DESC:TECHNICAL FILED
[0001] The present disclosure pertains to technical field of therapeutics and, more particularly to pharmaceutical compositions and methods of treatment of endometriosis in humans.

BACKGROUND
[0002] Endometriosis is a benign condition in which ectopic endometrial glands and stroma are present outside of the uterine cavity. This condition is sometimes denoted as endometriosis externa or adenomyosis externa. The endometriotic tissues possess receptors for estrogen and progestogen, allowing them to proliferate and differentiate in response to hormonal fluctuations throughout the menstrual cycle. Typically, endometriosis is localized to the peritoneal or serosal surfaces of abdominal organs, notably the ovaries, posterior broad ligament, posterior cul-de-sac, and uterosacral ligaments, with the potential formation of uterosacral nodules. Less common sites encompass the serosal surfaces of the small and large bowel, ureters, bladder, vagina, surgical scars, pleura, and pericardium. Clinical indications of endometriosis comprise pelvic pain, the presence of a pelvic mass, irregularities in menstrual patterns, and infertility. Lesions affecting the bowel or bladder can induce pain during defecation or urination, abdominal distension, and rectal bleeding during menses (as many endometriotic implants are prone to bleeding during menstruation). On the ovaries or adnexal structures, these implants can result in the formation of an endometrioma (a cystic mass localized within an ovary) or adnexal adhesions. It has been reported that endometriosis is present in 10-15% of menstruating women aged 15 to 44 and in 25-50% of women experiencing infertility.
[0003] Existing treatment options for endometriosis include androgenic hormonal drugs such as Danazol. Danazol is supplied as 50, 100, and 200 mg tablets, and the drug is administered orally. For mild endometriosis, two 100 or 200 mg tablets twice daily, and for moderate to severe endometriosis, 400 mg tablets twice daily are recommended, but the dose should be titrated down to the minimum required to maintain amenorrhea. Therapy typically lasts six months but can be extended to 9 months. As a synthetic steroid, danazol has many adverse effects in line with other androgens, making its adverse effects broad and systemic in nature. Also, since this drug is a synthetic steroid, elevated blood pressure can occur, along with its effects on adrenal activity. The most commonly reported side effects of danazol include weight gain, gastrointestinal symptoms including bloating, nausea, vomiting, gastroenteritis, elevated liver function, joint pain, muscle spasm, lethargy, headache, depression, atrophic vaginitis and adverse effects on lipids. Other than these, gynaecologic side effects such as intermenstrual bleeding, breast atrophy, flushes, and androgenic side effects such as hirsutism, decreased breast size, acne, hair loss, oily skin, oily hair, menstrual irregularities and hoarseness, side effects due to weak mineralocorticoid activity such as swelling and edema have also been reported.
[0004] Gonadotrophin-releasing hormone analogues (GnRHas) for pain associated with endometriosis: ? GnRHas are a group of drugs often used to treat endometriosis by decreasing hormone levels. Clinical studies suggested that treatment with a GnRHa improved symptom relief compared with no treatment or placebo. There was no evidence of improvement when compared with danazol or intra-uterine progestogen (levonorgestrel). However, there were more side effects in the GnRHa group compared with the danazol group. There was not enough evidence to make clear if higher or lower doses of GnRHa were better, or which length of treatment was best.
[0005] Gonadotrophin-releasing hormone analogues (GnRHas) for endometriosis: bone mineral density: ? Possible adverse effects of GnRHas include loss of bone density, loss of sex-drive, vaginal dryness and hot flushes. The decrease in bone density is particularly important because it increases the risk of osteoporosis so the treatment is usually limited to less than six months.
[0006] Progestagens and anti-progestagens for pain associated with endometriosis: ? Progestagens and anti-progestagens are some of the hormonal drugs used for treatment of pain in endometriosis. Prior art has limited evidence for the effectiveness of these drugs in the reduction of pain in women with endometriosis. There was no evidence of a benefit of depot or oral progestagens over other treatment. There was no evidence of a benefit of anti-progestagens.
[0007] Progesterone receptor modulators for endometriosis: ? Progesterone receptor modulators have been advocated as one of the hormonal treatments for endometriosis. Mifepristone may relieve dysmenorrhoea (painful periods) and dyspareunia (pain during sexual intercourse) compared to placebo in women with endometriosis. However, amenorrhoea (absence of menstrual periods) and hot flushes were common side effects of mifepristone.
[0008] Further medical therapies for endometriosis involve estrogen suppression, such as by administration of continuous oral contraception with estrogen/progestogen combination products. Usual side effects of these therapies include abdominal swelling, breast tenderness, breakthrough bleeding, and deep vein thrombosis.
[0009] Despite the existence of multiple treatments for endometriosis, several studies suggest that none of these treatments are as effective as desired. Accordingly, there is a need to develop effective medical treatments for endometriosis, including endometriosis externa, endometrioma, adenomyosis, adenomyomas, endometriotic or adenomyotic nodules of the uterosacral ligaments and endometriotic nodules such as scar endometriosis. The present disclosure satisfies these needs and provides further related advantages.

