Composition for treatment of infertility in a female subject

Abstract

A pharmaceutical composition for treating infertility in a female subject substantially consists of local anesthetic of amide type such as lidocaine, human albumin, viscosity controlling agent selected from recombinant hyaluronic acid and combination of recombinant hyaluronic acid and water-soluble cellulose ether, optionally citrate, glucose, and/or amino acid, water optionally comprising one or more ions selected from the group consisting of sodium, potassium, magnesium, calcium, acetate, chloride, sulfate. Also disclosed is a method of treating infertility by administration of the composition to a female subject in need thereof.

Claims

1. Pharmaceutical composition for treating infertility in a female subject, comprising: 0.1 mg/mL to 2.5 mg/mL local anesthetic of amide type selected from the group consisting of articaine, bupivacaine, cinchocaine, etidocaine, levobupivacaine, lidocaine, mepivacaine, oxetacine, prilocaine, ropivacaine, tolycaine, and trimecaine, 0.1 mg/mL to 20.0 mg/mL human albumin, wherein the human albumin is recombinant human albumin; viscosity controlling agent selected from the group consisting of recombinant hyaluronic acid and combination of recombinant hyaluronic acid and water-soluble cellulose ether; and water optionally comprising one or more ions selected from the group consisting of sodium, potassium, magnesium, calcium, acetate, chloride, and sulfate, wherein the composition is a perturbation solution adapted for perturbation.

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

3. The composition of claim 2, comprising from 0.1 mM/L to 5.0 mM/L of citrate.

4. The composition of claim 2, comprising up to about 5 mg/ml of glucose.

5. The composition of claim 1, wherein the local anesthetic is in form of a pharmaceutically acceptable salt thereof.

6. The composition of claim 5, wherein the pharmaceutically acceptable salt is selected from hydrochloride, hydrobromide, and sulfate.

7. The composition of claim 1, wherein the local anesthetic is lidocaine or lidocaine hydrochloride.

8. The composition of claim 1, comprising from 0.02 mg/ml to 5.0 mg/ml of hyaluronic acid.

9. The composition of claim 1, wherein the cellulose ether is hydroxypropyl-methyl cellulose (HPMC).

10. A method for perturbation of the fallopian tubes of a female subject, comprising providing the pharmaceutical composition of claim 1; and administering a pharmacologically efficient amount of the composition by perturbation of the fallopian tubes of the female subject.

11. The composition of claim 1, wherein the human albumin is of recombinant origin and wherein the composition is free from macromolecules of animal or human origin.

12. The composition of claim 1, wherein the composition comprises: 0.3 to 1.5 mg/mL local anesthetic of amide type selected from the group consisting of articaine, bupivacaine, cinchocaine, etidocaine, levobupivacaine, lidocaine, mepivacaine, oxetacine, prilocaine, ropivacaine, tolycaine, and trimecaine; 0.5 to 10 mg/mL human albumin, wherein the human albumin is recombinant human albumin; viscosity controlling agent selected from the group consisting of recombinant hyaluronic acid and combination of recombinant hyaluronic acid and water-soluble cellulose ether; and water optionally comprising one or more ions selected from the group consisting of sodium, potassium, magnesium, calcium, acetate, chloride, and sulfate.

13. The composition of claim 12, wherein the composition comprises 0.02 mg/ml to 5 mg/ml recombinant hyaluronic acid.

14. A method of treating infertility in a female subject, comprising providing the pharmaceutical composition of claim 1; and administering a pharmacologically efficient amount of the composition to the abdominopelvic cavity of the female subject.

15. The method of claim 14, wherein said female subject has been diagnosed with unexplained infertility.

16. The method of claim 14, wherein said female subject has been diagnosed with endometriosis.

17. The method of claim 14, wherein administration is by perturbation of the fallopian tubes.

18. The method of claim 14, wherein administration is by infusion into the peritoneal cavity by means of a catheter.

19. The method of claim 14, wherein administration is by injection into the peritoneal cavity by means of a syringe.

20. The method of claim 14, wherein administration is by surgical deposition in the abdominal cavity.

21. The method of claim 14, wherein administration is by transdermal absorption via the abdominal wall.

22. Pharmaceutical composition according to claim 1, in dry powderous form for reconstitution with water, wherein said one or more ions selected from the group consisting of sodium, potassium, magnesium, calcium, acetate, chloride, and sulfate form part of the dry powderous form or of said water for reconstitution.

23. The composition of claim 22, wherein the dry powderous composition is in the form of a cryoprecipitate.

24. Sustained-release composition for treating infertility in a female subject, comprising: 0.1 mg/mL to 2.5 mg/mL local anesthetic of amide type selected from the group consisting of articaine, bupivacaine, cinchocaine, etidocaine, levobupivacaine, lidocaine, mepivacaine, oxetacine, prilocaine, ropivacaine, tolycaine, and trimecaine, 0.1 mg/mL to 20.0 mg/mL human albumin, wherein the human albumin is recombinant human albumin; and viscosity controlling agent selected from the group consisting of recombinant hyaluronic acid and combination of recombinant hyaluronic acid and water-soluble cellulose ether, wherein the composition is incorporated into a biodegradable porous polymer microparticle or a microparticle comprising a biodegradable polymer shell.

