%, or less than 1% by weight. Advantageously, the plitelivir free base form is a sesquihydrate.

[0155] In one embodiment, the present invention provides a method for producing a pharmaceutical composition comprising: c) pritelivir free base form (conveniently the dihydrate or sesquihydrate), and d) one or more pharmaceutically acceptable excipients 1. A long-acting injectable depot pharmaceutical composition comprising: the long-acting injectable depot pharmaceutical composition comprises 300 to 900 mg of a helicase-primase inhibitor, the long-acting injectable depot pharmaceutical composition is administered in an injection volume of about 0.1 to about 2 mL, and the long-acting injectable depot pharmaceutical composition comprises an aqueous suspension (e.g., an aqueous-based microsuspension); A long-acting injectable depot pharmaceutical composition is provided. Advantageously, the pritelivir free base form is a sesquihydrate.

[0156] In one embodiment, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. a) Compound 1 free form (conveniently, anhydrous Form A or Form C), and b) one or more pharmaceutically acceptable excipients 1. A long-acting injectable depot pharmaceutical composition comprising: the long-acting injectable depot pharmaceutical composition continuously releases Compound 1 at a rate that results in therapeutic plasma concentrations of Compound 1 for at least one month after subcutaneous or intramuscular administration; A long-acting injectable depot pharmaceutical composition is provided.Conveniently, the long-acting injectable depot pharmaceutical composition is administered by intramuscular injection.

[0157] In one embodiment, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. a) the free form of Compound 1 (conveniently, anhydrous Form A or Form C), and b) one or more pharmaceutically acceptable excipients 1. A long-acting injectable depot pharmaceutical composition comprising: the long-acting injectable depot pharmaceutical composition comprises 300 to 900 mg of Compound 1, and the long-acting injectable depot pharmaceutical composition is administered in an injection volume of about 0.1 to about 3 mL; A long-acting injectable depot pharmaceutical composition is provided. Advantageously, the long-acting injectable depot pharmaceutical composition sustainably releases Compound 1 at a rate that results in therapeutic plasma concentrations of Compound 1 for at least three months (preferably at least six months) after subcutaneous or intramuscular administration. Advantageously, the long-acting injectable depot pharmaceutical composition comprises an organic solvent solution formulation containing Compound 1, wherein Compound 1 is dissolved in an organic solvent selected from N-methylpyrrolidone (NMP), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), benzyl alcohol, benzyl benzoate, and mixtures thereof, preferably N-methylpyrrolidone (NMP), dimethylacetamide (DMA), and dimethyl sulfoxide (DMSO). The composition may further comprise an aqueous medium (such as an aqueous buffer or saline) and / or a surfactant (such as a polyethylene glycol fatty acid ester, e.g., PEG-15 hydroxystearate). Advantageously, the long-acting injectable depot pharmaceutical composition comprises an aqueous suspension. Advantageously, the aqueous suspension is a nanosuspension. Advantageously, the long-acting injectable depot pharmaceutical composition does not contain a rate-controlling agent, such as a biodegradable polymer, or contains less than 20% by weight of the total composition of the rate-controlling agent, or less than 15%, less than 10%, less than 5%, less than 3%, less than 2%, or less than 1% by weight. Advantageously, the Compound 1 form is Form C.

[0158] In one embodiment, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. a) Compound 1 free form (conveniently, anhydrous Form A or Form C), and one or more pharmaceutically acceptable excipients 1. A long-acting injectable depot pharmaceutical composition comprising: the long-acting injectable depot pharmaceutical composition comprises 300 to 900 mg of Compound 1, the long-acting injectable depot pharmaceutical composition is administered in an injection volume of about 0.1 to about 2 mL, and the long-acting injectable depot pharmaceutical composition comprises an aqueous suspension, preferably an aqueous based nanosuspension; A long-acting injectable depot pharmaceutical composition is provided. Advantageously, Compound 1 Form is Form C.

[0159] In one embodiment, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. a) Compound 1 free form as anhydrous Form C, and b) one or more pharmaceutically acceptable excipients 1. A long-acting injectable depot pharmaceutical composition comprising: The long-acting injectable depot pharmaceutical composition comprises: i. containing 300 to 900 mg of compound 1; ii. Contains an aqueous suspension; iii. has an injection volume of about 0.1 to about 2 mL; and iv. sustainably releases Compound 1 at a rate that results in therapeutic plasma concentrations of Compound 1 for at least 6 months after subcutaneous administration; A long-acting injectable depot pharmaceutical composition is provided.

[0160] In one embodiment, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. a) Compound 1 free form as anhydrous Form C; and b) one or more pharmaceutically acceptable excipients 1. A long-acting injectable depot pharmaceutical composition comprising: The long-acting injectable depot pharmaceutical composition comprises: i. containing 300 to 900 mg of compound 1; ii. Contains an aqueous suspension; iii. has an injection volume of about 0.1 to about 2 mL; and iv. sustained release of Compound 1 at a rate that results in therapeutic plasma concentrations of Compound 1 for at least 6 months following intramuscular administration; A long-acting injectable depot pharmaceutical composition is provided.

[0161] Preferably, the long-acting injectable depot pharmaceutical composition comprising Compound 1 in an aqueous suspension may further comprise one or more of an isotonic agent (e.g., sodium chloride or mannitol), a suspending agent (e.g., polyvinylpyrrolidone or a polysorbate, e.g., polysorbate 80), and a surfactant (e.g., a poloxamer, e.g., poloxamer 188). Conveniently, the aqueous suspension is a microsuspension having a D10 of 0.3-2 μm, a D50 of 2-4 μm, and / or a D90 of 4-13 μm. Conveniently, the aqueous suspension is a nanosuspension having a D10 of 1-100 nm, a D50 of 1-150 nm, and / or a D90 of 1-200 nm.

[0162] In one embodiment, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. a) Compound 1, and b) one or more pharmaceutically acceptable excipients 1. A long-acting injectable depot pharmaceutical composition comprising: The long-acting injectable depot pharmaceutical composition comprises: i. containing 300 to 900 mg of compound 1; ii. containing a solution of Compound 1 in NMP; iii. has an injection volume of about 0.1 to about 2 mL; and iv. sustainably releases Compound 1 at a rate that results in therapeutic plasma concentrations of Compound 1 for at least 6 months after subcutaneous administration; A long-acting injectable depot pharmaceutical composition is provided.

[0163] Preferably, the long-acting injectable depot pharmaceutical composition containing Compound 1 in NMP solution may further contain one or more of an aqueous medium (such as an aqueous buffer or saline) and a surfactant (such as a polyethylene glycol fatty acid ester, e.g., PEG-15 hydroxystearate). Advantageously, the composition contains Compound 1 at a concentration of 100-400 mg / mL and NMP at 5-95% v / v.

[0164] Preparation of the Composition In a second aspect, the present invention provides a method of forming a pharmaceutical composition according to the first aspect.

[0165] In one embodiment, the pharmaceutical composition of the invention is an aqueous suspension depot. In one embodiment, the method for preparing the aqueous suspension depot comprises: a) incorporating a helicase-primase inhibitor or a pharmaceutically acceptable salt thereof into a water-based vehicle, wherein the water-based vehicle may contain a suspending agent (e.g., polysorbate 80 or poloxamer 188) and may also contain an isotonicity agent (e.g., sodium chloride); and b) mixing the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof (e.g., by stirring and / or vortexing and / or sonication) to provide a homogenous suspension suitable for subcutaneous or intramuscular administration. Includes:

[0166] In one embodiment, the pharmaceutical composition of the present invention is an aqueous based microsuspension depot. In one embodiment, the method for preparing the aqueous based microsuspension depot comprises: a) reducing the particle size of the active ingredient helicase-primase inhibitor or a pharmaceutically acceptable salt thereof, for example by wet or dry milling, to provide micro-sized particles (e.g., Dv90<10 μm); b) incorporating micro-sized particles of the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof into an aqueous vehicle, wherein the aqueous vehicle may contain a suspending agent (e.g., polysorbate 80 or poloxamer 188), an isotonicity agent (e.g., sodium chloride), and a viscosity modifier (e.g., hydroxypropylmethylcellulose); and c) mixing (e.g., by stirring and / or vortexing and / or sonication) micro-sized particles of the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof in an aqueous vehicle to provide a uniform suspension suitable for subcutaneous or intramuscular administration. Includes:

[0167] In one embodiment, the milling carried out in step a) is jet milling.

[0168] In one embodiment, the pharmaceutical composition of the present invention is an aqueous based nanosuspension depot. In one embodiment, the method for preparing the aqueous based nanosuspension depot comprises: a) reducing the particle size of the active ingredient helicase-primase inhibitor or a pharmaceutically acceptable salt thereof, for example by wet milling, to provide nano-sized particles (e.g., Dv90<500 nm); b) incorporating nano-sized particles of the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof into an aqueous vehicle, wherein the aqueous vehicle may contain a suspending agent (e.g., polysorbate 80 or poloxamer 188) and may also contain an isotonicity agent (e.g., mannitol), and may also contain an isotonicity agent (e.g., sodium carboxymethylcellulose); and c) mixing nano-sized particles of the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof in an aqueous vehicle (e.g., by stirring and / or vortexing and / or sonication) to provide a uniform nanosuspension suitable for subcutaneous or intramuscular administration. Includes:

[0169] In one embodiment, the milling carried out in step a) is wet milling at 700 rpm for at least 300 minutes to reduce the particle size to nano-sized particles, for example Dv90<500 nm.

[0170] In one embodiment, particle size reduction of the active ingredient helicase primase inhibitor or a pharmaceutically acceptable salt thereof to provide nano-sized particles (e.g., Dv<500 nm) for nanosuspension is performed in stages. For example, particles of the active ingredient helicase primase inhibitor or a pharmaceutically acceptable salt thereof are first reduced to micro-sized material, e.g., by a high-pressure homogenizer, until the size is approximately 1 μm. The method then includes milling this material, e.g., by wet milling using zirconium oxide milling beads, e.g., at 700 rpm for 500 minutes, to reduce the particle size to nano-sized particles (Dv<500 nm).

[0171] Therapeutic Uses and Applications The present invention provides a method for treating herpes virus (preferably HSV) infection in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a helicase primase inhibitor or a pharmaceutically acceptable salt thereof, wherein the helicase primase inhibitor or a pharmaceutically acceptable salt thereof is administered via an injection route. Advantageously, the helicase primase inhibitor or a pharmaceutically acceptable salt thereof is administered by subcutaneous or intramuscular injection.

[0172] The present invention provides a method for treating a herpes virus (conveniently HSV) infection in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, wherein the helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, is formulated as a pharmaceutical composition according to the first aspect of the present invention.

[0173] The present invention also provides a pharmaceutical composition according to the first aspect of the present invention, comprising a helicase-primase inhibitor or a pharmaceutically acceptable salt thereof for use as a pharmaceutical.

[0174] The present invention also provides a pharmaceutical composition according to the first aspect of the invention comprising a helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, for use in the treatment of a herpes virus (conveniently HSV) infection in a subject in need thereof.