SUMMARY
[00010] Aspects of the present disclosure relate to the use of thymosin alpha-1 for the treatment of endometriosis and symptoms thereof in a subject. The present disclosure is based at least in part on the clinical observation that thymosin alpha-1 is effective in the treatment of endometriosis, including endometriosis externa, endometrioma, adenomyosis, adenomyomas, adenomyotic nodules of the uterosacral ligaments, and endometriotic nodules, such as scar endometriosis.
[00011] Accordingly, in one aspect, the present disclosure is directed to a method for the treatment of endometriosis in a subject in need thereof by administering a therapeutically effective amount of thymosin alpha-1.
[00012] The therapeutically effective amount of thymosin alpha-1 generally ranges from 0.001 mg/kg/day (mass of drug compared to mass of subject) to 100 mg/kg/day, preferably from 0.01 mg/kg/day to 10 mg/kg/day. In some embodiments, administering a therapeutically effective amount comprises administering a flat dose of 0.1-50 mg of thymosin alpha-1 to the subject, over the course of treatment. In one embodiment, the course of treatment is from about 3 to about 12 months or longer. In some embodiments, the course of treatment is from about 3 to about 9 months. In some embodiments, the course of treatment is from about 6 to about 12 months or longer.
[00013] Another aspect of this disclosure provides a pharmaceutical composition for use in the treatment of endometriosis, which comprises therapeutic amounts of thymosin alpha-1 and one or more pharmaceutically acceptable excipients.
[00014] In some embodiments, the pharmaceutical composition comprises: about 0.1 mg/ml to about 50 mg/ml of thymosin alpha-1; about 1% to about 8% weight/volume (w/v) stabilizer; and about 1 mM to about 20 mM buffer.
[00015] Further provided, in one aspect, is the use of thymosin alpha-1 in the manufacture of a medicament for use in the treatment of endometriosis.
[00016] According to embodiments of the present disclosure, thymosin alpha-1 or the pharmaceutical composition provided herein can be administered alone or in combination with a further co-agent useful in the treatment of endometriosis, e.g., in combination with antiandrogens, anti-progestins or progestins. The combination of thymosin alpha-1 and the co-agent can be administered together in a single composition or administered separately in two or more different compositions. The administration of the thymosin alpha-1 and the co-agent can be in any order. For example, the thymosin alpha-1 can be administered concurrently with, prior to, or subsequent to, the co-agent.
[00017] Various objects, features, aspects and advantages of the inventive subject matter will become more apparent from the following detailed description of preferred embodiments.

