MXPA97006182A - Use of hyaluronic acid for the treatment of interstic cystitis - Google Patents

Abstract

A method of treatment comprising contacting the internal bladder and associated structures in a mammal having interstitial cystitis, with a solution containing hyaluronic acid having an average molecular weight of not less than 2 x 10-6 Daltons at an effective concentration to treat interstitial cystitis

Description

USE OF HYALURONIC ACID FOR THE TREATMENT OF INTERSTITIAL CYSTITIS. Field of the Invention The present invention relates to a novel method for the treatment of internal bladder and associated structure in a mammal, comprising the step of contacting them with a solution comprising hyaluronic acid having an average molecular weight not less than 2 X 105 Daltons in an effective concentration to treat interstitial cystitis. More particularly, the present invention relates to a novel method for the treatment of internal bladder and associated structures in a mammal having interstitial cystitis, comprising the steps of contacting the internal bladder and associated structures in the mammal that has interstitial cystitis with a solution containing hyaluronic acid having an average molecular weight of 5 X 105 to 3.1 X 106 Daltons, at an effective concentration for the treatment of interstitial cystitis. BACKGROUND OF THE INVENTION In mammals the only tight junctions of epithelial cells on the surface of the bladder are the fundamental mechanism by which the bladder maintains its impermeability. However, the glycosaminoglycan layer on the laminar surface of the bladder wall can be a defense mechanism for the protection of the transitional epithelium from urinary irritants. (Chelsky, M. et al., 1994, Journal of Urology, 151: 346). (This glycosaminoglycan layer consists of mucopolysaccharides attached to a central protein which, in turn, is linked to a central hyaluronic acid line.) This highly hydrophilic and highly viscous glycosaminoglycan layer can protect the transitional epithelium of the bladder from irritants. which are included in the urine, but not limited to pathogens, microcrystals, calcium proteins and carcinogens. (Nickel, JC et al., 1993. Journal of Urology, 149: 716) This glycosaminoglycan layer could also obstruct small molecules not loaded, such as urea, from diffusion through the transitional cell epithelium, so the glycosaminoglycan layer covers the bladder so that it can act as a barrier between the environment within the bladder volume, and the transitional epithelium of the bladder. The bladder can thus protect the transitional epithelium from inflammation, infection, injury, stone formation and carcinogenesis.

Interstitial cystitis is a poorly understood condition of the bladder, for which there is no effective universal treatment program. (Fleishmann, J.D. et al., 1991. Journal of Urology). Symptoms include urgency to urinate, increased frequency in urine, and suprapubic damage, usually revealed by emptying. Other symptoms include arthritis, spasmodic colon and light fever. Individuals with interstitial cystitis may be significantly disabled, and individuals with advanced interstitial cystitis may require major surgery in order to function. Although the etiology of interstitial cystitis remains unexplained, it has been suggested that abnormalities of or deficiencies in the glycosaminoglycan coating layer of the bladder transitional epithelium may be a primary defect (Eldrup J. 1983. British Journal of Urology, 55: 488). These abnormalities or deficiencies may allow for increased permeability of the transitional epithelium (Parsons, El et al, The Journal of Urology, 143: 690) and this increased permeability may allow urinary solutes to gain access to subepithelial tissue and to induce a response Inflammatory and irritant that contributes to the symptoms of interstitial cystitis. There is no standard treatment for interstitial cystitis. Among the treatments used are hydraulic distension of the bladder, oral amitriptyline or sodium pentosanpolysulfate, intravesical instillation of dimethyl sulfoxide, sodium oxychlorosene., silver nitrate, heparin or a composition comprising an angiostatic steroid and pentasanpolisulfate. However, the efficiency of this treatment is variable. Hydraulic distension of the bladder is done under general or spinal anesthesia for one to two minutes at a pressure of 80 to 100 