[0175] The present invention also provides the use of a pharmaceutical composition according to the first aspect of the invention comprising a helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a herpes virus (conveniently HSV) infection in a subject in need thereof.

[0176] In certain embodiments, the present invention provides a method for inhibiting HSV replication in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a helicase-primase inhibitor or a pharmaceutically acceptable salt thereof, wherein the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof is formulated as a pharmaceutical composition according to the first aspect of the present invention.

[0177] In one embodiment, the present invention also provides a method for reducing the likelihood or severity of symptoms of HSV infection in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a helicase-primase inhibitor or a pharmaceutically acceptable salt thereof, wherein the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof is formulated as a pharmaceutical composition according to the first aspect of the present invention.

[0178] In one embodiment, the present invention provides a method for inhibiting the onset or progression of a disease or disorder caused by or associated with HSV infection in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, wherein the helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, is formulated as a pharmaceutical composition according to the first aspect of the present invention.

[0179] In one embodiment, the present invention provides a method for inhibiting the recurrence of HSV symptoms or outbreaks in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a helicase-primase inhibitor or a pharmaceutically acceptable salt thereof, wherein the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof is formulated as a pharmaceutical composition according to the first aspect of the present invention.

[0180] In one embodiment, the present invention provides a pharmaceutical composition according to the first aspect of the invention comprising a helicase-primase inhibitor or a pharmaceutically acceptable salt thereof for use in suppressing the recurrence of HSV symptoms or outbreaks in a subject in need thereof.

[0181] In one embodiment, the present invention provides use of a pharmaceutical composition according to the first aspect of the invention comprising a helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for suppressing the recurrence of HSV symptoms or outbreaks in a subject in need thereof.

[0182] In one embodiment, the present invention provides a method for treating or preventing a disease or disorder caused by or associated with HSV infection in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a helicase-primase inhibitor or a pharmaceutically acceptable salt thereof, wherein the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof is formulated as a pharmaceutical composition according to the first aspect of the present invention. In certain embodiments, the disease or disorder caused by or associated with HSV infection is selected from herpes labialis (e.g., cold sore or whitlow), genital herpes, HSV-associated keratitis, HSV-associated encephalitis, pneumonia, martial artist's herpes, primary HSV gingivostomatitis, Mollaret's meningitis, and Bell's palsy.

[0183] In certain embodiments, the disease or disorder caused by or related to HSV infection is selected from herpes labialis (cold sore or whitlow) or genital herpes.In one embodiment, the disease or disorder is recurrent herpes labialis or recurrent genital herpes.People who have a history of multiple recurrences of herpes labialis or recurrent genital herpes (for example, HSV recurrences six or more times per year) can be considered to suffer from recurrent HSV.

[0184] In one embodiment, the herpes virus being treated is HSV 2. In a further embodiment, the herpes virus being treated is HSV 2 and the subject in need of treatment has HSV 2 recurrent genital herpes.

[0185] In one embodiment, the herpes virus being treated is HSV 1. In a further embodiment, both herpes viruses HSV 1 and HSV 2 are being treated.

[0186] In one embodiment, the herpes virus being treated is resistant to a nucleoside antiviral therapy, hi one embodiment, the nucleoside antiviral therapy is selected from the group consisting of acyclovir, penciclovir, famciclovir, ganciclovir, and valacyclovir.

[0187] In one embodiment, the herpes virus infection being treated is resistant to nucleoside antiviral therapy, e.g., an acyclovir-resistant mucocutaneous HSV infection. In a further embodiment, the HSV infection being treated is a mucocutaneous HSV infection that is resistant to treatment with a nucleoside analog antiviral therapy, such as acyclovir, penciclovir, famciclovir, ganciclovir, or valacyclovir.

[0188] In certain embodiments, a subject in need of the methods disclosed herein is immunocompromised. The subject may be immunocompromised due to conditions including HIV infection, cancer, hematopoietic cell or solid organ transplant, chronic glucocorticoid use, or genetic immunodeficiency.

[0189] In certain embodiments, the subject in need of the methods disclosed herein is a newborn or infant.

[0190] In certain embodiments, the subject is a herpes positive patient.

[0191] In certain embodiments, a subject in need of the methods disclosed herein has an acyclovir-resistant mucocutaneous HSV infection, which can be diagnosed based on clinical failure, e.g., failure to improve after at least 7 days of oral or iv administration of approved doses of acyclovir.

[0192] In certain embodiments, a subject in need of the methods disclosed herein has a primary genital HSV-associated herpes infection. In one embodiment, a subject in need of the methods disclosed herein has a severe or progressive genital HSV-associated herpes infection.

[0193] In certain embodiments, the pharmaceutical composition according to the first aspect of the present invention can reduce the recurrence of HSV infections (i.e., provide suppressive therapy) that cause diseases or disorders such as herpes labialis or genital herpes. In one embodiment, the reduction in the number of recurrences of lesions over a one-year period can be 20% or more, 30% or more, 40% or more, 50% or more, 75% or more, 90% or more, or 95% or more. Advantageously, the reduction in lesion incidence over a one-year period can be 90% or more.

[0194] In certain embodiments, the pharmaceutical composition according to the first aspect of the invention is capable of reducing the duration of recurrent episodes of HSV infection by, for example, 1 day or more, such as by at least 2 days, 3 days, 4 days, 5 days, 14 days, 21 days or 28 days.

[0195] In certain embodiments, the pharmaceutical composition according to the first aspect of the present invention may reduce the time to healing (e.g., the time to complete resolution of lesions) and the duration of symptoms resulting from HSV infection in diseases or disorders such as herpes labialis or genital herpes. Time to lesion healing may be defined as complete epithelialization of mucocutaneous HSV lesions and the absence of new lesions within the treatment period, as assessed, for example, by a physician.

[0196] In one embodiment, the pharmaceutical composition according to the first aspect of the present invention is capable of reducing pain or pain intensity (e.g., at the site of a lesion) caused as a result of HSV infection in diseases or disorders such as herpes labialis or genital herpes.

[0197] In certain embodiments, the pharmaceutical composition according to the first aspect of the present invention can reduce or slow the rate of viral shedding in individuals with frequently recurring HSV, such as genital HSV2. For example, the rate of genital HSV mucocutaneous shedding in a subject can be measured by taking swabs of the skin and mucous membranes and analyzing the samples for HSV DNA, for example, with a real-time quantitative fluorescent polymerase chain reaction (PCR) assay, for HSV detection. The frequency of HSV2 detection (viral shedding rate) can be defined as the number of days on which a genital swab is positive for HSV divided by the total number of days on which a genital swab is obtained. The reduction in HSV shedding rate among subjects administered the compositions of the present invention can be compared to the shedding rate among subjects receiving a placebo or other treatment. The amount of HSV in positive swabs, as well as the frequency of genital lesions and shedding episodes, can also be monitored.

[0198] In certain embodiments, the present invention provides a method for reducing (or substantially inhibiting or eliminating) disruptive HSV shedding in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, wherein the helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, is formulated as a pharmaceutical composition according to the first aspect of the present invention.

[0199] In one embodiment, the present invention provides a pharmaceutical composition according to the first aspect of the invention comprising a helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, for use in reducing (or substantially inhibiting or eliminating) disruptive HSV shedding in a subject in need thereof.

[0200] In one embodiment, the present invention provides use of a pharmaceutical composition according to the first aspect of the invention comprising a helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for reducing (or substantially suppressing or eliminating) disruptive HSV shedding in a subject in need thereof.

[0201] In certain embodiments, the present invention provides a method for preventing infection with an infection caused by HSV, the method comprising administering to a subject a therapeutically effective amount of a helicase-primase inhibitor or a pharmaceutically acceptable salt thereof, wherein the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof is formulated as a pharmaceutical composition according to the first aspect of the present invention.

[0202] In certain embodiments, the present invention provides a method for reducing side effects observed when a helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, is administered by oral administration to a subject having an infection caused by HSV, the method comprising administering to the subject a therapeutically effective amount of the helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, wherein the helicase-primase inhibitor, or a pharmaceutically acceptable salt thereof, is formulated as a pharmaceutical composition according to the first aspect of the present invention.

[0203] The pharmaceutical compositions of the present invention are injectable depot compositions and are therefore administered intramuscularly or subcutaneously. In a preferred embodiment, the pharmaceutical compositions of the present invention are administered subcutaneously. In a preferred embodiment, the pharmaceutical compositions of the present invention are administered intramuscularly.

[0204] When used according to this embodiment, the appropriate dosage will vary depending, for example, on the nature and severity of the infection being treated and is within the purview of the treating physician. Typically, the recommended dosage may range from about 0.1 to about 1,000 μg / kg body weight. In some cases, the compound may be administered at a dosage of less than 400 μg / kg body weight. In other cases, the dosage may be less than 200 μg / kg body weight. In still other cases, the dosage may range from about 0.1 to about 100 μg / kg body weight. In one embodiment, the therapeutically effective amount of the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof is from about 5 mg to about 900 mg, e.g., from about 100 mg to about 600 mg.

[0205] Advantageously, applicants have discovered that the extended release provided by the compositions of the present invention, and the resulting stable plasma levels that remain at therapeutically effective plasma concentrations for an extended period of time after administration, allows for the potential use of lower doses than expected.

[0206] Doses may conveniently be administered no more than twice a month, no more than once a month, no more than once every two months, no more than once every three months, no more than once every four months, no more than once every five months, or no more than once every six months. Advantageously, doses may be administered once a month, once every two months, or once every three months.

[0207] In a preferred embodiment, the unit dosage form of the pharmaceutical composition is a subcutaneous injection containing about 5 mg to about 900 mg (e.g., about 100 mg to about 900 mg) of the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof. In a most preferred embodiment, the unit dosage form of the pharmaceutical composition is a subcutaneous injection containing about 100 mg to about 600 mg of the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof.

[0208] In a preferred embodiment, the unit dosage form of the pharmaceutical composition contains about 100 mg to about 1200 mg (e.g., about 100 mg to about 900 mg) of Compound 1 or a pharmaceutically acceptable salt thereof. In a most preferred embodiment, the unit dosage form of the pharmaceutical composition is a subcutaneous injection containing about 100 mg to about 900 mg of Compound 1 or a pharmaceutically acceptable salt thereof. In a more preferred embodiment, the unit dosage form of the pharmaceutical composition is a subcutaneous injection containing about 200 mg to about 900 mg of Compound 1 or a pharmaceutically acceptable salt thereof, for example, about 200 mg to about 600 mg, about 300 mg to about 900 mg, or about 300 mg to about 600 mg.

[0209] In one embodiment, the present invention provides a method for treating a herpes virus (conveniently HSV) infection in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a helicase-primase inhibitor or a pharmaceutically acceptable salt thereof, wherein the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof is formulated as a pharmaceutical composition according to the first aspect of the present invention (e.g., certain long-acting injectable depot pharmaceutical compositions including an organic solvent solution or a microsuspension or an aqueous nanosuspension), and wherein no loading dose, such as an oral loading dose, is required prior to administration of the long-acting injectable depot pharmaceutical composition according to the first aspect of the present invention.