DETAILED DESCRIPTION
[00018] Thymosin alpha-1, a biologically active peptide consisting of 28 amino acid residues, was first described and characterized by Goldstein et al. in Proceedings of the National Academy of Sciences of the USA 1977;74(2):725–729. Thymosin alpha 1 is an acetylated polypeptide with the following sequence: Ac-Ser-Asp-Ala-Ala-Val-Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu-Lys-Glu-Lys-Lys-Glu-Val-Val-Glu-Glu-Ala-Glu-Asn-OH, and having a molecular weight of 3108 Daltons. Thymosin alpha 1 has long been recognized as an immune enhancing, immune modulating, as well as an immune restoring agent, and as such it has been utilized in several clinical and research settings. Thymalfasin is the synthetic form of thymosin alpha-1.
[00019] “Endometriosis”, as used herein, refers to any non-malignant disorder in which functioning endometrial tissue is present in a location in the body other than the endometrium of the uterus, i.e. outside the uterine cavity or is present within the myometrium of the uterus. For purposes herein it also includes conditions, such as adenomyosis/adenomyoma, that exhibit myometrial tissue in the lesions. Thus, the term “endometriosis” includes “endometriosis” as defined in The Merck Manual, where the endometrial tissue is present outside the uterine cavity, including uterosacral nodules, endometriomas, adnexal adhesions, and adenomyosis, where the endometrial tissue is present within the myometrium of the uterus.
[00020] Endometriosis, as used herein, thus includes the conditions commonly referred to as endometriosis externa (or endometriosis as defined in The Merck Manual), endometrioma, adenomyosis, adenomyoma, endometriotic or adenomyotic nodules of the uterosacral ligaments, endometriotic nodules elsewhere such as scar endometriosis, and any nonmalignant disorder in which functioning endometrial tissue is present at a locus other than the endometrium.
[00021] As used herein, “endometriotic tissue” is endometrial tissue seen in endometriosis, that is, the endometrial tissue present in a location other than the endometrium of the uterus. Myometrial tissue refers to tissue in the muscle layer of the uterus. This tissue also occurs in lesion treated herein.
[00022] The terms “treat”, “treating” and “treatment” include one or more of: reducing the frequency and/or severity of symptoms, elimination of symptoms and/or their underlying cause, and improvement or remediation of damage. Thus, treatment of endometriosis includes, for example, relieving the pain experienced by a woman suffering from endometriosis, and/or causing the regression or disappearance of endometriotic lesions.
[00023] As used herein, a “subject” includes any mammals, typically female mammals, including humans, for whom treatment is contemplated. Subjects also are referred to as patients.
[00024] Unless otherwise noted, the term “about”, when modifying the quantity (e.g., mM, or M) of an ingredient or composition, the percentage (v/v or w/v) of a formulation component, or the pH of a solution/formulation, or the like refers to variation in the numerical quantity that can occur. In certain embodiments, “about” can mean a variation of ± 0.1%, 0.5%, 1%, 2%, or 3%.
[00025] As used herein, an “effective amount” or a “therapeutically effective amount” of thymosin alpha-1 of means a sufficient amount to effect “treatment” as defined. Treatment can be associated with undesirable effects (“side effects”) along with the desired therapeutic effect, so that a medical practitioner prescribing or performing treatment will balance the potential benefits against the potential risks in determining what constitutes an appropriate “effective amount” or “therapeutically effective amount”. Also, because the quantity of endometriotic tissue will vary from woman to woman, the “effective amount” of thymosin alpha-1 to be administered can vary. Thus it is not possible to specify an exact “effective amount”; In view of the disclosure herein, however, the skilled medical practitioner, can determine an appropriate “effective amount” in any individual case.
[00026] The term thymosin alpha 1 (T?1) as used herein encompasses not only native (i.e., naturally occurring) T?1 but also synthetic T?1, and recombinant T?1 having the amino acid sequence of native T?1, amino acid sequences substantially similar thereto, or an abbreviated sequence from thereof, and their biologically active analogs (including muteins) having substituted, deleted, elongated, replaced, or otherwise modified sequences which possess bioactivity substantially similar to that of T?1.
[00027] The present disclosure is based at least in part on the clinical observation that thymosin alpha-1 is effective in the treatment of endometriosis, including endometriosis externa, endometrioma, adenomyosis, adenomyomas, adenomyotic nodules of the uterosacral ligaments, and endometriotic nodules, such as scar endometriosis.
[00028] Accordingly, one aspect of the present disclosure is directed to a method for the treatment of endometriosis in a subject in need thereof by administering to the subject a therapeutically effective amount of thymosin alpha-1.
