Compositions for treating infertility in female subjects

Abstract

A pharmaceutical composition for the treatment of infertility in a female subject consisting essentially of an amide-based local anesthetic such as lidocaine, human albumin, a viscosity control agent selected from the group consisting of recombinant hyaluronic acid and a combination of recombinant hyaluronic acid and a water-soluble cellulose ether, optionally citrate, glucose, and / or an amino acid, water, and optionally one or more ions selected from sodium, potassium, magnesium, calcium, acetate, chloride ion and sulfate. Also disclosed are methods of treating infertility by administering the compositions to a female subject in need thereof.

Description

Technical Field

[0001] The present invention relates to a pharmaceutical composition for treating infertility, such as endometriosis-related infertility and unexplained infertility, and also relates to a method of treatment using the composition and a method for preparing the composition. Background Art

[0002] There are many biological reasons for infertility. For example, infertility may be caused by abnormal female hormone levels or ovulatory dysfunction, or by male sperm production dysfunction. However, a large number of couples are diagnosed with unexplained infertility. Artificial insemination is usually attempted as the first treatment for couples with unexplained infertility, and many couples are still waiting for in vitro fertilization (IVF). Tubal perturbation, i.e., Fallopian tube perturbation, has been shown to increase the chances of successful pregnancy in couples with unexplained infertility and patients diagnosed with early endometriosis (Johnson et al., (2005), Cochrane Database Syst Rev, 18(2):CD003718).

[0003] Endometriosis is a gynecological disease characterized by the presence outside the uterine cavity of tissue that is histologically identical to the endometrium, the membrane that lines the uterus of mammals. Endometriosis is associated with infertility and may be an underlying cause in many women with unexplained infertility.

[0004] Lidocaine is a well-known local anesthetic with the following chemical formula:

[0005]

[0006] It is known to have membrane stabilizing, antiarrhythmic and anti-inflammatory properties. In the present application, the term "lidocaine" includes its pharmaceutically acceptable salts, such as the hydrochloride, which is preferably administered in this form. Lidocaine has membrane stabilizing, antiarrhythmic and anti-inflammatory properties. Intratubal perfusion with low doses of lidocaine before artificial authorization has been shown to increase the pregnancy rate of couples with unexplained infertility (Edelstam, G. et al., (2008), Human Reproduction, Vol. 23, No. 4, pp. 852-856).

[0007] US2002 / 00421132A1 discloses an aqueous supplemented culture medium for culturing mammalian gametes and embryonic tissues comprising one or more of human albumin, fermented hyaluronic acid and citrate.

[0008] Hyaluronic acid, also known as acetylated hyaluronan, is a naturally occurring polymer of repeating disaccharide units of N-acetylglucosamine and D-glucuronic acid. It is widely distributed throughout the body and aids in cell proliferation and migration. It is also a component of the extracellular capsule of Group A Streptococci. Hyaluronic acid has found use in a variety of medical applications.

[0009] There is a need for pharmaceutical compositions and / or methods for effectively treating infertility, such as endometriosis-related infertility.

[0010] Purpose of the Invention

[0011] An object of the present invention is to provide a pharmaceutical composition which enhances fertility and can be used to treat infertility, such as unexplained infertility or infertility in patients diagnosed with endometriosis.

[0012] Another object of the present invention is to provide a method for treating unexplained infertility and a method for treating infertility in a patient diagnosed with endometriosis by administering a composition of the above kind to a person in need thereof.

[0013] Other objects of the present invention will become apparent from the following summary of the invention, several examples describing preferred embodiments thereof, and the appended claims. Summary of the invention

[0014] Various aspects of the invention are fully disclosed in the appended claims.

[0015] According to the present invention, a composition for treating infertility (e.g., infertility in patients diagnosed with endometriosis) and for treating unexplained female infertility is disclosed. The composition of the present invention can be injected or implanted into the abdominal pelvic cavity, particularly by injection through the fallopian tube through the cervix using a subcutaneous syringe, also known as tubal injection.

[0016] In one aspect, the present invention provides a pharmaceutical composition comprising an amide local anesthetic selected from articaine, bupivacaine, cinchocaine, etidocaine, levobupivacaine, lidocaine, mepivacaine, oxethicaine, prilocaine, ropivacaine, tococaine and trimecaine or a pharmaceutically acceptable salt thereof, human albumin and hyaluronic acid, and optionally citrate, glucose and / or amino acids, in an aqueous medium, in particular water, or in the form of a powder reconstituted with an aqueous medium, in particular water. Human albumin or hyaluronic acid or both are preferably of recombinant origin. Pharmaceutically acceptable salts include, but are not limited to, hydrochlorides, hydrobromides and sulfates. Lidocaine, in particular in the form of its hydrochloride, is a preferred local anesthetic of the present invention. Another preferred local anesthetic of the present invention is prilocaine.