25. The composition of claim 24, wherein the porous polymer or the polymer shell comprises a biodegradable polymer selected from poly-lactide, poly-glycolide, and poly(lactide-co-glycolide).

26. Pharmaceutical composition for treating infertility in a female subject, consisting essentially of: 0.1 mg/mL to 2.5 mg/mL local anesthetic of amide type selected from the group consisting of articaine, bupivacaine, cinchocaine, etidocaine, levobupivacaine, lidocaine, mepivacaine, oxetacine, prilocaine, ropivacaine, tolycaine, and trimecaine, 0.1 mg/mL to 20.0 mg/mL human albumin, wherein the human albumin is recombinant human albumin; viscosity controlling agent selected from the group consisting of recombinant hyaluronic acid and combination of recombinant hyaluronic acid and water-soluble cellulose ether; and water optionally comprising one or more ions selected from the group consisting of sodium, potassium, magnesium, calcium, acetate, chloride, and sulfate, wherein the composition is a perturbation solution adapted for perturbation.

27. The composition of claim 26, wherein the human albumin is of recombinant origin and wherein the composition is free from macromolecules of animal or human origin.

28. The composition of claim 26, wherein the composition consists essentially of: 0.3 to 1.5 mg/mL local anesthetic of amide type selected from the group consisting of articaine, bupivacaine, cinchocaine, etidocaine, levobupivacaine, lidocaine, mepivacaine, oxetacine, prilocaine, ropivacaine, tolycaine, and trimecaine; 0.5 to 10 mg/mL human albumin, wherein the human albumin is recombinant human albumin; viscosity controlling agent selected from the group consisting of recombinant hyaluronic acid and combination of recombinant hyaluronic acid and water-soluble cellulose ether; and water optionally comprising one or more ions selected from the group consisting of sodium, potassium, magnesium, calcium, acetate, chloride, and sulfate.

29. The composition of claim 28, wherein the recombinant hyaluronic acid is included in an amount of 0.02 mg/ml to 5 mg/ml.

Description

EXAMPLE 1

(1) Aqueous solutions containing 1 mg/ml hyaluronic acid were prepared by mixing 0.1 ml of hyaluronic acid stock solution with 3.9 ml of either 0.5 mg/ml lidocaine or 1.0 mg/ml lidocaine stock solutions (above) in a glass vial. Solutions were mixed by extensive swirling at room temperature. After swirling for 30 min the hyaluronic acid stock solution (gel) had not been dissolved completely in the lidocaine solutions but was still present as a gel.

EXAMPLE 2

(2) The procedure of Example 1 was repeated at 37° C. with a negative result since hyaluronic acid gel was seen to adhere to the glass of the vial. The same experiments performed with plastic vials provided similar results.

EXAMPLE 3

(3) 3.9 ml of the two stock solutions were pipetted into a glass vial and heated to about 40° C. To the solution was added 0.1 ml of 40 mg/ml hyaluronic acid to make a solution of 1 mg/ml of hyaluronic acid. Care was observed not to have the hyaluronic acid sticking to the glass. The vials were vigorously swirled for two min. Inspection of the vials showed that the hyaluronic acid had dissolved completely.

EXAMPLE 4

(4) Stock solutions containing 0.5 mg/ml lidocaine or 1.0 mg/ml lidocaine were prepared and heated to about 40° C. To each of the vials was added the amount of hyaluronic acid listed in Table 1. Care was taken not touch the wall of vials when adding the hyaluronic acid. After swirling for about 5 minutes the gel in all vials had dissolved. An aqueous solution containing 16 mg/ml of lidocaine was like a soft jelly and proved difficult to press through a hypodermic syringe. At a concentration of 8 mg/ml the solution was viscous but easy to handle and press through a hypodermic syringe of same gauge. The osmolarity of the solutions was tested by using a Fiske Model 210 Micro Osmometer.

(5) TABLE-US-00001 TABLE 1 Fluid consistency of hyaluronic acid- containing pertubation solutions Pertubation solutions prepared from stock solutions A and B A. Hyaluronic acid 40 0.2 parts 0.4 parts 0.8 parts 1.6 parts mg/ml B. Lidocaine 0.5 mg/ml 3.8 parts 3.6 parts 3.2 parts 2.4 parts Final concentration of hyaluronic acid (mg/ml) 2 4 8 16 Fluid viscosity Liquid Slightly Viscous Gel-like viscous Osmolarity (mOsm/kg) Not tested 265 246 Not tested

EXAMPLE 5

(6) Hyaluronic acid was exchanged for hydroxypropyl-methyl cellulose (HPMC) as a viscosity controlling agent. It was noted that HPMC is also possible to use to control viscosity but a larger volume is needed. A stock solution of 5% HPMC in WFI was prepared.