[00029] The therapeutically effective amount of thymosin alpha-1 generally ranges from 0.001 mg/kg/day (mass of drug compared to mass of subject) to 100 mg/kg/day, preferably from 0.01 mg/kg/day to 10 mg/kg/day. In some embodiments, administering a therapeutically effective amount comprises administering a flat dose of 0.1-50 mg of thymosin alpha-1 to the subject. In some embodiments, the therapeutically effective amount of thymosin alpha-1 is from about 0.1 mg to about 50 mg per day. In some embodiments, the therapeutically effective amount of thymosin alpha-1 is from about 0.5 mg to about 10 mg per day. In one preferred embodiment, the therapeutically effective amount of thymosin alpha-1 is from about 1 mg to about 4 mg per day.
[00030] According to embodiments of the present disclosure, thymosin alpha-1 can be administered to the subject as a course of treatment, e.g., administration at various time intervals, such as once per day over the course of treatment, once every two days over the course of treatment, once every three days over the course of treatment, once every four days over the course of treatment, once every five days over the course of treatment, once every six days over the course of treatment, once per week over the course of treatment, twice weekly over the course of treatment, thrice weekly over the course of treatment, once every other week over the course of treatment, once per month over the course of treatment, etc. Time courses can vary, and can continue as long as desired effects are obtained. In one embodiment, the course of treatment is from about 3 to about 12 months or longer. In some embodiments, the course of treatment is from about 3 to about 9 months. In some embodiments, the course of treatment is from about 6 to about 12 months or longer.
[00031] In some embodiments, thymosin alpha-1 is administered to a subject by subcutaneous injection twice weekly in a pharmaceutical composition within the range of about 1-4 mg, e.g., about 1.6 mg. Such administration preferably occurs for at least about 6 months, more preferably, for about 3 to 12 months. The composition comprising thymosin alpha-1 can be a reconstituted liquid composition, or it can be a liquid composition not previously lyophilized.
[00032] In certain embodiments, the subject is a human subject. In one embodiment, the subject is a female human subject.
[00033] The present disclosure also provides a pharmaceutical composition for use in the treatment of endometriosis, which comprises therapeutic amounts of thymosin alpha-1 and one or more of pharmaceutically acceptable excipients.
[00034] In various embodiments, thymosin alpha-1 can be formulated with one or more pharmaceutically acceptable excipients to produce a composition which is suitable for oral administration or parenteral administration.
[00035] In some embodiments, the composition described herein is in aqueous solution. In alternative embodiments, the present disclosure provides lyophilized compositions made by lyophilizing an aqueous formulation comprising thymosin alpha-1.
[00036] In one embodiment, the pharmaceutical composition is a liquid.
[00037] In one embodiment, the pharmaceutical composition is a reconstituted solution from a lyophilized composition.
[00038] In one embodiment, the pharmaceutical composition is a reconstituted solution from a lyophilized composition, which is useful for subcutaneous delivery to a human patient in need thereof.
[00039] In various embodiments, thymosin alpha-1 or the pharmaceutical composition provided herein can be administered alone or in combination with a further co-agent useful in the treatment of endometriosis, e.g., in combination with antiandrogens, anti-progestins or progestins. The combination of thymosin alpha-1 and the co-agent can be administered together in a single composition or administered separately in two or more different compositions. The administration of the thymosin alpha-1 and the co-agent can be in any order. For example, the thymosin alpha-1 can be administered concurrently with, prior to, or subsequent to, the co-agent.
[00040] In some embodiments, the pharmaceutical composition disclosed herein is formulated for parenteral administration, such as, for example, intravenous, intramuscular, or subcutaneous injection. Such parenteral formulations may comprise one or more pharmaceutically acceptable excipients, such as buffer, stabilizer, surfactant, anti-oxidant, diluent, and the like.
[00041] In one preferred embodiment, the present disclosure provides a pharmaceutical composition for treating endometriosis, which comprises: about 0.1 mg/ml to about 50 mg/ml of thymosin alpha-1; about 1% to about 8% weight/volume (w/v) stabilizer; and about 1 mM to about 20 mM buffer.
[00042] In various embodiments, the buffer provides the composition a pH in the range from about 6.0 to about 7.5. In embodiments where the composition is lyophilized, the pH refers to the pH after reconstitution of the lyophilized composition of this disclosure. Examples of buffers that will control the pH in this range include succinate (sodium or potassium), histidine, sodium acetate, phosphate (sodium or potassium), Tris (tris(hydroxymethyl) aminomethane), diethanolamine, citrate (sodium) and other organic acid buffers. Buffer(s) can be used in the range from about 1 mM to about 20 mM.