[0210] Use in combination The pharmaceutical compositions of the present invention can be administered alone as a monotherapy or can be administered in combination with one or more additional substances and / or treatments, which can be achieved by simultaneous, sequential or separate administration of the individual therapeutic components.

[0211] Also contemplated herein are methods that include administering a second active agent. For example, in addition to being infected with HSV, the subject or patient may also have an HSV infection-related comorbidity, i.e., a disease or other adverse health condition that is associated with, aggravated by, or caused by HSV infection. Also contemplated herein are the disclosed pharmaceutical compositions in combination with at least one other agent previously shown to treat these HSV infection-related conditions. Such combined treatment can be achieved independently (by simultaneous, sequential, or separate administration of the individual therapeutic components) and / or via the pharmaceutical composition of the present invention that includes a second active agent.

[0212] Accordingly, provided herein is a method for treating or preventing an HSV infection in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a helicase-primase inhibitor or a pharmaceutically acceptable salt thereof, wherein the helicase-primase inhibitor or a pharmaceutically acceptable salt thereof is formulated as a pharmaceutical composition according to the first aspect of the present invention, and co-administering to the subject a therapeutically effective amount of an additional therapeutic agent.

[0213] In one embodiment, the additional therapeutic agent is one of the following agents: i. Nucleoside polymerase inhibitors, such as acyclovir, valacyclovir, famciclovir, penciclovir, ganciclovir; ii. Pyrophosphate polymerase inhibitors, such as foscarnet; iii. saturated fatty alcohols, such as docosanol; iv. Drugs such as idoxuridine, trifluridine and vidarabine; v. Corticosteroids; and vi. Other helicase-primase inhibitors, such as amenamevir One or more of the following is selected.

[0214] In some cases, the disclosed pharmaceutical compositions according to the first aspect of the present invention can be administered as part of a combination therapy with one or more antiviral agents, including nucleoside analogues such as acyclovir, foscarnet, ganciclovir or penciciovir or their respective prodrugs valacyclovir or famciclovir.

[0215] In some embodiments, the first amount and the second amount together constitute a pharmaceutically effective amount.The first amount, the second amount, or both may be the same as, greater than, or less than the effective amount of each compound administered as a monotherapy.The therapeutically effective amounts of the disclosed compound and antiviral agent may be co-administered to a subject, i.e., may be administered to a subject simultaneously or separately, in any predetermined order, by the same or different administration routes.In some cases, it may be advantageous to start administering Compound 1 first, for example, one or more days or several weeks before the start of administration of the antiviral agent.In addition, additional drugs may be administered in combination with the above-mentioned combination therapy.

[0216] kit In one embodiment, the pharmaceutical compositions and methods described herein provide kits for treating disorders such as those described herein. These kits include a pharmaceutical composition described herein in a container and, optionally, instructions for using the kit according to the various methods and approaches described herein. Such kits may also include information, such as scientific literature, package inserts, clinical trial results, and / or summaries thereof, that demonstrate or establish the activity and / or benefits of the composition and / or describe dosage, administration, side effects, drug interactions, or other information useful to healthcare providers. Such information may be based on the results of various studies, such as studies using laboratory animals, including in vivo models, and studies based on human clinical trials. The kits described herein may be provided, sold, and / or promoted to healthcare providers, including physicians, nurses, pharmacists, prescribers, and the like. Kits may also, in some embodiments, be sold directly to consumers.

[0217] The pharmaceutical compositions of the present invention can be utilized for diagnostic purposes and as research tools.

[0218] In addition to being useful for human treatment, the pharmaceutical compositions of the present invention may be useful for the veterinary treatment of companion animals, exotic animals and farm animals, including mammals, rodents, etc. Advantageously, such animals include horses, dogs and cats.

[0219] The invention is illustrated by the following non-limiting examples. [Example]

[0220] The following abbreviations are used herein: ACN: acetonitrile DCM: dichloromethane DMF: dimethylformamide DMSO: dimethyl sulfoxide DSC: Differential scanning calorimetry DVS: Dynamic Vapor Sorption EDCI: 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide eq: equivalent EtOAc: ethyl acetate hr: hour HPLC: High-performance liquid chromatography K2CO3: Potassium carbonate K3PO4: Potassium phosphate KF: Karl Fischer titration MeOH: Methanol min: minutes NaCMC: sodium carboxymethylcellulose Na2SO4: Sodium sulfate NMP: N-methylpyrrolidone NMT: Below PLM: Polarized Light Microscope PVP K30: Polyvinylpyrrolidone K30 grade (Povidone K30) RH: Relative humidity RT: Room temperature (approx. 22℃) TBAB: Tetrabutylammonium bromide TGA: Thermogravimetric analysis THF: tetrahydrofuran TLC: thin layer chromatography Vol: Volume XRPD: X-ray powder diffraction

[0221] material and method Particle size distribution by light diffraction The particle size distributions of the suspensions prepared in Examples 1 and 10 were measured by laser diffraction using a NANOTRAC FLEX particle size analyzer (Microtrac Instruments). Water was used as the medium. Samples were added to maintain a laser obscuration level between 0.1 and 10%. A measurement time of 20 seconds was used, and a background measurement was performed using water for 10 seconds. Dv 10 , Dv 50 and Dv 90 The diameter of the was measured and is reported below.

[0222] The particle size distribution of the suspensions prepared in Examples 14 and 17 was measured using a Mastersizer 3000 particle size analyzer (Malvern, UK). The suspensions were freeze-dried, and the freeze-dried powder was used to prepare a 0.1% suspension in water for testing. Water was used as the dispersion medium. The suspension was added until the final obscuration range was 3-10%. Each sample was measured three times in total. Dv 10 , Dv 50 and Dv 90 The diameter of the was measured and is reported below.

[0223] scanning electron microscope Scanning electron microscope (SEM) images of the suspensions prepared in Examples 14 and 17 below were taken using an Oxford X-MAS scanning electron microscope (Zeiss-Sigma-HD, Germany). Prior to measurement, the suspensions were diluted with distilled water (4x) and sonicated for approximately 5 minutes. The samples were then immersed in conductive adhesive and air-dried at room temperature. Loose powder was removed using a rubber suction bulb. The samples were then sprayed with gold and inspected.

[0224] LC-MS analysis of plasma samples The LC-MS method and conditions used to analyze plitelivir in plasma samples obtained from the PK studies described in Examples 1-5 are shown in Table 1.

[0225] [Table 1]

[0226] The LC-MS method and conditions used for the analysis of plitelivir in dog plasma samples obtained from the PK study described in Example 14 are shown in Table 2.

[0227] [Table 2]

[0228] The LC-MS method and conditions used to analyze helicase-primase inhibitor Compound 1 in plasma samples obtained from the subcutaneous injection PK studies (Example 7 and Examples 10-12) are described in Tables 3 and 4.

[0229] [Table 3]

[0230] [Table 4]

[0231] The LC-MS method and conditions used to analyze Compound 1 in plasma samples obtained from the intravenous injection PK study (Example 13) are listed in Tables 5-7.

[0232] [Table 5]

[0233] [Table 6]

[0234] [Table 7]

[0235] Table 8 shows the LC-MS method and conditions used to analyze helicase-primase inhibitor Compound 1 in rat plasma samples obtained from the subcutaneous injection PK study described in Example 15.

[0236] [Table 8]

[0237] Table 9 shows the LC-MS method and conditions used to analyze helicase-primase inhibitor Compound 1 in dog plasma samples obtained from the dog PK study described in Example 16.

[0238] [Table 9]

[0239] The LC-MS method and conditions used to analyze Compound 1 in rat plasma samples obtained from the PK study described in Example 17 are shown in Table 10.

[0240] [Table 10]

[0241] X-ray powder diffraction (XRPD) analysis Equipment: Rigaku Smartlab SE X-ray wavelength: Cu, Kα, Kα1(Å):1.540598, Kα2(Å):1.544426 Kα2:Kα1 intensity ratio: 0.50 X-ray tube settings: 40kV, 15mA, Scan Mode 1D Scan range (2θ): 3° to 40°, step size (2θ): 0.02° (scan speed (2θ) 10° / min)

[0242] Priterivir free base (dihydrate) Plitelivir (100 mg) was dissolved in 88:12 THF / water (1 mL) at 60° C. The solution was cooled and water (4 mL) was added. The resulting suspension was filtered and the solid was collected and dried.

[0243] The crystalline plitelivir free base obtained according to this procedure was analyzed as described below and used in Examples 1-6 described herein.

[0244] Pritelivir free base was analyzed by TGA and XRPD. By TGA, the material lost 7.9% of its weight as water between 30°C and 90°C, corresponding to a dihydrate. The XRPD diffractogram is shown in Figure 1, and the peak positions are listed in the table below. Characteristic peaks include those at 11.0° 2θ, 11.3° 2θ, and 21.1° 2θ. [Table 11]

[0245] Priterivir free base (sesquihydrate) 2-(4-(pyridin-2-yl)phenyl)acetic acid (38.40 g, 80%, 180.074 mmol, 1.00 equivalents) was added to DMF (385 mL). HOBT (17.38 g, 128.624 mmol, 0.71 equivalents) was added at 20° C., followed by stirring for 10 minutes. 2-Chloro-4-methylthiazole-5-sulfonamide (26.66 g, 128.624 mmol, 0.71 equivalents) and EDCI (27.12 g, 141.486 mmol, 0.79 equivalents) were added. The mixture was stirred at 20° C. under N for 36 hours. The mixture was added to water (780 mL) and stirred for 30 minutes. The solid was collected by filtration. The filter cake was washed twice with water (100 mL x 2) and then dried under reduced pressure at 50°C for 12 hours to give plitelivir as a white solid (yield 78.4%).

[0246] The crystalline plitelivir free base obtained according to this procedure was analyzed as described below and used in Example 14.

[0247] Plitelivir free base was analyzed by TGA and XRPD. By TGA, the material lost 5.6% weight between 35 and 101°C. Characterization data suggests that it is a sesquihydrate of plitelivir (1.5 water molecules per plitelivir molecule). The XRPD diffractogram is shown in Figure 16, and the peak positions are listed in the table below. Characteristic peaks include those at 12.1, 14.6, and 18.3° 2θ.

[0248] [Table 12]

[0249] Compound 1 Free Base (Anhydrous Form A) Crude compound 1 (36.5 g), prepared according to Example 8, was stirred in DMSO (300 mL, 8.2 rel. vol.) at 15-27°C for 20-30 minutes to form a solution. Water (1100 mL, 30.1 rel. vol.) was added slowly over 30 minutes, and the mixture was stirred at 15-27°C for 1-2 hours. Additional water (1820 mL, 49.9 rel. vol.) was added slowly over 60 minutes, and the mixture was stirred at 15-27°C for 12 hours. The mixture was then filtered, washed with water (365 mL x 3, 10 vol. x 3), and the cake was dried at 45-55°C to give compound 1 (26.4 g, 72% yield) as an off-white solid.