cm H20. In one study, hydraulic distention of the bladder was used for the treatment of interstitial cystitis, less than 55% of treated patients were reported with immediate relief after treatment and only 2% were reported to be relieved six months after treatment ( Hanno PM et al., 1991. Semin. Urology, 9: 143). The instillation of dimethylsulfoxide (DMSO) in the bladder for six to eight weeks results in a 53% proportion in response to DMSO versus 18% response to the placebo, with the highest average response being six months (Pérez-Marrero , R. et al., 1967. Journal of Urology, 98: 671). The pharmacological effects of DMSO include membrane penetration, improved absorption of the drug, anti-inflammatory and analgesic effects, collagen dissolution, muscle relaxation and release of histamine from mast cells. Side effects include increased bladder irritability and breathing with a smell similar to garlic. It has been reported that the results are equivalent to the instillation of DMSO with sodium oxychlorosene (Messing, E.M. et al., 1978. Urology, 12: 381). However, the instillation of oxychlorosene requires anesthesia because of the intense discomfort. Sodium pentosanpolysulphate is a synthetic glycosaminoglycan of low molecular weight, US Pat. No. 4,524,066 to Wolf) and is characterized by having a very low viscosity and high electronegativity. US Patent No. 4,820,693 to Gillespie (Gillespie * 693) discloses a composition and method for arresting angiogenesis and cellular, capillary or membrane filtration, comprising either oral or intravesical administration of an angiostatic steroid and pentosanpolysulfate. The molecular weight of pentosanpolysulfate is between 1.5 X 10a and 5 X 103 Daltons (The Merck Index, Grape edition, 1989, p.7093 in 7090). The molecular weight of the pentosanpolysulfate claimed by Gillespie '693 is between 2 X 103 and 6 X 103 Daltons, and is preferably from about 2 X 103 Daltons. In US Patent No. 4,966,890 to Gillespie (Gillespie J * 890) discloses a composition and method for treating interstitial cystitis comprising either oral or intravesical administration of an angiostatic steroid pentosanpolisulfate. In the '890 Patent, Gillespie describes that pentosanpolysulfate can be used in place of heparin and that he pentosanpolysulfate in combination with an angiostatic steroid cures interstitial cystitis by stopping angiogenesis, cell membrane filtration, capillary leakage or exchange in the bladder. U.S. Patent No. 5,180,715 to Parsons (Parsons' 715) also discloses the use of pentosanpolysulfate for the treatment of interstitial cystitis. In the '715 Patent, Parsons provides data showing that it is more effective for the treatment of interstitial cystitis when the pentosanpolysulfate is administered orally and in doses in excess of 100 mg per day. In the '715 Patent Parsons also suggests, but does not provide data to prove it, that intravesical instillation of pentosanpolisulfate is used for the treatment of interstitial cystitis. In the '715 Parsons patent, it teaches that pentosanpolysulfate can be used in place of heparin and that it acts to block bacterial adhesion to the transitional epithelium of the bladder. , The pentosanpolisulfato is a glycosaminglicano of low viscosity and has been disclosed in Patents 693 of Gillespie,? 890 of Gillespies and on v715 of Parsons. Interstitial cystitis may be related to a defect in the glycosaminoglycan layer of high viscosity on the luminal surface of the bladder, intravesical administration does not provide adequate protection to the transitional epithelium of the bladder and associated structures. Therefore, what is needed is a highly viscous and highly hydrophilic substance which covers the transitional epithelium of the bladder and associated structures. A highly viscous and highly hydrophilic substance should provide a barrier between irritants within the lumen of the bladder and associated structures and the coating of the transitional epithelium and associated structures. The hyaluronic acid (AH) heteropolysaccharide consisting of alternating residues of D-glucuronic acid and N-acetylglucosamine. The AH is a linear polymer with a molecular weight of up to 13 X 106 Daltons. It has been found in the connective tissue, joint synovial fluid, in ocular vitreous humor, in the umbilical cord and is synthesized by some bacteria that include but are not limited to