[00043] In one embodiment, the buffer is phosphate buffer.
[00044] In one particularly preferred embodiment, the buffer is combination of disodium hydrogen phosphate and monobasic sodium phosphate.
[00045] The stabilizer can be selected from sucrose, trehalose, (2-hydroxypropyl)-ß-cyclodextrin, mannitol, sorbitol, L-arginine, a pharmaceutically acceptable salt of L-arginine, L-proline, a pharmaceutically acceptable salt of L-proline, glycine, a pharmaceutically acceptable salt of glycine, and a combination thereof. Preferably, the stabilizer is present in an amount of from about 1% to about 8% weight/volume (w/v) based on the total amount of the composition. In one embodiment, the stabilizer is present in an amount of from about 1mg to about 100 mg per vial.
[00046] In one particularly preferred embodiment, the stabilizer is mannitol.
[00047] In certain embodiments of the present disclosure, thymosin alpha-1 is formulated as a lyophilized powder for reconstituting and utilizing for subcutaneous administration. The lyophilized compositions are reconstituted prior to administration. The thymosin alpha-1 may be reconstituted at a concentration ranging from about 0.1 mg/ml to about 50 mg/ml. In some embodiments, thymosin alpha-1 may be reconstituted at a concentration in the range of from about 0.5 mg/ml to about 10 mg/ml. In one particularly preferred embodiment, thymosin alpha-1 is reconstituted at a concentration of 1.6 mg/ml. In certain embodiments, thymosin alpha-1 is provided at about 1.6 mg/vial, and is reconstituted with sterile water for injection prior to use. In some embodiments, the pH of the reconstituted solution is 6.0 to 7.5.
[00048] The lyophilized compositions of this disclosure are formed by lyophilization (freeze-drying) of a pre-lyophilization solution. The pre-lyophilization solution may comprise thymosin alpha-1 at a concentration of from about 0.1 mg/ml to about 50 mg/ml. Such pre-lyophilization solutions may be at pH 6.0 to 7.5. Freeze-drying may be accomplished by any method known in the art. Freeze-drying conditions can be varied depending on the formulation and vial size. In some embodiments, it may be desirable to lyophilize the pre-lyophilization solution in the container in which reconstitution of thymosin alpha-1 is to be carried out, in order to avoid a transfer step.
[00049] The compositions disclosed herein may be presented in unit-dose or multi-dose containers, for example, sealed ampules and vials, and may be stored in a freeze-dried (lyophilized) form requiring only the addition of sterile liquid carrier, for example water for injection, immediately prior to use.
[00050] In one preferred embodiment, the present disclosure provides a liquid thymosin alpha-1 composition that is reconstituted from a lyophilized composition wherein the reconstituted solution comprises: about 0.1 mg/ml to about 50 mg/ml of thymosin alpha-1, about 1% to about 8% weight/volume (w/v) stabilizer, and about 1 mM to about 20 mM buffer.
[00051] In one preferred embodiment, the present disclosure provides a liquid thymosin alpha-1 composition that is reconstituted from a lyophilized composition wherein the reconstituted solution comprises: about 0.1 mg/ml to about 50 mg/ml of thymosin alpha-1, about 1mg/ml to about 100 mg/ml stabilizer, and about 1 mM to about 20 mM buffer.
[00052] In one particularly preferred embodiment, the present disclosure provides a liquid thymosin alpha-1 composition that is reconstituted from a lyophilized composition wherein the reconstituted solution comprises: 1.6 mg/ml of thymosin alpha-1, 50 mg/ml of mannitol, 1.059 mg/ml of disodium hydrogen phosphate and 0.56 mg/ml of monobasic sodium phosphate.
[00053] The present disclosure also provides a method for treating endometriosis in a subject, the method comprising administering an effective amount of the composition of this disclosure to the subject. In certain embodiments, the composition is administered to the subject by subcutaneous administration. Subcutaneous administration may be performed by using a syringe, or using other injection devices, injector pens or needleless devices.
[00054] In some embodiments of the method, the effective amount comprises a dose of thymosin alpha-1 in the range of from 0.01 mg/kg subject body weight per day to 10 mg/kg subject body weight per day. In some embodiments of the method, the effective amount comprises a dose of thymosin alpha-1 in the range of from 0.5 mg to 10 mg per day. In some embodiments of the method, the thymosin alpha-1 composition is administered by subcutaneous administration.
[00055] In some embodiments, the method can be used to treat endometriosis, including endometriosis externa, endometrioma, adenomyosis, adenomyomas, adenomyotic nodules of the uterosacral ligaments, and/or endometriotic nodules.