(7) TABLE-US-00002 TABLE 2 Fluid consistency of hydroxypropyl methyl cellulose-containing pertubation solutions Pertubation solutions prepared from stock solutions C and D HPMC 0.2 0.8 1.6 2.0 Lidocaine 0.5 mg/ml 3.8 3.2 2.4 2.0 Final concentration of HPMC (mg/ml) 2.5 10 20 25 Fluid viscosity Liquid Liquid Liquid Sligthly viscous Osmolarity (mOsm/kg) Not tested Not tested 329 325

EXAMPLE 6

(8) A pertubation solution (50 ml, Table 3) containing glucose and albumin was prepared as follows.

(9) Under aseptic conditions, NaCl (425 mg), CaCl.sub.2⋅H.sub.2O (16.5 mg), lidocaine (25 mg) and glucose (1.25 g) were dissolved in 35 ml WFI in a glass beaker. The solution was heated to about 40° C. Albumin (1 ml of a 50 mg/ml solution of HSA) was added and the combined solutions swirled until completely mixed. The resulting clear solution was the sterilized by passing it 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 vessels walls. The solution was gently swirled at 40° C. When the hyaluronic stock solution (gel) had fully dissolved in the lidocaine solution the pH was adjusted to 7.0 with sterile 1 M NaOH and the solution diluted with WFI to 50 ml. Osmolarity was measured and determined to be 295 mOsm/kg. The solution was aliquoted to 5 vials each containing 10 ml. The vials were cooled and stored refrigerated at from 4° C. to 8° C. Prior to use the vials were brought to room temperature.

(10) TABLE-US-00003 TABLE 3 Hyaluronic acid-containing pertubation composition (mg/50 ml) (mg/ml per vial) NaCl 425 8.5 KCl 15 0.3 CaCl.sub.2•H.sub.2O 16.5 0.33 Lidocaine 25 0.5 Glucose 1250 25 Albumin 50 1 Hyaluronic acid 400 8

EXAMPLE 7

(11) Comparison of the composition of the invention with a corresponding prior art composition lacking albumin in a murine surgically-induced homologous endometriosis model.

(12) The objective of the study was to assess the efficacy in improving fertility of the composition of the invention and a corresponding prior art composition lacking albumin (“prior art composition”) in an endometrial auto-transplantation model in female C57BL/mice. The composition of the invention of Table 4 was used in the study.

(13) TABLE-US-00004 TABLE 4 Composition of the invention used in the study Component 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 q.s., pH 7.0 Water for injection ad 1.0 ml
Procedure. The study included four groups of n=5 female C57BL mice. Three groups were subjected to unilateral uterine horn resection and auto-transplantation of the endometriotic tissue adjacent to the arterial cascade of the intestinal mesentery while one group was subjected to unilateral uterine horn rejection only and served as sham-control. The female animals' estrous cycle was synchronized prior and following the surgical procedure and the reception potential assessed visually. Fertility of all females was assessed by mating success and subsequent litter size determination performed 6-7 weeks post-surgical procedure. Treatment was carried out by once daily repeated intraperitoneal injections for three successive days of the composition of the invention and the prior art composition to two of the endometrial auto-transplantation groups just prior to mating. The composition was administered at a dose per injection of 20 ml/kg body weight, that is, a dose of 10 mg of lidocaine/kg body weight. The other two groups were injected with a buffer control solution under identical experimental conditions and served as control groups. Animals were clinically observed for a duration of up to 10 weeks. No treatment mortalities were noted.
Results. Females from the group treated with the composition of the invention exhibited higher body weight gain than of the other groups. The difference was even higher when taking into account only the animals in which a copulatory plug was observed and/or suspected as pregnant during the rise in body weight. The results are shown in Table 4.

(14) TABLE-US-00005 TABLE 4 Pregnancy efficacy of the composition of the invention in a C57BL mouse model Operation Pregnant Litter size per Litter per pregnant Litter per (n) Composition animals (no/%) animal/total animal (mean) group (mean) Endometriosis- Invention 3 (60%) 5, 3, 4 (12) 4.0 2.4 induced (5) Endometriosis- Prior art 2 (40%) 3, 2 (5) 2.5 1.0 induced (5) Endometriosis- Placebo 1 (20%) 3 (2) 3.0 0.6 induced (5) Sham-operated Placebo 2 (40%) 5, 6 (11) 5.5 2.2 control (5)
It is evident that the composition of the invention has substantially higher efficacy than the prior art composition, both being superior to placebo. The composition of the invention is even at least equivalent with placebo in the sham-operated control. The composition of the invention thus is capable of restoring fertility to normal levels in the presence of endometriosis.