[0250] Crystalline Compound 1 free base Form A obtained according to this procedure was analyzed as described below and used in Examples 7 and 10-12.

[0251] The resulting crystalline form had the XRPD diffractogram shown in Figure 17 and was designated Form A. The peak positions present in the XRPD diffractogram obtained for Form A are shown in the table below.

[0252] [Table 13]

[0253] Further solid-state characterization of Form A using DSC and TGA revealed that Form A is an anhydrous crystalline form of Compound 1.

[0254] Compound 1 Free Base (Anhydrous Form C) Crude Compound 1 (4.0 kg) prepared according to Example 8 was stirred in DMSO (12 L, 3 rel. vol.) at 75-85°C in a first reactor to form a solution. The solution was then filtered through a 0.2 μm filter into a pre-warmed (75-85°C) second reactor, after which the first reactor was rinsed with DMSO (2 L, 0.5 rel. vol.) and the rinse was transferred to the second reactor. The contents of the second reactor were stirred at 75-85°C for 30-60 minutes and then cooled at a rate of 8-12°C / hour to obtain a batch temperature of 50-55°C. The mixture was stirred at 50-55°C for 1-2 hours. Ethanol (4 L, 1 rel. vol.) was then slowly added over 60-90 minutes while maintaining the batch temperature at 50-55°C. The internal batch temperature was raised to 57-62°C, and ethanol (52 L, 13 rel. vol.) was slowly added over 7.0-7.5 hours. The contents were stirred at 57-62°C for 1-2 hours, then cooled at a cooling rate of 8-12°C / hour to obtain a final batch temperature of 18-22°C. The mixture was stirred at 18-2°C for 8-12 hours, then filtered, washed with ethanol (8 L, 2 vol.), and the cake was dried at 45-55°C for 20-24 hours to give 3.6 kg (90% yield) of compound 1 as an off-white to pale yellow solid.

[0255] Crystalline Compound 1 free base obtained according to this procedure was analyzed as described below and used in Example 17.

[0256] The resulting crystalline form had the XRPD diffractogram shown in Figure 18, which was designated as Form C. The peak positions present in the XRPD diffractogram obtained for Form C are shown in the table below:

[0257] [Table 14]

[0258] Further solid-state characterization of Form C was performed using DSC, TGA, and DVS. Form C was found to be an anhydrous crystalline form of Compound 1.

[0259] Antiviral assay In vitro EC of helicase-primase inhibitors against HSV-1 and HSV-2 50 The following assay can be used to measure

[0260] Cytopathic effect (CPE) inhibition assay Cell Culture—Vero cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal bovine serum and 100 units / mL penicillin and streptomycin. The cells were passaged 2–3 times per week to maintain subconfluent density.

[0261] HSV-1 antiviral assay - Vero cells, 2.5 x 10 cells per well in a 96-well plate 3 Cells were seeded at a density of 1 / 3 and allowed to attach overnight. After attachment, the medium was replaced with 50 μL of infection medium (DMEM supplemented with 2% fetal bovine serum and 100 units / mL penicillin and streptomycin). Compounds were then added to the cultures in an 8-point, 3-fold serial dilution using a Tecan D300e digital dispenser. DMSO concentrations were normalized to 0.5% for all treatments. After compound addition, 50 μL of infection medium containing 80 TCID50 HSV-1 was added to the cells and incubated at 37°C for 4 days. After incubation, the plates were equilibrated to room temperature, the medium was removed, and 60 μL of a 1:1 dilution of Cell titer glow in phosphate-buffered saline was added. After a 5-minute incubation, cell viability was quantified by measuring brightness using a Tecan Infinite M1000 Pro plate reader.

[0262] HSV-2 antiviral assay - Vero cells, 1.0 x 10 cells per well in a 96-well plate 4 Cells were seeded at a density of 100 μL per well and allowed to attach overnight. After attachment, the medium was replaced with 50 μL of infection medium (DMEM supplemented with 2% fetal bovine serum and 100 units / mL penicillin and streptomycin). Compounds were then added to the cultures in an 8-point, 3-fold serial dilution using a Tecan D300e digital dispenser. DMSO concentrations were normalized to 0.5% for all treatments. After compound addition, 50 μL of infection medium containing 160 TCID50 HSV-2 G strain was added to the cells and incubated at 37°C for 5 days. After incubation, 10 μL / well of WST-8 chromogenic reagent was added, and the plates were incubated at 37°C for 3 hours. After incubation, cell viability was quantified by measuring absorbance at 460 nm and 620 nm using a Tecan Infinite M1000 Pro plate reader.

[0263] Using this CPE assay, the mean EC 50 The values ​​were 0.047 μM and 0.055 μM, respectively.

[0264] Example 1: Plitelivir PK study in rats Preparation of formulations Aqueous suspensions of crystalline plitelivir free base (dihydrate) in 0.9% NaCl + 0.5% polysorbate 80 vehicle (concentrations of 10.0 mg / mL and 50.0 mg / mL) were prepared as follows: 1) Vehicle (0.9% w / w NaCl and 0.5% w / w polysorbate 80) was prepared by dissolving NaCl and polysorbate 80 in water. 2) An appropriate amount of crystalline plitelivir free base (dihydrate) was weighed and added to the vehicle. 3) The mixture was vortexed for 1 minute, stirred for 10 minutes, and then sonicated for 15 minutes. 4) The final formulation was a uniform, cloudy white suspension.

[0265] Formulation characterization The particle size distribution (PSD) of the suspension is Dv 10 =1.45μm, Dv 50 = 4.23 μm, and Dv 90 = 10.35 μm.

[0266] Animal administration For the pharmacokinetic study, male Sprague Dawley (SD) rats (6-8 weeks old) weighing approximately 300 g were used. Animals had free access to food and water. The pritelivir suspension was administered subcutaneously to each group of animals (n = 3) at doses of 33.33 mg / kg and 166.67 mg / kg in a volume of 3.33 mL / kg.

[0267] Approximately 200 μL of whole blood was collected from the jugular vein into K2EDTA tubes at each time point. The blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. The plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0268] result The mean plasma concentration profiles (n = 3) after subcutaneous (SC) administration of pritelivir suspension at two doses (33.33 mg / kg and 166.67 mg / kg) are shown in Figure 2. Prolonged exposure out to at least 720 hours (study duration) was achieved at both dose levels.

[0269] Example 2: Plitelivir PK study at low doses in rats Preparation of formulations An aqueous suspension of crystalline plitelivir free base (dihydrate) in 0.9% NaCl + 0.5% polysorbate 80 vehicle (concentration 3.0 mg / mL) was prepared as follows: 1) Vehicle (0.9% w / w NaCl and 0.5% w / w polysorbate 80) was prepared by dissolving NaCl and polysorbate 80 in water. 2) An appropriate amount of crystalline plitelivir free base (dihydrate) was weighed and added to the vehicle. 3) The mixture was stirred for 15 minutes, then vortexed for 1 minute and sonicated for 10 minutes. 4) The final formulation was a uniform, cloudy white suspension.

[0270] Animal administration For the rat PK study, male SD rats (6-8 weeks old) weighing 230-250 g were used. Animals had free access to food and water. The prepared suspension was administered subcutaneously to each group of animals (n = 3) at a dose of 10 mg / kg in a volume of 3.33 mL / kg.

[0271] Approximately 200 μL of whole blood was collected from the jugular vein into K2EDTA tubes at each time point. The blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. The plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0272] result The mean plasma concentration profiles (n = 3) after subcutaneous (SC) administration of pritelivir suspension are shown in Figure 3. At the low dose level of 10 mg / kg, prolonged exposure was achieved for at least 144 hours (study period).

[0273] Example 3: Plitelivir PK study in monkeys Preparation of formulations An aqueous suspension of crystalline plitelivir free base (dihydrate) in 0.9% NaCl + 0.5% polysorbate 80 vehicle (concentration 3.0 mg / mL) was prepared as follows: 1) Vehicle (0.9% w / w NaCl and 0.5% w / w polysorbate 80) was prepared by dissolving NaCl and polysorbate 80 in water. 2) An appropriate amount of crystalline plitelivir free base (dihydrate) was weighed and added to the vehicle. 3) The mixture was stirred for 15 minutes, then vortexed for 1 minute and sonicated for 10 minutes. 4) The final formulation was a uniform, cloudy white suspension.

[0274] Animal administration For the monkey PK study, male non-naive cyno monkeys weighing approximately 5 kg were used. Animals had free access to food and water. The prepared suspension was administered subcutaneously to each animal group (n = 3) at a dose of 10 mg / kg in a volume of 3.33 mL / kg.

[0275] Approximately 0.5 mL of whole blood was collected from the cephalic vein into K2EDTA tubes at each time point. The blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. The plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0276] result The mean plasma concentration profiles (n=3) after subcutaneous (SC) administration of pritelivir suspension are shown in Figure 4. Prolonged exposure was achieved at the 10 mg / kg dose level for at least 456 hours (study duration).

[0277] Example 4: Plitelivir PK study in dogs Preparation of formulations An aqueous suspension of crystalline plitelivir free base (dihydrate) in 0.9% NaCl + 0.5% poloxamer 188 vehicle (concentration 5.0 mg / mL) was prepared as follows: 1) Vehicle (0.9% w / w NaCl and 0.5% w / w Poloxamer 188) was prepared by dissolving NaCl and Poloxamer 188 in water. 2) An appropriate amount of crystalline plitelivir free base (dihydrate) was weighed and added to the vehicle. 3) The mixture was stirred for 15 minutes, then vortexed for 1 minute and sonicated for 10 minutes. 4) The final formulation was a uniform, cloudy white suspension.

[0278] Animal administration For the canine PK study, male non-naive beagles were used. The animals had free access to food and water. The prepared suspension was administered subcutaneously to each group of animals (n = 3) at a dose of 10 mg / kg in a volume of 2 mL / kg.

[0279] Approximately 0.5 mL of whole blood was collected into K2EDTA tubes at each time point. Blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. Plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0280] result The mean plasma concentration profiles (n=3) following subcutaneous (SC) administration of pritelivir suspension are shown in Figure 5. After an initial drug burst, prolonged exposure was achieved at the administered dose level.

[0281] Summary from PK studies In three species, a long-acting injectable depot formulation containing pritelivir was found to provide sustained and stable plasma concentrations of the drug for a significantly extended period after a single subcutaneous administration. Furthermore, prolonged release was achieved at different dose levels.

[0282] Additionally, no adverse reactions were observed at the injection site in the test animals.

[0283] Example 5: Comparative plitelivir PK study in rats after IV / SC administration Preparation of formulations A solution of crystalline plitelivir free base (dihydrate) in 5% NMP, 5% Solutol HS15, and 90% saline (concentration 0.5 mg / mL) was prepared as follows: 1) A 10 mg / mL stock solution of plitelivir (dihydrate) in NMP was prepared. 2) Equal volumes of Priteribiru stock solution and Solutol HS15 were combined and mixed. 3) The above solution was diluted with saline to a target pritelivir concentration of 0.5 mg / mL. The formulation was a clear solution.