[0017] The pharmaceutical composition of the present invention is more effective than previously known compositions comprising lidocaine or lidocaine hydrochloride and an aqueous solution of an inorganic salt and / or an organic salt for controlling osmotic pressure, such as Ringer's solution containing sodium chloride. The pharmaceutical composition of the present invention provides a higher fertility rate than the known lidocaine compositions of the above type.

[0018] In the pharmaceutical composition of the present invention, the preferred concentration of the local anesthetic of the present invention, such as lidocaine or lidocaine hydrochloride, is 0.1-2.5 mg / ml, more preferably 0.3-1.5 mg / ml, and most preferably about 1.0 mg / ml.

[0019] To prepare the composition of the invention, the local anesthetic of the invention, in particular lidocaine or lidocaine hydrochloride, is mixed with human albumin and hyaluronic acid and optionally citrate, glucose and / or amino acids in any order, either dry or wet.

[0020] The human albumin of the present invention is preferably of recombinant origin. In the pharmaceutical composition of the present invention, recombinant human albumin provides many advantages over human serum albumin (HAS) of natural origin. By using recombinant human albumin instead of HAS of plasma / blood origin, the potential risk of blood-borne components or infectious particles contaminating the pharmaceutical composition is eliminated. Moreover, recombinant proteins vary less than proteins isolated from biological sources such as blood. Pharmaceutical compositions comprising recombinant human albumin are therefore more easily standardized. The production of recombinant human albumin is well known in the art. In one embodiment, recombinant human albumin is derived from genetically modified yeast, which is modified to express human albumin.

[0021] The preferred concentration of human albumin in the pharmaceutical composition of the present invention is 0.1-20.0 mg / ml, more preferably 0.5-10.0 mg / ml.

[0022] According to a preferred aspect of the present invention, the pharmaceutical composition comprises a viscosity control agent. The viscosity control agent of the present invention is an agent that lacks a pharmacological effect on the condition to be treated. In addition, the viscosity control agent of the present invention is preferably biodegradable.

[0023] The preferred viscosity control agent of the present invention is hyaluronic acid, which term includes its pharmaceutically acceptable salts such as sodium salt, potassium salt, magnesium salt or calcium salt. Most preferred is the sodium salt of hyaluronic acid. Hyaluronic acid is a high molecular weight mucopolysaccharide.

[0024] The hyaluronic acid of the present invention is preferably of recombinant origin. Like recombinant human albumin, the use of recombinant hyaluronic acid can achieve standardization and provide higher safety and stability of the pharmaceutical composition.

[0025] US2002 / 0042132 A1 discloses that citrate does enhance the properties of recombinant human albumin. For the present invention, any citrate known in the art for use in pharmaceutical compositions and / or culture media can be used. Examples of useful citrates include sodium citrate, calcium citrate, citric acid and combinations thereof. Sodium citrate is preferably used. The preferred concentration of the optional citrate in the pharmaceutical composition of the present invention is 0.1-5.0 mM / L, more preferably 0.1-1.0 mM / L.

[0026] According to another aspect of the present invention, the composition comprises glucose. In the composition, glucose controls osmotic pressure and constitutes an energy source. The glucose content is preferably up to about 5% by weight. If it is desired that the composition is isotonic with serum, then the contribution of other components to osmotic pressure is taken into account accordingly, and the content of glucose is selected.

[0027] According to the methods of the present invention, the composition is administered topically to the tissue in need thereof rather than systemically.

[0028] To prepare the composition of the present invention, an amide local anesthetic, particularly lidocaine or a pharmaceutically acceptable salt thereof, and other ingredients are dissolved in water.

[0029] In one embodiment, the pharmaceutical compositions of the invention are free of non-recombinant macromolecules or macromolecules of animal or human origin.

[0030] Another embodiment of the pharmaceutical composition of the present invention comprises a viscosity control agent in addition to hyaluronic acid for regulating the viscosity of the composition. Compared with the pharmaceutical composition with lower viscosity, the pharmaceutical composition with higher viscosity has the advantage of extending the exposure of the pharmaceutical composition to the treated tissue. Any pharmaceutically acceptable viscosity modifier known in the art can be used. In a preferred embodiment, the viscosity modifier is a cellulose ether, particularly hydroxypropyl methylcellulose (HPMC).