[00056] Another aspect of the present disclosure relates to the use of thymosin alpha-1 in the manufacture of a medicament for use in the treatment of endometriosis in a subject.
[00057] The following list of embodiments forms part of the description.
[00058] Embodiment 1: A pharmaceutical composition for treating endometriosis, comprising:
about 0.1 mg/ml to about 50 mg/ml of thymosin alpha-1;
about 1% to about 8% weight/volume (w/v) stabilizer; and
about 1 mM to about 20 mM buffer.
[00059] Embodiment 2: The pharmaceutical composition of one or more of Embodiments 1-9, wherein the composition has a pH between 6.0 and 7.5.
[00060] Embodiment 3: The pharmaceutical composition of one or more of Embodiments 1-9, wherein the stabilizer is selected from the group consisting of sucrose, trehalose, (2-hydroxypropyl)-ß-cyclodextrin, mannitol, sorbitol, L-arginine, a pharmaceutically acceptable salt of L-arginine, L-proline, a pharmaceutically acceptable salt of L-proline, glycine, a pharmaceutically acceptable salt of glycine, and a combination thereof.
[00061] Embodiment 4: The pharmaceutical composition of one or more of Embodiments 1-9, wherein the buffer is selected from the group consisting of a succinate buffer, a histidine buffer, a sodium acetate buffer, a phosphate buffer, tris (tris(hydroxymethyl) aminomethane), diethanolamine, a citrate buffer, and a combination thereof.
[00062] Embodiment 5: The pharmaceutical composition of one or more of Embodiments 1-9, wherein the buffer comprises disodium hydrogen phosphate and monobasic sodium phosphate.
[00063] Embodiment 6: The pharmaceutical composition of one or more of Embodiments 1-9, wherein the stabilizer is mannitol and the buffer comprises disodium hydrogen phosphate and monobasic sodium phosphate, the composition comprising:
about 0.5 mg/ml to about 25 mg/ml of thymosin alpha-1;
about 2% to about 7% w/v mannitol;
about 4 mM to about 10 mM disodium hydrogen phosphate buffer; and
about 1 mM to about 8 mM monobasic sodium phosphate buffer.
[00064] Embodiment 7: The pharmaceutical composition of one or more of Embodiments 1-9, comprising:
about 1.6 mg/ml of thymosin alpha-1;
about 5% w/v mannitol;
about 7.5 mM disodium hydrogen phosphate buffer; and
about 4.5 mM monobasic sodium phosphate buffer.
[00065] Embodiment 8: The pharmaceutical composition of one or more of Embodiments 1-9, wherein the composition is a liquid.
[00066] Embodiment 9: The pharmaceutical composition of one or more of Embodiments 1-9, wherein the composition is a reconstituted solution from a lyophilized composition.
[00067] Embodiment 10: A liquid composition that is reconstituted from a lyophilized composition wherein the reconstituted solution comprises:
1.6 mg/ml of thymosin alpha-1;
50 mg/ml of mannitol;
1.059 mg/ml of disodium hydrogen phosphate; and
0.56 g/ml of monobasic sodium phosphate.
[00068] Embodiment 11: A method of treating endometriosis in a subject in need thereof, the method comprising administering an effective amount of thymosin alpha-1 to the subject.
[00069] Embodiment 12: The method of one or more of Embodiments 11-13, wherein the effective amount comprises a dose of thymosin alpha-1 ranging from 0.01 mg/kg/day to 10 mg/kg/day; or wherein the effective amount comprises a dose of thymosin alpha-1 ranging from 0.5 mg to 10 mg per day; or wherein thymosin alpha-1 is administered by subcutaneous administration.
[00070] Embodiment 13: The method of one or more of Embodiments 11-13, wherein the endometriosis is endometriosis externa, endometrioma, adenomyosis, adenomyomas, adenomyotic nodules of the uterosacral ligaments, and/or endometriotic nodules.
[00071] Embodiment 14: Use of thymosin alpha-1 in the manufacture of a medicament for use in the treatment of endometriosis in a subject.
[00072] Embodiment 15: The use of Embodiment 14, wherein the endometriosis is endometriosis externa, endometrioma, adenomyosis, adenomyomas, adenomyotic nodules of the uterosacral ligaments, and/or endometriotic nodules.
EXAMPLES
[00073] The present disclosure is further explained in the form of following examples. However, it is to be understood that the foregoing examples are merely illustrative and are not to be taken as limitations upon the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Such changes and modifications may be made without departing from the scope of the invention.
Example 1: Lyophilized composition of thymosin alpha-1 for subcutaneous injection (1.6mg/vial)
[00074] Lyophilized composition was prepared according to the components and amounts shown in Table-1.
Batch Size: 7000 vials
Table-1
Ingredient Qty / Vial Batch quantity
Thymosin alpha-1 1.6 mg 11.324 gm
Mannitol 50.0 mg 350.0 gm
Disodium hydrogen phosphate anhydrous 1.059 mg 7.413 gm
Sodium phosphate monobasic dihydrate 0.56 mg 3.920 gm
Water for injection q.s to 1.0 ml 7.0 lit