the streptococcal species. The molecular weight of AH inhibits the migration of lymphocytes (Balazs E.A. et al., 1973 In: Biology of Fibroblasts. Academic, Press, pp. 237-252). The phagocytic and chemotactic capacity of neutrophils and leukocytes are also inhibited. (Brandt, K.D. 1974. Clinical Chemical Acta 55: 307). The AH is highly viscous, has a high electronegativity and is highly hydrophilic. The use of HA for the treatment of interstitial cystitis is the present invention provides unexpectedly excellent results in the treatment of interstitial cystitis in a mammal with this condition. SUMMARY OF THE INVENTION Briefly, the present invention comprises a method for the treatment of interstitial cystitis in a mammal with this condition, comprising the steps of contacting the transitional epithelium of the internal surface of the bladder and associated structures, which include the ureters and urethra in a mammal having interstitial cystitis, with a solution containing HA having an average molecular weight of not less than 2 X 105 Daltons at an effective concentration for the treatment of interstitial cystitis. This invention further comprises the addition of various substances to the HA composition including, but not limited to, antibiotics, extracts of bacterial cells, viruses, cytokines and interferons, for use in the treatment of interstitial cystitis, bladder damage, Bladder irritation and bladder infection. It is an object of the present invention to provide a method for the treatment of interstitial cystitis in a mammal with this condition by contacting the transitional epithelium of the internal surface of the bladder and associated structures with a solution containing AH, which has an average molecular weight of not less than 2 X 105 Daltons in an effective concentration for the treatment of interstitial cystitis. It is also another object of the present invention to provide a method for the treatment of traumas, irritation or infection of the lining of the renal pelvis, ureters, bladder or urethra, by contacting the inner surface of the renal pelvis, ureters, bladder and urethra with a solution containing AH, which has an average molecular weight of not less than 2 X 105 Daltons at an effective concentration to treat trauma, irritation or infection of the lining of the renal pelvis, ureters, bladder or urethra. Other objects, features and advantages of the present invention will become apparent on what is read in the following detailed description of the preferred embodiment of the invention when taken in conjunction with the appended claims. Brief Description of the Figures Figure 1 shows the percentage reduction of symptoms in Group A and Group B after 4, 8 and 12 weeks of treatment with hyaluronic acid. Detailed Description of the Invention As used herein, the phrase "inner surface of w refers to the transitional epithelium that delimits the bladder." As used herein, the phrase "associated structures" refers to the ureter and urethra. directed to a method for the treatment of interstitial cystitis in a mammal with interstitial cystitis by contacting the inner surface of the bladder and associated structures with a solution containing AH which has an average molecular weight of not less than 2 × 10 5 Daltons in a concentration Effective for the treatment of interstitial cystitis It has been found that HA and salts thereof having an average molecular weight of not less than 2X10 5 Daltons is unexpectedly useful in the treatment of interstitial cystitis in mammals with interstitial cystitis. this invention has an average molecular weight of not less than 2 × 10 5 Daltons, preferably the AH has a weight or molecular in the range of 5X105 3.1X106 Daltons. More preferably the AH has a molecular weight in the range of 6X105 * 19X106 Daltons. More preferably, the HA has an average molecular weight selected from the group consisting of 6.5X105 Daltons, 8.7X105 Daltons and 1.9X106 Daltons. Several methods are available to obtain fractions of the molecular weight of HA. These include the fractionation of the AH prepared from cartilage, fractionation of AH derived from bacteria that include, without being limited to these, the streptococcal species and the acquisition of fractions of the molecular weight of the AH coming from commercial sources include, without being limited to these, Fluka Chemical Corporation, Ronkonkoma, NY, Genzyme