[0284] Animal administration For the pharmacokinetic study, male Sprague Dawley (SD) rats (6-8 weeks old) weighing approximately 300 g were used. Animals had free access to food and water. The above pritelivir solution was administered intravenously to animals (n = 3) at a dose volume of 2 mL / kg to achieve a dose of 1 mg / kg. For subcutaneous injection, a dose volume of 10 mL / kg was used to achieve a dose of 5 mg / kg.

[0285] At each time point, approximately 200 μL of whole blood was collected from the jugular vein into K2EDTA tubes. Blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. Plasma samples were stored at approximately -70°C until analyzed by LC-MS according to the method described above.

[0286] result The mean plasma concentration profiles (n=3) after intravenous (IV) administration of 1 mg / kg pritelivir solution and subcutaneous (SC) administration of 5 mg / kg pritelivir solution can be seen in Figure 6. The calculated F for the SC route is approximately 117% compared to the IV route (Table below).

[0287] [Table 15]

[0288] This rat PK study demonstrates that the SC route of administration results in good exposure of pritelivir, with SC bioavailability surprisingly comparable to IV bioavailability.

[0289] Example 6: Stability and solubility studies of plitelivir suspension (dihydrate) Preparation of formulations An aqueous suspension of crystalline plitelivir free base (dihydrate) in 0.9% NaCl + 0.5% polysorbate 80 vehicle was prepared as follows: 1) Vehicle (0.9% w / w NaCl and 0.5% w / w polysorbate 80) was prepared by dissolving NaCl and polysorbate 80 in water. 2) An excess amount of plitelivir (dihydrate) was weighed and added to the vehicle to form a suspension of approximately 3 mg / mL. 3) The suspension was sonicated for 15 minutes and then stirred at 500 rpm at room temperature. 4) At 24 hours, the suspension was filtered, the filter cake was dried and analyzed by XRPD, and the filtrate was analyzed by HPLC to determine solubility.

[0290] HPLC analysis The HPLC method and conditions used for the quantification of plitelivir in the filtrate are listed in Table 11.

[0291] [Table 16]

[0292] result [Table 17]

[0293] XRPD analysis of the residual solid after the solubility experiment confirmed it to be the dihydrate (i.e., no polymorphic conversion occurred during the experiment).

[0294] Example 7: PK study using helicase-primase inhibitor compound 1 An additional helicase-primase inhibitor (HPI), Compound 1 (described in Example 8), was also subjected to animal PK studies after being formulated as a long-acting injectable depot composition according to the present invention.

[0295] Compound 1 was isolated as a crystalline solid and had an in vitro mean EC of 0.015 μM and 0.013 μM against HSV-1 and HSV-2, respectively, in the CPE assay described above. 50 had value.

[0296] PK study in rats Preparation of formulations As described in Example 2, an aqueous suspension of crystalline Compound 1 (Form A) in 0.9% NaCl+0.5% polysorbate 80 vehicle was prepared at a concentration of 3.0 mg / mL.

[0297] According to the animal administration method of Example 2, this suspension was administered to male SD rats at 10 mg / kg.

[0298] result The mean plasma concentration profiles (n=3) following subcutaneous (SC) administration of 10 mg / kg Compound 1 suspension are shown in Figure 7. When Compound 1 was formulated as an aqueous suspension, prolonged exposure was achieved at 10 mg / kg for at least 144 hours (study time).

[0299] PK study in monkeys Preparation of formulations As described in Example 3, an aqueous suspension of crystalline Compound 1 (Form A) in 0.9% NaCl+0.5% polysorbate 80 vehicle was prepared at a concentration of 3.0 mg / mL.

[0300] Following the animal administration method of Example 3, this suspension was administered to male cynomolgus monkeys at 10 mg / kg.

[0301] result The mean plasma concentration profiles (n=3) following subcutaneous (SC) administration of 10 mg / kg Compound 1 suspension are shown in Figure 8. When Compound 1 was formulated as an aqueous suspension, prolonged exposure was achieved at 10 mg / kg for at least 1392 hours (test time).

[0302] PK study in dogs Preparation of formulations As described in Example 4, an aqueous suspension of crystalline Compound 1 (Form A) in 0.9% NaCl + 0.5% Poloxamer 188 vehicle was prepared at a concentration of 5.0 mg / mL.

[0303] Following the animal administration method of Example 4, this suspension was administered at 10 mg / kg to male non-naive beagle dogs.

[0304] result The mean plasma concentration profiles (n=3) following subcutaneous (SC) administration of 10 mg / kg Compound 1 suspension are shown in Figure 9. When Compound 1 was formulated as an aqueous suspension, prolonged exposure was achieved at 10 mg / kg for at least 1392 hours (test duration).

[0305] Summary of PK studies using Compound 1 in Example 7 In three species, long-acting injectable depot formulations containing Compound 1 were found to provide sustained and stable plasma concentrations of the drug for a significantly extended period after a single subcutaneous administration. Furthermore, extended release was achieved at different dose levels. Furthermore, no adverse reactions were observed at the injection site in the test animals.

[0306] Example 8: Synthesis of 2-(3-(2',5'-difluoro-[1,1'-biphenyl]-4-yl)-2-oxotetrahydropyrimidin-1(2H)-yl)-4-methylthiazole-5-sulfonamide (Compound 1) [ka]

[0307] Synthesis of 1-(4-methylthiazol-2-yl)tetrahydropyrimidin-2(1H)-one (1-2) A mixture of compound 1-1 (6 g, 52.632 mmol) and 1-chloro-3-isocyanatopropane (6.26 g, 52.632 mmol) in THF (60 mL) was heated at 70 °C for 6 hours. TBAB (1.7 g, 5.263 mmol) and K2CO3 (18.15 g, 131.58 mmol) were added dropwise to the resulting solution while maintaining the same temperature, and the mixture was stirred at 70 °C for 16 hours. After the reaction was completed (monitored by TLC), the reaction mixture was diluted with water and extracted with EtOAc. The combined organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude compound was purified by CombiFlash chromatography (eluted with 60 to 70% EtOAc in heptane) to give the title compound 1-2 (5.1 g, 49.22%) as an off-white solid. TLC: 70% EtOAc / heptane (R f :0.5). Chemical formula C8H 11 MS calculated for N3OS: 197.06; Found: 198.17 [M+1] + . 1 H NMR (400 MHz, DMSO-d6) δ 7.30 (s, 1H), 6.60 (s, 1H), 3.99 (t, J = 5.4 Hz, 2H), 3.20 - 3.19 (m, 2H), 2.28 (s, 3H), 1.99 - 1.89 (m, 2H).

[0308] Synthesis of 1-(2',5'-difluoro-[1,1'-biphenyl]-4-yl)-3-(4-methylthiazol-2-yl)tetrahydropyrimidin-2(1H)-one (1-3) To a stirred solution of compound 1-2 (5 g, 25.380 mmol) in 1,4-dioxane (100 mL) was added intermediate 1A (8.16 g, 30.456 mmol), K2CO3 (8.75 g, 63.45 mmol), and then CuI (0.96 g, 5.076 mmol), and the resulting reaction mixture was purged under nitrogen for 20 minutes. 1,2-Dimethylethylenediamine (0.9 g, 10.152 mmol) was added to the resulting reaction mixture under a nitrogen atmosphere. The reaction mixture was heated in a sealed tube at 120 °C for 24 hours. After completion of the reaction, the reaction mixture was filtered through a celite bed and washed with ethyl acetate. The filtrate was diluted with water and extracted with EtOAc, followed by brine. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting crude compound was purified by CombiFlash chromatography (eluting with 30-40% EtOAc in heptane) to give the title compound 1-3 (4.1 g, 41.96%) as an off-white solid. TLC: 50% EtOAc / heptane (R f :0.5). chemical formula C 20 H 17 MS calculated for F2N3OS: 385.11; Found: 385.90 [M+1] + . 1 H NMR (400 MHz, DMSO-d6) δ 7.61 (d, J = 7.8 Hz, 2H), 7.54 - 7.35 (m, 4H), 7.35 - 7.21 (m, 1H), 6.70 (s, 1H), 4.17 (t, J = 5.6 Hz, 2H), 3.81 (t, J = 4.9 Hz, 2H), 2.26 (s, 3H), 2.24 - 2.21 (m, 2H).

[0309] Synthesis of 2-(3-(2',5'-difluoro-[1,1'-biphenyl]-4-yl)-2-oxotetrahydropyrimidin-1(2H)-yl)-4-methylthiazole-5-sulfonic acid (1-4) To a stirred solution of compound 1-3 (4 g, 10.389 mmol) in dry DCM (40 mL) at 0° C. under an inert atmosphere, chlorosulfuric acid (2.07 mL, 31.168 mmol) was added, and the resulting reaction mixture was slowly warmed to room temperature and stirred for 12 hours. After completion of the reaction, the reaction mixture was concentrated to dryness under reduced pressure. The resulting crude residue was purified by trituration with diethyl ether. The resulting solid was filtered and dried in vacuo to give the title compound 1-4 (3.35 g, crude) as an off-white solid. TLC: 100% EtOAc (R f :0.2). chemical formula C 20 H 17 MS calculated for F2N3O4S2: 465.06; Found: 466 [M+1] + .

[0310] Synthesis of 2-(3-(2',5'-difluoro-[1,1'-biphenyl]-4-yl)-2-oxotetrahydropyrimidin-1(2H)-yl)-4-methylthiazole-5-sulfonamide (Compound 1) A stirred solution of compound 1-4 (3.3 g, 7.096 mmol) in POCl (33 mL) was stirred at 90 °C for 5 hours. The reaction mixture was concentrated to dryness under reduced pressure. The resulting residue was dissolved in THF (66 mL), and aqueous ammonia (33 mL) was added at -5 °C, and stirring was continued at room temperature for another 12 hours. After the reaction was completed, the reaction mixture was diluted with water and extracted with EtOAc. The combined organic layers were dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The crude compound was purified by CombiFlash chromatography (eluted with 100% EtOAc) to give the desired product, compound 1 (1.1 g, 44.64%), as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ7.65-7.59 (m, 2H), 7.55 (br s, 2H), 7.53-7.48 (m, 2H), 7.48-7.36 (m, 2H), 7.31-7.25 (m, 1H), 4.17 (t, J = 6.1 Hz, 2H), 3.82 (t, J = 5.6 Hz, 2H), 2.45 (s, 3H), 2.29-2.18 (m, 2H).