Excipients were dissolved in about 6.5 litres of water for injection at room temperature to get a clear solution. After the solution was cooled to 8°C or below, thymosin alpha 1 was dissolved in the solution under nitrogen atmosphere and then water was added to adjust the total volume to 7 litres. The bulk solution was sterile filtered to remove particulates, if any. The pH of the bulk solution was 6.8. The resultant solution (1 ml) was filled into vials and lyophilized under standard conditions to yield a white powder. Reconstitution of this lyophilized powder with sterile water for injection (1 ml) provided a liquid composition for use in subcutaneous administration.
Example 2: Treatment of patients with endometriosis with thymosin alpha-1
[00075] Endometriosis is a chronic gynecological condition affecting an estimated 10% of women of reproductive age globally, which amounts to approximately 190 million women worldwide. Endometriosis is characterized by the growth of endometrial-like tissue outside the uterus, leading to debilitating pain, infertility, and diminished quality of life. Despite its prevalence, many women face significant delays in diagnosis and inadequate treatment, underscoring the urgent need for more effective and accessible therapeutic options. Studies have shown that a significant proportion of endometriosis patients report severe pain (VAS score greater than 7). It is estimated that 30-60% of women with endometriosis experience severe pain. About 75% of women with endometriosis experience chronic pelvic pain, and many of them report NRS scores = 7. Severe menstrual pain is one of the hallmark symptoms of endometriosis, with up to 80% of patients reporting significant dysmenorrhea. Many report high NRS scores (= 7) during menstruation. About 30-50% of patients report painful intercourse, and the intensity of this pain often results in NRS scores in the moderate to severe range.
[00076] Study design: A Phase III open-label, multi-centre, randomized, two arm, clinical trial to evaluate efficacy and safety of thymosin alpha-1 + standard of care (SOC) in comparison with SOC alone in patients with confirmed endometriosis was conducted. The study population consists of 120 female patients with confirmed endometriosis. Patients were administered, at each administration, two subcutaneous injections of 1.6 mg thymosin alpha-1, twice weekly for a period of 90 days. Patients were followed-up for 3 months.
Primary endpoints were:
1) To evaluate and compare the efficacy of Thymosin a 1 in combination with SOC versus SOC alone in patients with endometriosis in terms of endometriosis associated pain assessed by a Visual Analogue Scale (VAS) score.
Secondary Endpoints were:
1) To evaluate and compare the efficacy of Thymosin a 1 in combination with SOC versus SOC alone in patients with endometriosis in terms of Dysmenorrhea (DYS) pain scale score, Dyspareunia (DYSP) pain scale score, Non-Menstrual Pelvic Pain (NMPP) pain scale, lower back pain, dyschezia related pain on NRS, and overall size of Endometriosis.
2) To assess and compare the safety of Thymosin a 1 in combination with SOC versus SOC alone.
Results:
i. The mean (SD) VAS Score of the patients at screening visit was 6.35(1.85) which was reduced to 1.88(1.44) by visit 8 showing statistically significant improvement (p-<0.001) for Thymosin Arm compared to SOC arm which was reduced to 2.5(1.04) at visit 8 from 5.88(1.92) at baseline with significant improvement (p-<0.001).
ii. The Mean (SD) NRS for Dysmenorrhea (DYS) pain scale score at the screening visit was 2.22(0.58) which was reduced to 0.44(0.51) by visit 8 for Thymosin arm where as it was 2.04(0.68) at screening visit and 0.78(0.55) at visit 8 for SOC Arm, both the arms demonstrated significant improvement from baseline.
iii. Mean (SD) Dyspareunia at baseline line was 1.7(0.77) and 0.65(0.49) at end of visit8 for Thymosin Arm, whereas 1.33(0.82) at Screening visit and 0.53(0.51) by visit 8 for SOC, with respect to this parameter as well both the arms demonstrated significant improvement from baseline.
iv. Mean (SD) Non-Menstrual Pelvic Pain (NMPP) pain scale at the screening visit was 1.3(0.99) which was reduced to 0.67 (0.59) by visit 8 for Thymosin Arm with a significant P-value of 0.023, demonstrated significant improvement from baseline. Mean value of 1.28(0.89) at Screening visit and 0.94(0.42) by visit 8 for SOC Arm (p-0.135).
v. Mean Dyschezia Pain at baseline is 6.04(2.34) which got reduced to 1.78(1.52) by visit8 for Thymosin Arm and 4.8(2.66) at screening visit and 2.28(1.49) by Visit 8 for the SOC Arm.
vi. Both the arms demonstrated significant improvement from baseline.
Based on the results, the Thymosin alpha-1 arm demonstrated a statistically significant reduction in pain across various parameters like endometrial pain, dysmenorrhea, dyspareunia, non-menstrual pelvic pain, and dyschezia. The improvement in VAS Score is significantly superior in Thymosin Arm compared to SOC Arm. These findings suggest that Thymosin alpha-1 may offer a more effective adjuvant treatment for pain management for endometriosis related pain like dysmenorrhea, dyspareunia, non-menstrual pelvic pain, and dyschezia, compared to the standard of care.
,CLAIMS:1. A pharmaceutical composition for treating endometriosis, comprising:
about 0.1 mg/ml to about 50 mg/ml of thymosin alpha-1;
about 1% to about 8% weight/volume (w/v) stabilizer; and
about 1 mM to about 20 mM buffer.