[0031] The viscosity of the pharmaceutical composition is preferably adjusted to 40 cp (centipoise) to 600 cp, more preferably 100-400 cp. When hyaluronic acid is used as the only viscosity modifier in the pharmaceutical composition of the present invention, the preferred concentration of hyaluronic acid is 0.02 mg / ml to 2 mg / ml or 3 mg / ml or 5 mg / ml, more preferably 0.1 mg / ml to 1 mg / ml or 0.2 mg / ml to 0.8 mg / ml. When hyaluronic acid is used as a viscosity control agent together with a cellulose ether, particularly HPMC, its concentration and the concentration of the cellulose ether are suitable for providing the composition with the above-mentioned preferred viscosity.

[0032] In another aspect, the present invention provides a method for treating human infertility, comprising providing a pharmaceutical composition according to the present invention and administering a certain amount thereof to the human, particularly to the abdominal cavity or intraperitoneum. Preferably, intraperitoneal administration is performed via the fallopian tube. Administration via the fallopian tube may be performed, for example, by fallopian tube perturbation, see Edelstam, G. et al., (2008), Human Reproduction, Vol. 23, No. 4, pp. 852-856; K. et al., (2012), Human Reproduction, Vol. 27, No. 3, pp. 695-701). The amount of the composition administered is from about 1 ml to about 40 ml, more preferably from 2 ml to 25 ml. For example, a person in need of treatment may have been diagnosed with unexplained infertility or endometriosis.

[0033] Intraperitoneal administration can be, for example, by injecting the composition of the present invention into the peritoneal cavity through a syringe, by infusion into the peritoneal cavity through a catheter, by surgical deposition in the peritoneal cavity, by transdermal absorption through the abdominal wall or by subcutaneous deposition. Sustained or delayed release formulations known in the art comprising the pharmaceutical composition of the present invention can be surgically implanted in the peritoneal cavity or injected therein, for example in the form of microparticles comprising the composition or comprising the composition except its water component, or they can be deposited subcutaneously in the abdomen. Transdermal absorption can be achieved by, for example, a transdermal cotton pad covered with the composition of the present invention. The viscosity of the composition of the present invention used with the pad can be exceptionally significantly higher than 600cp, for example 1000 or 2000cp or even higher.

[0034] The composition is preferably administered to the peritoneal cavity in a volume of 5-10 ml to 20-40 ml, especially about 10 or 20 ml.

[0035] The invention will now be described in more detail by reference to a number of preferred embodiments. DETAILED DESCRIPTION

[0036] Material

[0037] A stock solution of 0.5 mg / ml lidocaine in Ringer's acetate was prepared as follows: NaCl (8.5 g), KCl (0.3 g), CaCl 2 ·H 2 O (0.33 g) and lidocaine (0.5 g) were dissolved in 800 ml of water for injection (WFI). The pH was adjusted to 7.0 with NaOH and water was added to 1000 ml.

[0038] A stock solution of 1.0 mg / ml lidocaine in Ringer's acetate was prepared as follows: NaCl (8.6 g), KCl (0.3 g), CaCl 2 ·H 2O (0.33 g) and lidocaine (1.0 g) were dissolved in 800 ml WFI. The pH was adjusted to 7.0 with NaOH and water was added to 1,000 ml. A hyaluronic acid stock solution (gel) was prepared containing 40 mg / ml hyaluronic acid in WFI.

[0039] Example 1

[0040] An aqueous solution containing 1 mg / ml hyaluronic acid was prepared by mixing 0.1 ml of hyaluronic acid stock solution with 3.9 ml of 0.5 mg / ml lidocaine or 1.0 mg / ml lidocaine stock solution (described above) in a glass vial. The solution was mixed by extensive vortexing at room temperature. After vortexing for 30 minutes, the hyaluronic acid stock solution (gel) was not completely dissolved in the lidocaine solution, but was still present in gel form.

[0041] Example 2

[0042] The procedure of Example 1 was repeated at 37°C and the result was negative as the hyaluronic acid gel was seen adhering to the glass of the vial. The same experiment was performed using plastic vials and similar results were obtained.

[0043] Example 3

[0044] Pipette 3.9 ml of both stock solutions into a glass vial and heat to approximately 40°C. Add 0.1 ml of 40 mg / ml hyaluronic acid to the solution to make a 1 mg / ml hyaluronic acid solution. Be careful not to let the hyaluronic acid stick to the glass. Vortex the vial vigorously for 2 minutes. Check the vial to show that the hyaluronic acid is completely dissolved.