[0311] Synthesis of 4'-bromo-2,5-difluoro-1,1'-biphenyl (Intermediate 1A) To a stirred solution of compound 1A (5 g, 17.674 mmol) in 1,4-dioxane:HO (50:5 mL) was added (2,5-difluorophenyl)boronic acid (3.07 g, 19.441 mmol) and KPO (7.5 g, 35.348 mmol), and the reaction mixture was purged under nitrogen for 10 minutes. To the resulting solution was added PdCl(dppf) (1.29 g, 1.767 mmol) under a nitrogen atmosphere. The reaction mixture was heated at 80 °C for 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature, filtered through a pad of Celite, and washed with ethyl acetate. The filtrate was diluted with water and extracted with EtOAc. The combined organic layers were dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The crude compound was purified by CombiFlash chromatography (eluted with 100% heptane) to give the title compound Intermediate 1A (2.3 g, 48.62%) as an off-white solid. TLC: 100% heptane (R f :0.5). 1 H NMR (400 MHz, CDCl3) δ 7.58 (d, J = 8.3 Hz, 2H), 7.40 (d, J = 7.3 Hz, 2H), 7.15-7.06 (m, 2H), 7.05-6.97 (m, 1H).

[0312] Example 9: Biometric Data for 2-(3-(2',5'-difluoro-[1,1'-biphenyl]-4-yl)-2-oxotetrahydropyrimidin-1(2H)-yl)-4-methylthiazole-5-sulfonamide (Compound 1) cell culture Vero cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum and 100 units / mL of penicillin and streptomycin. Cells were passaged 2–3 times per week to maintain subconfluent density.

[0313] HSV-1 antiviral assay Vero cells were plated in a 96-well plate at 2.5 x 10 cells per well. 3 Cells were seeded at a density of 1 / 3 and allowed to attach overnight. After attachment, the medium was replaced with 50 μL of infection medium (DMEM supplemented with 2% fetal bovine serum and 100 units / mL penicillin and streptomycin). Compounds were then added to the cultures in an 8-point, 3-fold serial dilution using a Tecan D300e digital dispenser. DMSO concentrations were normalized to 0.5% for all treatments. After compound addition, 50 μL of infection medium containing 80 TCID50 HSV-1 was added to the cells and incubated at 37°C for 4 days. After incubation, the plates were equilibrated to room temperature, the medium was removed, and a 1:1 dilution of Cell titer glow in phosphate-buffered saline was added to the cells. After a 5-minute incubation, brightness was measured using a Tecan Infinite M1000 Pro plate reader to quantify cell viability.

[0314] HSV-2 antiviral assay Vero cells were plated in a 96-well plate at 1.0 x 10 cells per well. 4 Cells were seeded at a density of 1 / 3 and allowed to attach overnight. After attachment, the medium was replaced with 50 μL of infection medium (DMEM supplemented with 2% fetal bovine serum and 100 units / mL penicillin and streptomycin). Compounds were then added to the cultures in an 8-point, 3-fold serial dilution using a Tecan D300e digital dispenser. DMSO concentrations were normalized to 0.5% for all treatments. After compound addition, 50 μL of infection medium containing 160 TCID50 HSV-2G strain was added to the cells and incubated at 37°C for 5 days. After incubation, 10 μL / well of WST-8 color reagent was added, and the plate was incubated at 37°C for 3 hours. After incubation, cell viability was quantified by measuring absorbance at 460 nm and 620 nm using a Tecan Infinite M1000 Pro plate reader.

[0315] In the HSV-1 antiviral assay, the EC 50The EC value of compound 1 in the HSV-2 antiviral assay was 0.019 μM (n=22). 50 was 0.011 μM (n=35).

[0316] Example 10: Compound 1 PK study in rats Preparation of formulations An aqueous suspension of Compound 1 free form (crystalline form A) in 0.9% NaCl + 0.5% polysorbate 80 vehicle (concentration 3 mg / mL) was prepared as follows: 1) Vehicle (0.9% w / w NaCl and 0.5% w / w polysorbate 80) was prepared by dissolving NaCl and polysorbate 80 in water. 2) An appropriate amount of Compound 1 free form was weighed and added to the vehicle. 3) The mixture was vortexed for 1 minute, stirred for 10 minutes, and then sonicated for 15 minutes. 4) The final formulation was a uniform, cloudy white suspension.

[0317] Formulation characterization The particle size distribution (PSD) of the suspension is Dv 10 =0.57μm, Dv 50 = 2.41 μm, Dv 90 =14.54 μm.

[0318] Animal administration For the pharmacokinetic study, male Sprague Dawley (SD) rats (6-8 weeks old) weighing approximately 230-250 g were used. Animals had free access to food and water. The Compound 1 suspension was administered subcutaneously to each group of animals (n = 3) at a dose of 10 mg / kg in a volume of 3.33 mL / kg.

[0319] Approximately 200 μL of whole blood was collected from the jugular vein into K2EDTA tubes at each time point. The blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. The plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0320] result The mean plasma concentration profiles (n=3) following subcutaneous (SC) administration of the compound suspension at a dose of 10 mg / kg are shown in Figure 10. Long-term exposure of up to at least 58 days was achieved. Plasma concentration variability was low.

[0321] Example 11: Compound 1 PK study in monkeys Preparation of formulations An aqueous suspension of Compound 1 free form (Form A) in 0.9% NaCl + 0.5% Polysorbate 80 vehicle (concentration 3.0 mg / mL) was prepared as follows: 1) Vehicle (0.9% w / w NaCl and 0.5% w / w polysorbate 80) was prepared by dissolving NaCl and polysorbate 80 in water. 2) An appropriate amount of Compound 1 was weighed and added to the vehicle. 3) The mixture was stirred for 15 minutes, then vortexed for 1 minute and sonicated for 10 minutes. 4) The final formulation was a uniform, cloudy white suspension.

[0322] Animal administration For the monkey PK study, male non-naive cyno monkeys weighing approximately 4-6 kg were used. Animals had free access to food and water. The prepared suspension was administered subcutaneously to each animal group (n = 3) at a dose of 10 mg / kg in a volume of 3.33 mL / kg.

[0323] Approximately 0.5 mL of whole blood was collected from the cephalic and saphenous veins at each time point into K2EDTA tubes. Blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. Plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0324] result The mean plasma concentration profiles (n=3) following subcutaneous (SC) administration of Compound 1 suspension are shown in Figure 11. Remarkably, after an initial drug burst, prolonged exposure was achieved over at least 220 days at the administered dose level of 10 mg / kg.

[0325] Example 12: Compound 1 PK study in dogs Preparation of formulations An aqueous suspension of Compound 1 free form (Form A) in 0.9% NaCl + 0.5% Poloxamer 188 vehicle (concentration 5.0 mg / mL) was prepared as follows: 1) Vehicle (0.9% w / w NaCl and 0.5% w / w Poloxamer 188) was prepared by dissolving NaCl and Poloxamer 188 in water. 2) An appropriate amount of Compound 1 free form was weighed and added to the vehicle. 3) The mixture was stirred for 15 minutes, then vortexed for 1 minute and sonicated for 10 minutes. 4) The final formulation was a uniform, cloudy white suspension.

[0326] Animal administration For the canine PK study, male non-naive beagles weighing approximately 10 kg were used. Animals had free access to food and water. The prepared suspension was administered subcutaneously to each group of animals (n = 3) at a dose of 10 mg / kg in a volume of 2 mL / kg.

[0327] Approximately 0.5 mL of whole blood was collected into K2EDTA tubes at each time point. Blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. Plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0328] result The mean plasma concentration profiles (n=3) following subcutaneous (SC) administration of Compound 1 suspension are shown in Figure 12. After an initial drug burst, prolonged exposure was achieved over at least 220 days at the administered dose level of 10 mg / kg.

[0329] Examples 10-12 Summary of PK studies of Compound 1 In three species, a long-acting injectable depot formulation containing Compound 1 was found to be able to provide sustained and stable plasma concentrations of the drug for a significantly extended period after a single subcutaneous administration.

[0330] Additionally, no adverse reactions were observed at the injection site in the test animals.

[0331] Example 13: PK studies after intravenous administration in rats, monkeys and dogs Formulation Preparation - Rat A 0.2 mg / mL solution of Compound 1 in 10% NMP, 10% Solutol HS15, and 80% saline was prepared as follows: 1) A stock solution of Compound 1 in NMP was prepared. 2) Equal volumes of Compound 1 stock solution and Solutol HS15 were mixed together. 3) The above solution was diluted with saline to the target concentration. The final formulation was a clear solution.

[0332] Animal Administration – Rats For the rat pharmacokinetic study, male Sprague Dawley (SD) rats (6-8 weeks old) weighing 230-250 g were used. Animals had free access to food and water. The above Compound 1 solution was administered intravenously to the animals (n=3) at a dose of 1.0 mg / kg.

[0333] Approximately 200 μL of whole blood was collected at each time point into a K2EDTA tube. The blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. The plasma samples were stored at approximately -70°C until analyzed by LC-MS according to the method described above.

[0334] result The mean plasma concentration profiles (n=3) following intravenous (IV) administration of Compound 1 solution at 1.0 mg / kg to rats can be seen in FIG.

[0335] Preparation of Formulations – Monkeys A 0.25 mg / mL solution of Compound 1 in 10% DMSO, 10% Solutol HS15, and 80% saline was prepared as follows: 1) A stock solution of Compound 1 in DMSO was prepared. 2) Equal volumes of Compound 1 stock solution and Solutol HS15 were mixed together. 3) The above solution was diluted with saline to the target concentration. The formulation was a clear solution.

[0336] Animal Administration - Monkeys For the monkey PK study, male non-naive cyno monkeys were used. Animals had free access to food and water. The prepared solution was administered intravenously to each group of animals (n = 3) at a dose of 0.25 mg / kg.

[0337] Approximately 0.5 mL of whole blood was collected into K2EDTA tubes at each time point. Blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. Plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0338] result The mean plasma concentration profiles (n=3) following intravenous (IV) administration of Compound 1 solution at 0.25 mg / kg can be seen in FIG.

[0339] Preparation of Formulations - Dogs A 0.15 mg / mL solution of Compound 1 in 10% DMSO, 10% Solutol HS15, and 80% saline was prepared as follows: 1) A stock solution of Compound 1 in DMSO was prepared. 2) Equal volumes of Compound 1 stock solution and Solutol HS15 were mixed together. 3) The above solution was diluted with saline to the target concentration. The formulation was a clear solution.

[0340] Animal Administration - Dogs For the canine PK study, male non-naive beagles were used. Animals had free access to food and water. The prepared suspension was administered subcutaneously to each group of animals (n = 3) at a dose of 0.15 mg / kg.

[0341] Approximately 0.5 mL of whole blood was collected into K2EDTA tubes at each time point. Blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. Plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0342] result The mean plasma concentration profiles (n=3) following intravenous (IV) administration of Compound 1 solution at 0.15 mg / kg can be seen in FIG.