Corporation, Cambridge, MA and Lifecore Biomedical, Inc., Chaska MN. The HA used in the present invention is present in a concentration from about 0.01 mg / ml to about 25 mg / ml. More preferably, AH is present in a concentration from about 0.1 mg / ml to about 2 mg / ml. More preferably the AH is present a concentration from about 0.4 mg / ml to about 1.2 mg / ml. The HA is solubilized in a pharmaceutically acceptable buffer that includes, but is not limited to, physiological saline and buffered phosphate solution. However, it should be understood that any physiological buffer known to those skilled in the art as pharmaceutically acceptable to be in contact with the surface of the bladder and associated structures in a mammal may be employed in the present invention. The HA solution that is employed in the present invention may also include an antibiotic effective for the treatment of interstitial cystitis. The determination of the antibiotic and the amount of the antibiotic to be included in the AH solution within the determination of those skilled in the art. The HA solution that is employed in the present invention may further include an immunotherapeutic agent that includes, but is not limited to, bacterial cell extracts such as mycobacterial cell wall extracts and cell wall extracts of Calmette bacilli. Guerin, viruses, cytokines and interferons. The Ah solution for use in the present invention is infused into the bladder and associated structures in a volume of between about 40 and 60 ml. The amount of AH to be infused into the bladder and associated structures for the present invention is between about 5 mg and 100 mg. More preferably the amount of HA to be infused into the bladder and associated structures is between about 20 mg and 60 mg. More preferably the amount of AH to be infused into the bladder and associated structures is approximately between 35 mg and 45 mg. The HA solution of the present invention can be administered from a container such as a bottle, without being limited thereto. The composition can be administered in the bladder and associated structures using a urinary catheter. However, it will be understood that any method known to those skilled in the art can be employed to contact the inner surface of the bladder and the associated structures of a mammal with a pharmaceutical solution. The HA solution must remain in contact with the bladder and associated structures from approximately e30 minutes to 8 hours. More preferably from 30 minutes to 4 hours. More preferably from 30 minutes to 2 hours. The treatment of interstitial cystitis of a mammal that suffers it with a solution containing AH, by contacting the bladder and the structures associated with AH and the salts thereof having an average molecular weight of not less than 2 X 105 Daltons, provides unexpectedly good results to relieve the symptoms of interstitial cystitis without disturbing the side effects. EXAMPLE 1 ISOLATION, PURIFICATION AND FRACTIONATION OF HYALURONIC ACID The following describes a method for the isolation, purification and fractionation of cartilage hyaluronic acid for use in this invention.

Rooster crest pretreatment The preparation of sodium hyaluronate from frozen or fresh rooster crests involves the following steps: Rooster crests are minced, homogenized, dehydrated in acetone and dried under vacuum. The water content of the acetone discharged is less than 2.0%. The powder is digested enzymatically with papain in a buffered aqueous medium containing cysteine hydrochloride. The resulting mixture is clarified and ultrafiltered using a membrane with a molecular weight exclusion limit of 3 X 10 4 Daltons. The retained clear liquid has a pH between 5.0 and 7.0. The content of mucopolysaccharide is 2.0 and 6.0 mg / ml of sodium hyaluronate, as determined by a glucuronic acid assay. The amino acid content is greater than 6.0 mg / ml, as determined in a ninhydrin assay. Accompanied. Fractionation and Precipitation NaCl (up to 0.1 M) and cetyl-pyridinium chloride (CPC) were added to the clear liquid with stirring. The precipitate is collected by centrifugation and washed three times with 0.01M NaCl with 0.05% CPC. The precipitate is suspended in 0.05M NaCl with 0.05% CPC with shaking and the nebulous supernatant is removed. This procedure is repeated several times using 0.1M NaCl with 0.05% CPC. The precipitate is then