2. The composition as claimed in claim 1, wherein the composition has a pH between 6.0 and 7.5.

3. The composition as claimed in claim 1, wherein the stabilizer is selected from the group consisting of sucrose, trehalose, (2-hydroxypropyl)-ß-cyclodextrin, mannitol, sorbitol, L-arginine, a pharmaceutically acceptable salt of L-arginine, L-proline, a pharmaceutically acceptable salt of L-proline, glycine, a pharmaceutically acceptable salt of glycine, and a combination thereof.

4. The composition as claimed in claim 1, wherein the buffer is selected from the group consisting of a succinate buffer, a histidine buffer, a sodium acetate buffer, a phosphate buffer, tris (tris(hydroxymethyl) aminomethane), diethanolamine, a citrate buffer, and a combination thereof.

5. The composition as claimed in claim 1, wherein the buffer comprises disodium hydrogen phosphate and monobasic sodium phosphate.

6. The composition as claimed in claim 1, wherein the stabilizer is mannitol and the buffer comprises disodium hydrogen phosphate and monobasic sodium phosphate, the composition comprising:
about 0.5 mg/ml to about 25 mg/ml of thymosin alpha-1;
about 2% to about 7% w/v mannitol;
about 4 mM to about 10 mM disodium hydrogen phosphate buffer; and
about 1 mM to about 8 mM monobasic sodium phosphate buffer.

7. The composition as claimed in claim 6, comprising:
about 1.6 mg/ml of thymosin alpha-1;
about 5% w/v mannitol;
about 7.5 mM disodium hydrogen phosphate buffer; and
about 4.5 mM monobasic sodium phosphate buffer.

8. The composition as claimed in any of claims 1-7 that is a liquid.

9. The composition as claimed in any of claims 1-7 that is a reconstituted solution from a lyophilized composition.

10. A liquid composition that is reconstituted from a lyophilized composition wherein the reconstituted solution comprises:
1.6 mg/ml of thymosin alpha-1;
50 mg/ml of mannitol;
1.059 mg/ml of disodium hydrogen phosphate; and
0.56 mg/ml of monobasic sodium phosphate.

11. A method of treating endometriosis in a subject in need thereof, the method comprising administering an effective amount of thymosin alpha-1 to the subject.

12. The method as claimed in claim 11, wherein the effective amount comprises a dose of thymosin alpha-1 ranging from 0.01 mg/kg/day to 10 mg/kg/day; or
wherein the effective amount comprises a dose of thymosin alpha-1 ranging from 0.5 mg to 10 mg per day; or
wherein thymosin alpha-1 is administered by subcutaneous administration.

13. The method as claimed in claim 11, wherein the endometriosis is endometriosis externa, endometrioma, adenomyosis, adenomyomas, adenomyotic nodules of the uterosacral ligaments, and/or endometriotic nodules.

14. Use of thymosin alpha-1 in the manufacture of a medicament for use in the treatment of endometriosis in a subject.

15. The use as claimed in claimed in 14, wherein the endometriosis is endometriosis externa, endometrioma, adenomyosis, adenomyomas, adenomyotic nodules of the uterosacral ligaments, and/or endometriotic nodules.