[0045] Example 4

[0046] A stock solution containing either 0.5 mg / ml lidocaine or 1.0 mg / ml lidocaine was prepared and heated to about 40°C. To each vial was added the amount of hyaluronic acid listed in Table 1. Be careful not to touch the walls of the vial when adding the hyaluronic acid. After vortexing for about 5 minutes, the gel in all vials had dissolved. The aqueous solution containing 16 mg / ml lidocaine was like a soft gel and proved difficult to inject through a hypodermic syringe. At a concentration of 8 mg / ml, the solution was viscous but easy to handle and was injected through a hypodermic syringe of the same size. The osmotic pressure of the solution was tested using a Fiske 210 microosmometer.

[0047] Table 1. Fluid consistencies of tubal perfusion solutions containing hyaluronic acid.

[0048]

[0049] Example 5

[0050] Replace hyaluronic acid with hydroxypropyl-methylcellulose (HPMC) as the viscosity control agent. As mentioned above, HPMC can also be used to control viscosity, but requires a larger volume. Prepare a stock solution of 5% HPMC in WFI.

[0051] Table 2. Fluid consistencies of tubal instillation solutions containing hydroxypropyl methylcellulose.

[0052]

[0053] Example 6

[0054] A solution for tubal perfusion containing glucose and albumin (50 ml, Table 3) was prepared as follows.

[0055] Under sterile conditions, NaCl (425 mg), CaCl 2 ·H 2 O (16.5 mg), lidocaine (25 mg) and glucose (1.25 g) were dissolved in 35 ml of WFI. The solution was heated to about 40°C. Albumin (1 ml of 50 mg / ml HAS solution) was added and the combined solution was vortexed until completely mixed. The resulting clear solution was sterilized by passing through a 0.4 micron filter. Then 10 ml of sterile hyaluronic acid stock solution (gel) was aseptically added while preventing it from adhering to the wall. The solution was gently vortexed at 40°C. When the hyaluronic acid stock solution (gel) was completely dissolved in the lidocaine solution, the pH was adjusted to 7.0 with sterile 1 M NaOH and the solution was diluted to 50 ml with WFI. The osmotic pressure was measured and determined to be 295 mOsm / kg. The solution was aliquoted into 5 bottles, each containing 10 ml. The vials were cooled and refrigerated at 4°C to 8°C. The vials were placed at room temperature before use.

[0056] Table 3. Hyaluronic acid-containing fallopian tube injection composition

[0057]

[0058] Example 7

[0059] The compositions of the invention were compared with corresponding prior art compositions lacking albumin in a murine surgically induced syngeneic endometriosis model.

[0060] The aim of this study was to evaluate the efficacy of the compositions of the invention and corresponding albumin-deficient prior art compositions ("prior art compositions") in improving fertility in an endometrial autograft model in female C57BL / mice. The compositions of the invention of Table 4 were used in the study.

[0061] Table 4. Compositions of the invention used in the study

[0062] Components Amount (mg / ml) Sodium chloride 7.8 Potassium chloride 0.3 Calcium chloride dihydrate 0.33 Lidocaine Hydrochloride 0.5 albumin 1.0 Hyaluronic acid 1.0 Sodium hydroxide Appropriate amount, pH 7.0 Water for injection Add to (ad) 1.0ml

[0063] Procedure: The study included four groups of n=5 female C57BL mice. Three groups received unilateral uterine horn resection and autologous transplantation of endometriotic tissue adjacent to the arterial cascade of the mesentery, while one group received only unilateral uterine horn resection and served as a sham-control. The estrous cycle of the female animals was synchronized before and after the surgical procedure, and the receptivity was visually assessed. The fertility of all females was evaluated by mating success and subsequent litter size determination 6 to 7 weeks after surgery. Treatment was performed by repeated intraperitoneal injections of the composition of the present invention and the prior art composition once a day for three consecutive days before mating to two endometrial autologous transplant groups. The composition was administered at a dose of 20 ml / kg body weight per injection, i.e., a dose of 10 mg lidocaine / kg body weight. The other two groups were injected with a buffered control solution under the same experimental conditions and served as a control group. Clinical observation of the animals lasted for up to 10 weeks. No treatment deaths were found.

[0064] Results. Females from the group treated with the composition of the invention showed a higher weight gain than the other groups. The difference was even higher when only animals observed to be mated and / or suspected of being pregnant during the weight gain period were considered. The results are shown in Table 4.