dispersed with 0.3 M NaCl with 0.05% CPC with stirring and the extraction is repeated three times. The precipitate is then removed. The clear supernatants are mixed, brought to a concentration of 0.23 M NaCl, CPC is added, the mixture is treated with Celite (R) and filtered. After treatment with Celite (R) the content of sodium hyaluronate is 2.5-5.0 mg / ml as determined by the glucuronic acid test. Isolation of Hyaluronic Acid. The filtrate is ultrafiltered using a membrane with a molecular weight with an exclusion limit of 3 X 104 Daltons and the retained liquid is concentrated. The liquid is precipitated with 95% ethanol and centrifuged. The precipitate is dissolved in 0.1 M NaCl and precipitated again with 95% ethanol. the precipitate is collected and washed obtaining a crude product having an average molecular weight of not less than 2.5 X 105 Daltons. The yield is equivalent to or .6% of the original fresh tissue. Purification of the Hyaluronic Acid Fraction The precipitate is dissolved in pyrogen-free distilled water (10 mg / ml) and ultrafiltered using a membrane with a molecular weight exclusion limit of 2 X 105 Daltons without the addition of supplemental water. This increases the concentration of the molecules having a molecular weight greater than 2 X 105 Daltons. Ultrafiltration is used to reduce the volume to 10% of the original volume. Water is added to the concentrated solution and the operation is repeated twice. The concentrated solution is collected and diluted with water to a concentration of 5mg / ml of hyaluronic acid. NaCl is added until it reaches the solution at 0.1 M concentration and the solution is precipitated with four volumes of 95% ethanol. The precipitate is washed and then dried under vacuum. The purified hyaluronic acid is polydispersed and has an average molecular weight of not less than 2 X 105 Daltons. Methods for further fractionation of this AH into different molecular weight fractions are well known to those skilled in the art. Additional methods for the preparation of purified AH of the molecular weights claimed in this invention are described in US Pat. No. 4,141,973 to Balzas, which is incorporated herein by reference. EXAMPLE 2 PILOT STUDY OF INTERSTITIAL CYSTITIS In this pilot study, five patients with interstitial cystitis received an intravesical instillation of 40 mg of hyaluronic acid having an average molecular weight of 6.5 X 105 Daltons in 40 to 70 ml of sterile saline (USP). ). Recent results for this pilot study are related to the improvement of symptoms based on descent symptoms prior to therapy, damage prior to therapy and urgency prior to therapy. SUBJECT 1 PATIENT JM In patient JM (# 002) with interstitial cystitis, treatment failed with the instillation of intravesical heparin pentosanpolisulphate oral. JM was treated according to the protocol study. Forty mg of AH have an average molecular weight of 6.5 X 105 Daltons (in the range of 5 X 105 to 7.3 X 105 Daltons) in 50 ml of normal saline (USP), administered in the bladder under sterile conditions using a catheter urethral. The catheter is removed and the AH solution is kept in the bladder for 30 minutes. The treatment is repeated weekly for seven weeks. After the 7th treatment, the patient reports a marked improvement in suprapubic damage and an urgency of urine Treatment is repeated four times during the following 17 weeks After the last treatment, patients report a 100% completion In the suprapubic damage and the performance in urgency There is no effect of the AH treatment reported by the patient SUBJECT 2 PATIENT GH In patient GH (# 003) with interstitial cystitis, treatment with propantheline bromide fails (2 -hydroxy-yl) -diisopropyl methylammonium, xanten-9-carboxylate bromide, hydroxychloridene-phenylpropanolamine and guaifenesin The patient GH was treated according to the study protocol Forty mg of AH, which have an average molecular weight of 6.5 X 105 Daltons, in 50 ml of normal saline solution (USP), administered in the bladder under sterile conditions using a urethral catheter, the catheter is removed and the HA solution is maintained in the bladder for 60 minutes. The treatment is repeated 4 times for approximately 12 weeks period. After him. Last treatment, patients report a 100% completion in the suprapubic damage and the realization in urgency. There is no effect of AH treatment reported by the patient.