[0343] summary Compound 1 administered IV to rats, monkeys, and dogs at doses of 1 mg / kg, 0.25 mg / kg, and 0.15 mg / kg, respectively, resulted in long terminal half-lives as shown in the table below:

[0344] [Table 18]

[0345] Example 13A: Further Compound 1 PK studies following IV administration in rats, monkeys, dogs, and minipigs Preparation of formulations – rats A 0.2 mg / mL solution of Compound 1 in 10% NMP, 10% Solutol HS15, and 80% saline was prepared as follows: 1) A stock solution of Compound 1 in NMP was prepared. 2) Equal volumes of Compound 1 stock solution and Solutol HS15 were mixed together. 3) The above solution was diluted with saline to the target concentration. The formulation was a clear solution.

[0346] Animal Administration - Rats For the rat pharmacokinetic study, male Sprague Dawley (SD) rats (6-8 weeks old) weighing 180-184 g were used. Animals had free access to food and water. Compound 1 solution was administered intravenously (1 mL / kg) to each animal (n=3) at a dose of 0.2 mg / kg.

[0347] Approximately 200 μL of whole blood was collected at each time point into a K2EDTA tube. The blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. The plasma samples were stored at approximately -70°C until analyzed by LC-MS according to the method described above.

[0348] Preparation of Formulations – Monkeys A 0.2 mg / mL solution of Compound 1 in 10% DMSO, 10% Solutol HS15, and 80% saline was prepared as follows: 1) A stock solution of Compound 1 in DMSO was prepared. 2) Equal volumes of Compound 1 stock solution and Solutol HS15 were mixed together. 3) The above solution was diluted with saline to the target concentration. The formulation was a clear solution.

[0349] Animal Administration - Monkeys For the monkey PK study, male non-naive Cyno monkeys were used. The animals had free access to food and water. The Compound 1 solution was administered intravenously (1 mL / kg) to each animal (n=3) at a dose of 0.2 mg / kg.

[0350] Approximately 0.5 mL of whole blood was collected into K2EDTA tubes at each time point. Blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. Plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0351] Preparation of Formulations - Dogs A 0.15 mg / mL solution of Compound 1 in 10% DMSO, 10% Solutol HS15, and 80% saline was prepared as follows: 1) A stock solution of Compound 1 in DMSO was prepared. 2) Equal volumes of Compound 1 stock solution and Solutol HS15 were mixed together. 3) The above solution was diluted with saline to the target concentration. The formulation was a clear solution.

[0352] Animal Administration - Dogs For the canine PK study, male non-naive beagles were used. The animals had free access to food and water. The Compound 1 solution was administered intravenously (1 mL / kg) to each animal (n=3) at a dose of 0.15 mg / kg.

[0353] Approximately 0.5 mL of whole blood was collected into K2EDTA tubes at each time point. Blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. Plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0354] Preparation of Formulations – Minipigs A 0.25 mg / mL solution of Compound 1 in 10% DMSO, 10% Solutol HS15, and 80% saline was prepared as follows: 4) A stock solution of Compound 1 in DMSO was prepared. 5) Equal volumes of Compound 1 stock solution and Solutol HS15 were mixed together. 6) The above solution was diluted with saline to the target concentration. The formulation was a clear solution.

[0355] Animal Administration - Miniature Pigs For the minipig PK study, naive Bama pigs (15–16 kg) were used. Animals had free access to food and water. Compound 1 solution was administered intravenously (1 mL / kg) to each animal (n=3) at a dose of 0.25 mg / kg.

[0356] Approximately 0.5 mL of whole blood was collected into K2EDTA tubes at each time point. Blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. Plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0357] result The mean plasma concentration profiles (n=3) following intravenous (IV) injection of Compound 1 solution in rats, dogs, monkeys, and minipigs can be seen in Figure 19(A)-(D).

[0358] summary Compound 1 was administered IV to rats, dogs, monkeys, and minipigs at doses of 0.2 mg / kg, 0.15 mg / kg, 0.2 mg / kg, and 0.25 mg / kg, respectively, resulting in terminal half-lives and clearances as shown in the table below:

[0359] [Table 19]

[0360] Based on the multi-species PK data described above, and using allometric scaling, the human terminal biological half-life of Compound 1 is predicted to be 7.6 days (182 hours), with a predicted clearance of 0.06 L / hour.

[0361] Example 14: Subcutaneous PK study of plitelivir in dogs (A) Preparation of formulations Solution formulation: A solution of pritelivir in 90% NMP (Sigma) and 10% Solutol HS15 (BASF) (concentration 100 mg / mL) was prepared as follows: 1) Approximately 460 mg (based on assay) of crystalline pritelivir free base sesquihydrate was weighed and added to approximately 3.9 mL of NMP. The mixture was stirred for 10 minutes. 2) After the pritelivir was dissolved, Solutol HS15 (approximately 0.4 mL) was added to the solution and mixed for an additional 2 minutes. 3) The final dosage formulation was a clear solution.

[0362] Aqueous microsuspension: An aqueous microsuspension (100 mg / mL concentration) of crystalline plitelivir free base (sesquihydrate) in 2 mg / mL polysorbate 80 (Tween 80) and 5 mg / mL Methocel A4M vehicle was prepared as follows: 1) Crystalline pritelivir free base (sesquihydrate) was jet-milled [milling pressure 0.8 mPa] to produce microparticles (Dv 90 <10 μm). 2) Approximately 460 mg (based on the assay) of the jet-milled material was then dispersed in 2 mg / mL polysorbate 80 (Tween 80) and 5 mg / mL Methocel A4M solution (approximately 4.4 mL) to obtain a 100 mg / mL suspension. 3) The mixture was stirred for 10 minutes. 4) The final formulation was a uniform, cloudy white suspension.

[0363] Formulation characteristics of the microsuspension: The particle size distribution (PSD) of the microsuspension is Dv 10 =1.96μm, Dv 50 = 4.17 μm, and Dv 90 =8.51 μm.

[0364] An SEM image of the microsuspension can be seen in FIG.

[0365] Aqueous nanosuspension: An aqueous nanosuspension (concentration 100 mg / mL) of crystalline plitelivir free base (sesquihydrate) in 10 mg / mL NaCMC, 2 mg / mL polysorbate 80 (Tween 80), and 50 mg / mL mannitol vehicle was prepared as follows: 1) 10 mg / mL crystalline pritelivir free base (sesquihydrate) was dispersed in a solution of 1 mg / mL NaCMC + 0.2 mg / mL polysorbate 80 (Tween 80) and 5 mg / mL mannitol. 2) The suspension was subjected to a high-pressure homogenizer (approximately 20 cycles at 600 bar, followed by 130 cycles at 900 bar) until the particle size was approximately 1 μm. 3) After particle size reduction, the suspension was subjected to freeze-drying to obtain freeze-dried powder. 4) The lyophilized powder was reconstituted with water to give a 100 mg / ml suspension. 5) The suspension was wet milled (10:1 0.1 mm zirconium oxide grinding beads: Priteribil) at 700 rpm for 500 min to reduce the particle size to nano-sized particles (Dv 90 <500nm). 6) The nanosuspension was freeze-dried to obtain a freeze-dried powder. 7) The appropriate amount of lyophilized powder (based on the assay) was weighed out and added to water to obtain a 100 mg / ml suspension. 8) The mixture was stirred for 10 minutes. 9) The final formulation was a uniform, cloudy suspension.

[0366] An SEM image of the nanosuspension can be seen in FIG.

[0367] (B) Administration to animals For the canine PK study, male non-naive beagles were used. The animals had free access to food and water. The prepared formulation was administered subcutaneously to each animal group (n = 3) at a dose of 10 mg / kg in a volume of 0.1 mL / kg.

[0368] Approximately 0.5 mL of whole blood was collected into K2EDTA tubes at each time point. Blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. Plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0369] result The mean plasma concentration profiles (n=3) after subcutaneous (SC) administration of plitelivir formulations are shown in Figure 22. Long-term exposure was achieved at the administered dose level using all three formulations. This solution exhibited the highest plasma concentration levels at early time points. Surprisingly, for the aqueous suspension formulation at early time points (up to 234 hours), higher plasma concentrations of plitelivir were obtained from the microsuspension formulation rather than the nanosuspension formulation. This formulation was able to achieve long-term release using a high concentration of plitelivir, thus allowing administration using a small administration volume of only 0.1 mL / kg. Furthermore, no adverse reactions were observed at the injection site of the test animals.

[0370] Example 15: Further Compound 1 PK studies in rats Preparation of formulations A solution of Compound 1 in 90% NMP and 10% Solutol HS15 (concentration 50 mg / mL) was prepared as follows: 1) An appropriate amount of Compound 1 (Form C) was weighed and added to NMP. 2) After Compound 1 was dissolved, Solutol HS15 was added to the solution and mixed [describe the mixing conditions and time]. 3) The final dosage formulation was a clear solution.

[0371] Animal administration For the pharmacokinetic study, male Sprague Dawley (SD) rats (6-8 weeks old) weighing approximately 230-250 g were used. The animals had free access to food and water. The above Compound 1 solution was administered subcutaneously to each group of animals (n=3) at a dose of 100 mg / kg in a volume of 2 mL / kg.

[0372] At each time point, approximately 150 μL of whole blood was collected from the jugular vein into K2EDTA tubes. Blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. Plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0373] result The mean plasma concentration profiles (n=3) following subcutaneous administration of Compound 1 solution at a dose of 50 mg / kg are shown in Figure 23. After an initial drug burst, prolonged exposure was achieved at the administered dose level for 960 hours (study period). Plasma concentration variability was low. Furthermore, no adverse reactions were observed at the injection site in the test animals.

[0374] Example 16: Further Compound 1 PK studies in dogs Preparation of formulations A solution of Compound 1 in 90% NMP and 10% Solutol HS15 (concentration 50 mg / mL) was prepared as follows: 1) An appropriate amount of crystalline Compound 1 (Form C) was weighed and added to NMP. 2) After Compound 1 was dissolved, Solutol HS15 was added to the solution and mixed. 3) The final dosage formulation was a clear solution.

[0375] A solution of Compound 1 in 90% NMP and 10% Solutol HS15 (concentration 100 mg / mL) was prepared as follows: 1) An appropriate amount of crystalline Compound 1 (Form C) was weighed and added to NMP. 2) After Compound 1 was dissolved, Solutol HS15 was added to the solution and mixed. 3) The final dosage formulation was a clear solution.

[0376] Animal administration For the canine PK study, male non-naive beagles were used. Animals had free access to food and water. The prepared 50 mg / mL solution was administered subcutaneously to each animal group (n=3) at a dose of 10 mg / kg in a volume of 0.2 mL / kg. The prepared 100 mg / mL solution was administered subcutaneously to each animal group (n=3) at a dose of 10 mg / kg in a volume of 0.1 mL / kg.

[0377] Approximately 0.5 mL of whole blood was collected into K2EDTA tubes at each time point. Blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. Plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0378] result The mean plasma concentration profiles (n=3) after subcutaneous administration of Compound 1 solution at two different dose volumes are shown in Figure 24. Using both formulations, long-term exposure of at least 60 days (study period) was achieved at the administered dose level. The formulations were able to achieve long-term release using small dose volumes. Furthermore, no adverse reactions were observed at the injection site of the test animals.