[0065] Table 4. Pregnancy efficiency of the composition of the present invention in C57BL mouse model

[0066]

[0067] It is clear that both the composition of the invention and the composition of the prior art are superior to the placebo, and the efficacy of the composition of the invention is significantly higher than that of the composition of the prior art. In the sham control, the composition of the invention is even at least comparable to the placebo. Therefore, the composition of the invention is able to restore fertility to normal levels in the case of endometriosis.

Claims

1. A pharmaceutical composition for treating infertility in a female subject by administering the composition to a female subject, comprising: - a local anesthetic of the amide type selected from the group consisting of articaine, bupivacaine, cinchocaine, etidocaine, levobupivacaine, lidocaine, mepivacaine, oxethicaine, prilocaine, ropivacaine, tococaine and trimecaine, - human albumin; - a viscosity control agent selected from the group consisting of recombinant hyaluronic acid and a combination of recombinant hyaluronic acid and a water-soluble cellulose ether; - Water, optionally comprising one or more ions selected from the group consisting of sodium, potassium, magnesium, calcium, acetate, chloride, sulfate.

2. The composition of claim 1, additionally comprising citrate, glucose and / or amino acids.

3. The composition according to claim 1 or 2, wherein The human albumin is of recombinant origin.

4. The composition according to any one of claims 1 to 3, wherein The local anesthetic is in the form of a pharmaceutically acceptable salt thereof.

5. The composition according to claim 4, wherein The pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, hydrobromide, and sulfate.

6. The composition according to any one of claims 1 to 5, wherein The local anesthetic is lidocaine or lidocaine hydrochloride.

7. The composition of any one of claims 1-6, comprising 0.1-2.5 mg / ml of local anesthetic, about 1.0 mg / ml of local anesthetic.

8. The composition of any one of claims 1-7, comprising 0.02-5.0 mg / ml hyaluronic acid.

9. The composition of any one of claims 2-8, comprising 0.1-5.0 mM / L citrate.

10. The composition of any one of claims 2-9, comprising up to about 5 mg / ml glucose.

11. The composition according to any one of claims 1 to 10, wherein The cellulose ether is hydroxypropyl-methylcellulose (HPMC).

12. Use of the composition according to any one of claims 1 to 11 in the preparation of a medicament for perfusion of the fallopian tubes of female subjects.

13. Use of the composition according to any one of claims 1 to 11 in the preparation of a medicament for treating infertility in female subjects.

14. Use of the composition according to any one of claims 1 to 11 in the preparation of a medicament for treating infertility in a female subject, comprising administering an abdominopelvic pharmacologically effective amount of the composition to the female subject.

15. The use according to claim 14, wherein: The female subject has been diagnosed with unexplained infertility.

16. The use according to claim 14 or 15, wherein: The female subject has been diagnosed with endometriosis.

17. The use according to any one of claims 14 to 16, wherein: Drug administration is achieved through tubal injection.

18. The use according to any one of claims 14 to 16, wherein: Administration is accomplished by infusion into the peritoneal cavity via a catheter.

19. The use according to any one of claims 14 to 16, wherein: Administration is accomplished by injection into the peritoneal cavity via syringe.

20. The use according to any one of claims 14 to 16, wherein: Administration is achieved by surgical deposition into the peritoneal cavity.

21. The use according to any one of claims 14 to 16, wherein: Administration is achieved by transdermal absorption through the abdominal wall.

22. A pharmaceutical composition in dry powder form, in particular in cryoprecipitated form, for reconstitution with water, for treating infertility in a female subject by administering an aqueous composition to the female subject, the composition comprising: - a local anesthetic of the amide type selected from the group consisting of articaine, bupivacaine, cinchocaine, etidocaine, levobupivacaine, lidocaine, mepivacaine, oxethicaine, prilocaine, ropivacaine, tococaine and trimecaine, - human albumin; - a viscosity control agent selected from the group consisting of recombinant hyaluronic acid and a combination of recombinant hyaluronic acid and a water-soluble cellulose ether; - One or more ions selected from the group consisting of sodium, potassium, magnesium, calcium, acetate, chloride, sulfate.

23. The pharmaceutical composition of claim 22 for sustained release, said composition being incorporated into microparticles of a biodegradable porous polymer or microparticles containing a biodegradable polymer shell.

24. The pharmaceutical composition of claim 23, wherein The porous polymer or polymer shell comprises a biodegradable polymer selected from the group consisting of poly-lactide, poly-glycolide, poly[lactide-co-glycolide].

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