SUBJECT 3 PATIENT LB In patient GH (# 001) with interstitial cystitis, treatment with intravesical infusion of DMSO and heparin failed. LB was treated according to the study protocol .. Forty mg of AH have an average molecular weight of 6.5 X 105 Daltons, in 50 ml of normal saline (USP), administered in the bladder under sterile conditions using a catheter urethral. The catheter and the solution of AH stays in the bladder for 45 minutes. The treatment is repeated weekly for 5 weeks with significant improvement in symptoms prior to therapy, and damage prior to therapy and urgency prior to therapy. Due to an illness. SUBJECT 4 PATIENT MM Patient MM (# 004) with interstitial cystitis, is treated according to the protocol study. Forty mg of AH have an average molecular weight of 6.5 X 10 Daltons in 50 ml of normal saline (USP), administered in the bladder under sterile conditions using a urethral catheter. The catheter is removed and the HA solution is maintained in the bladder for 50 minutes. The treatment is repeated 9 times over a period of 22 weeks. After the last treatment, the patient reports improvement in symptoms prior to therapy, in damage prior to therapy and in urgency prior to therapy. Although the patient does not report side effects of AH treatment, the patient chooses the discontinuous treatment of AH. SUBJECT 5 PATIENT MS The patient MS (# 006) with interstitial cystitis, is treated according to the protocol study. Forty mg of AH have an average molecular weight of 6.5 X 105 Daltons, in 50 ml of normal saline (USP), administered in the bladder under sterile conditions using a urethral catheter. The catheter is removed and the AH solution is kept in the bladder for 60 minutes. The treatment is repeated weekly for seven weeks. At the end of the 1st week there is marked improvement in symptoms prior to therapy, in damage prior to therapy and in urgency prior to therapy. Four supportive treatments are given during the next 16 months. The marked improvement is maintained throughout the time and at the end of each support treatment. EXAMPLE 3 STUDY OF INTERSTITIAL CYSTITIS Hyaluronic acid (HA) is used for the treatment of interstitial cystitis according to the present invention. Twenty-four humans each had interstitial cystitis, with AH (sodium hyaluronate) having an average molecular weight of 6.5 X 105 Daltons. Inclusion criteria for this study, includes: l. > 18 years old. 2. diagnosis of interstitial cystitis 3. untreated or pretreatment characteristics 4. two or more of the following present findings: a) suprapubic, urethral or perineal injury; b) chronic inflammation or infiltration of mast cells on cystoscopy or biopsy with no evidence of malignancy; c) hydrodistension under anesthesia at a pressure of 80 to 100 cm of H20 with glomerulations (multiple petechiae), bloody effluent and decreased capacity of the bladder; d) sterile urine culture; e) slight decrease on the cystometrogram; f) bladder filling injury (decrease due to emptying). Exclusion criteria for this study, includes: i. benign or malignant bladder tumors 2. evidence of vesicourethral reflux or urethral diverticulum 3. uterine, cervical, vaginal or urethral cancer 4. UTI, vaginitis, prostatitis 5. urethral or bladder stones, active herpes (herpes virus type II) 6. positive urine cytology cystometry capacity > 400 cc, absence of sensory urgency or unstable bladder 8. waking frequency < 5 in 12 hours 9. neurogenic bladder dysfunction 10. patients taking any medication or active treatment for the treatment of interstitial cystitis within 30 days of study registration 11. previous urinary fun 12. pregnant women. Evaluation criteria for this study, including: The symptoms prior to treatment and the effects of AH treatment are assessed using a quality of life symptom registry, a visual analogue pain scale (VAS), an emergency scale of Visual analogue (VAS) and a 72-hour emptying record.

Criteria of results for this study, which include: 1. Complete Response (CR): Improvement of symptoms with a>. 90% decrease in the symptom register prior to treatment, VAS pain registry, VAS urgency scale and the patient does not require medication. 2. Partial Response (PR): Incomplete resolution of symptoms with > 50% decrease in the registry of symptoms prior to treatment, VAS pain registry, VAS urgency scale and the patient continues to require medication for the complete help of the symptoms. 3. Minor response (MR): Incomplete resolution of symptoms with > 50% decrease in the registry of symptoms prior to treatment, VAS pain registry, VAS urgency scale and the patient continues to require medication for the partial help of the symptoms. 4. Deficiency (D): No improvement in symptoms. 5. Withdrawal (R): Patients withdrawn or withdrawn from the study.