[0379] Example 17: Subcutaneous and intramuscular PK study of Compound 1 in rats using microsuspension and nanosuspension formulations (A) Preparation of formulations Aqueous microsuspension: An aqueous microsuspension (concentration 200 mg / mL) of crystalline Compound 1 free form (crystalline Form C) in 20 mg / mL PVP K30, 10 mg / mL poloxamer 188, and 50 mg / mL mannitol vehicle was prepared as follows: 1) Crystalline Compound 1 (Form C) was jet-milled to produce microparticles (Dv 90 <10 μm). 2) The jet milled material was dispersed in 20mg / ml PVP K30 + 10mg / ml Poloxamer 188 + 50mg / ml Mannitol to obtain a uniform suspension. 3) The suspension was freeze-dried to obtain a freeze-dried powder. 4) The appropriate amount of lyophilized powder (based on the assay) was weighed and added to water to obtain a 200 mg / ml suspension. 5) The mixture was stirred for 5 minutes. 6) The final formulation was a uniform, cloudy white suspension.

[0380] Formulation characteristics of the microsuspension: The particle size distribution (PSD) of the microsuspension is Dv10 =1.19μm, Dv 50 = 2.76 μm, and Dv 90 =5.65 μm.

[0381] An SEM image of the microsuspension can be seen in FIG.

[0382] Aqueous nanosuspension: An aqueous nanosuspension (concentration 200 mg / mL) of crystalline Compound 1 free form (crystalline Form C) in 20 mg / mL PVP K30, 10 mg / mL poloxamer 188, and 50 mg / mL mannitol vehicle was prepared as follows: 1) 20 mg / mL crystalline Compound 1 (Form C) was dispersed in a solution of 2 mg / mL PVP K30, 1.0 mg / mL poloxamer 188, and 5 mg / mL mannitol. 2) The suspension was subjected to a high-pressure homogenizer until the particle size reached approximately 1 μm. 3) After particle size reduction, the suspension was freeze-dried to obtain a freeze-dried powder. 4) The lyophilized powder was reconstituted with water to give a 100 mg / mL suspension. 5) The suspension was wet milled (10:1 0.1 mm zirconium oxide grinding beads: Compound 1) at 900 rpm for 600 minutes to reduce the particle size to nano-sized particles (Dv 90 <200nm). 6) The nanosuspension was freeze-dried to obtain a freeze-dried powder. 7) Weigh out the appropriate amount of lyophilized powder (based on assay) and add water to obtain a 200 mg / ml suspension. 8) The mixture was stirred for 10 minutes. 9) The final formulation was a uniform, cloudy suspension.

[0383] Formulation characteristics of nanosuspension: The particle size distribution (PSD) of the nanosuspension is Dv 10 =69.1nm, Dv 50 = 77.7 nm, and Dv 90 =88.5nm.

[0384] An SEM image of the nanosuspension can be seen in FIG.

[0385] (B) Administration to animals For the pharmacokinetic study, male Sprague Dawley (SD) rats (6-8 weeks old) weighing approximately 230-250 g were used. Animals had free access to food and water. The above Compound 1 solution was administered subcutaneously or intramuscularly to each animal group (n = 3) at a dose of 20 mg / animal (approximately 80 mg / kg) in a volume of 0.1 mL / animal (approximately 0.4 mL / kg).

[0386] At each time point, approximately 150 μL of whole blood was collected from the jugular vein into K2EDTA tubes. Blood samples were placed on ice and centrifuged at 2000 g for 5 minutes to obtain plasma samples within 15 minutes. Plasma samples were stored at approximately -70°C until analysis by LC-MS.

[0387] result The mean plasma concentration profiles (n=3) after subcutaneous administration of Compound 1 microsuspension and Compound 1 nanosuspension can be seen in Figure 27. Both formulations were able to provide long-term exposure of at least 600 hours (the test period). The nanosuspension provided higher plasma levels than the microsuspension during the test period. The formulations were able to achieve long-term release using high drug loading and small administration volumes. Furthermore, no adverse reactions were observed at the injection site in the test animals.

[0388] The mean plasma concentration profiles (n=3) after intramuscular administration of Compound 1 microsuspension and Compound 1 nanosuspension can be seen in Figure 28. Both formulations were able to provide prolonged exposure of at least 600 hours (the duration of the study), and plasma levels were higher than those achieved with these formulations when administered by subcutaneous injection. The nanosuspension provided higher plasma levels than the microsuspension. The formulations were able to achieve prolonged release using high drug loading and small administration volumes. Furthermore, no side effects were observed at the injection site in the test animals.

Claims

1. a. Helicase / primase inhibitors or pharmaceutically acceptable salts thereof, b. A long-acting injectable depot pharmaceutical composition comprising one or more pharmaceutically acceptable excipients, wherein the helicase / primase inhibitor has a human biological terminal half-life of 10 hours or more.

2. The composition according to claim 1, wherein the composition is a long-acting injectable depot pharmaceutical composition for subcutaneous or intramuscular use.

3. The composition according to claim 2, wherein the composition is for administration twice a month, once a month, once every two months, once every three months, once every six months, or once a year.

4. In vitro EC with helicase primase inhibitors at a concentration of less than 0.1 μM against HSV-1 and / or HSV-2 50 The composition according to claim 1, having a value.

5. Helicase primase inhibitors include N-methyl-N-(4-methyl-5-sulfamoyl-1,3-thiazole-2-yl)-2-[4-(pyridine-2-yl)phenyl]acetamide: 【Chemistry 1】 The composition according to claim 1, or a pharmaceutically acceptable salt thereof.

6. The composition according to claim 1, wherein the helicase primase inhibitor is N-methyl-N-(4-methyl-5-sulfamoyl-1,3-thiazole-2-yl)-2-[4-(pyridine-2-yl)phenyl]acetamide free base.

7. The composition according to claim 6, wherein the free base exists as a crystalline solid.

8. The composition according to claim 6, wherein the long-acting injectable depot pharmaceutical composition is an aqueous suspension in which a free base is suspended as a crystalline solid.

9. The composition according to claim 8, wherein the long-acting injectable depot pharmaceutical composition is an aqueous nanosuspension in which a free base is suspended as a crystalline solid.

10. The composition according to claim 1, wherein the long-acting injectable depot pharmaceutical composition comprises an organic solvent solution formulation.

11. The composition according to claim 10, wherein the organic solvent is selected from benzyl alcohol, benzyl benzoate, N-methylpyrrolidone (NMP), 2-pyrrolidone, ethanol, propylene glycol, acetone, methyl acetate, ethyl acetate, methyl ethyl ketone, dimethylacetamide (DMA), dimethylformamide, dimethyl sulfoxide (DMSO), tetrahydrofuran, caprolactam, oleic acid, 1-dodecyl azacycloheptan-2-one, and mixtures thereof.

12. The composition according to claim 10, wherein the helicase primase inhibitor is N-methyl-N-(4-methyl-5-sulfamoyl-1,3-thiazole-2-yl)-2-[4-(pyridine-2-yl)phenyl]acetamide, and the organic solvent is selected from NMP, DMA, and DMSO.

13. Helicase primase inhibitor 2-(3-(2',5'-difluoro-[1,1'-biphenyl]-4-yl)-2-oxotetrahydropyrimidine-1(2H)-yl)-4-methyl-thiazole-5-sulfonamide (compound 1): 【Chemistry 2】 The composition according to claim 1, or a pharmaceutically acceptable salt thereof.

14. The composition according to claim 1, wherein the helicase primase inhibitor is the free form of 2-(3-(2',5'-difluoro-[1,1'-biphenyl]-4-yl)-2-oxotetrahydropyrimidine-1(2H)-yl)-4-methylthiazole-5-sulfonamide.

15. The composition according to claim 14, wherein the free form exists as a crystalline solid.

16. The composition according to claim 14, wherein the long-acting injectable depot pharmaceutical composition is an aqueous suspension, and the free form is suspended as a crystalline solid.

17. The composition according to claim 16, wherein the long-acting injectable depot pharmaceutical composition is an aqueous microsuspension or aqueous nanosuspension, and the free form is suspended as a crystalline solid.

18. The composition according to claim 13, wherein the long-acting injectable depot pharmaceutical composition comprises an organic solvent solution formulation.

19. The composition according to claim 18, wherein the organic solvent is selected from N-methylpyrrolidone (NMP), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), benzyl alcohol, benzyl benzoate, and mixtures thereof.

20. The composition according to claim 19, wherein the organic solvent is selected from NMP, DMA, and DMSO.

21. The composition according to claim 1, wherein the long-acting injectable depot pharmaceutical composition contains about 2 wt% to about 40 wt% of a helicase primase inhibitor or a pharmaceutically acceptable salt thereof.

22. The composition according to claim 1, wherein the long-acting injectable depot pharmaceutical composition comprises about 100 mg to about 1200 mg, about 10 mg to about 900 mg, or about 100 mg to about 600 mg of a helicase primase inhibitor or a pharmaceutically acceptable salt thereof.

23. The composition according to claim 1, wherein the long-acting injectable depot pharmaceutical composition contains N-methyl-N-(4-methyl-5-sulfamoyl-1,3-thiazole-2-yl)-2-[4-(pyridine-2-yl)phenyl]acetamide free base as a helicase primase inhibitor, and the composition results in a plasma concentration of N-methyl-N-(4-methyl-5-sulfamoyl-1,3-thiazole-2-yl)-2-[4-(pyridine-2-yl)phenyl]acetamide in a subject after administration of at least 25 ng / mL for at least 80% of the administration interval, and the administration interval is at least 10 days.

24. The composition according to claim 1, wherein the long-acting injectable depot pharmaceutical composition comprises the free form of 2-(3-(2',5'-difluoro-[1,1'-biphenyl]-4-yl)-2-oxotetrahydropyrimidine-1(2H)-yl)-4-methylthiazole-5-sulfonamide as a helicase primase inhibitor, the composition yields a plasma concentration of 2-(3-(2',5'-difluoro-[1,1'-biphenyl]-4-yl)-2-oxotetrahydropyrimidine-1(2H)-yl)-4-methylthiazole-5-sulfonamide in a subject after administration of at least 25 ng / mL for at least 80% of the administration interval, and the administration interval is at least 10 days.

25. The composition according to claim 1, wherein the composition is stable for at least four weeks.

26. A composition according to any one of claims 1 to 25 for use as a pharmaceutical.

27. A pharmaceutical composition according to any one of claims 1 to 25 for treating a herpesvirus (e.g., HSV) infection in a subject requiring treatment, characterized in that the pharmaceutical composition is administered to the subject in a therapeutically effective amount of the helicase primase inhibitor or a pharmaceutically acceptable salt thereof.

28. The composition according to claim 27, wherein the pharmaceutical composition is administered twice a month, once a month, once every two months, once every three months, once every six months, or once a year.

29. The composition according to claim 27, wherein the pharmaceutical composition is administered once a month or once every three months.

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