Treatment protocols for this study include: Each patient receives an intravesical instillation of 40 mg of HA that has an average molecular weight of 6.5 X 105 Daltons in 50 ml of normal saline (USP). It is introduced under sterile conditions using a urethral catheter in the bladder and sent by bacterial culture. 50 ml of the composition is introduced into the bladder and the catheter is removed. The patient is asked to retain the HA solution for as long as possible. The instillation of AH is given 1 X / week for 4 weeks (induction). If the patient's symptoms are completely resolved within the induction period of 4 weeks, treatment is given monthly until the end of 12 months (care). If the patient's symptoms are partially resolved within the induction period of 4 weeks, the patient is portrayed lX / week for 4 weeks. If after the retreat the patient's symptoms are completely resolved the care treatment is given. If the patient's symptoms are not completely resolved within the induction period of 4 weeks, the treatment is discontinued and the patient is withdrawn from the study. The results obtained are shown in Table I and Table II. Table I shows the results of 14 patients who entered the study several times within the study protocol. Twenty of the 14 patients showed either a complete or partial response to HA treatment after 4 to 28 weeks of treatment. Two of the 14 patients showed no response to AH treatment after 8 weeks of treatment. TABLE I TREATMENT OF HYALURONIC ACID (AH) AGAINST INTERSTITIAL CYSTITIS Table II shows the results of 24 patients who completed 24 weeks, 23 patients who completed 32 weeks, 20 patients who completed 40 weeks and 16 patients who completed 48 patients of the AH treatment protocol. At the end of the induction period (week 4), 4 patients showed a complete response, 10 a partial response, 9 a minor response and 1 no response to AH treatment. One patient did not complete the induction period. Of the 18 patients remaining in the study after 24 weeks of treatment with AH, 6 showed a complete response, 6 a partial response, 4 a minor response and 2 did not respond. Of the 9 patients remaining in the study, after 48 weeks of treatment with AH, 2 showed a complete response, 6 a partial response and 1 with a smaller response. Of the 7 patients who withdrew from the study, 3 withdrew from treatment due to poor treatment and 4 withdrew due to protocol violations.

TABLE II RESPONSE RATE TO THE TREATMENT OF HYALURONIC ACID (AH) AGAINST INTERSTITIAL CYSTITIS The data in Table I and Table II show that contacting the internal bladder and associated structures with a solution containing AH, which has an average molecular weight of 6.5 X 105 Daltons, being unexpectedly effective in relieving the symptoms of interstitial cystitis. This is the method of treatment of AH against interstitial cystitis of the present invention, which results in significant and unexpected improvement in symptoms prior to therapy, damage prior to therapy and urgency prior to therapy. EXAMPLE 4 A patient with interstitial cystitis is treated according to the following protocol. Forty mg of AH have an average molecular weight of 8.7 X 105 Daltons, in 50 ml of normal saline (USP), administered in the bladder under sterile conditions using a urethral catheter. The catheter is removed and the AH solution is kept in the bladder for 60 minutes. The treatment is repeated weekly and there is a marked improvement in symptoms prior to therapy, in damage prior to therapy and in urgency prior to therapy. EXAMPLE 5 A patient with interstitial cystitis is treated according to the following protocol. Forty mg of AH have an average molecular weight of 1.9 X 105 Daltons, in 50 ml of normal saline (USP), administered in the bladder under sterile conditions using a urethral catheter. The catheter is removed and the AH solution is kept in the bladder for 60 minutes. The treatment is repeated weekly and there is a marked improvement in symptoms prior to therapy, in damage prior to therapy and in urgency prior to therapy. EXAMPLE 6 STUDY OF INTERSTITIAL CYSTITIS Hyaluronic acid is used for the treatment of interstitial cystitis in humans having said condition, according to the present invention. Each patient in this study knows the criteria established by the National Institute of Arthritis, Diabetes, Digestive and Kidney Diseases (NIDDK), for having interstitial cystitis. Inclusion criteria for this study, includes: 1. >; 18 years old. 2. diagnosis of interstitial cystitis 3. untreated or pretreatment characteristics 4. record of > 6 on a Symptom Assessment Record prior to therapy 5. record of > 4 on a VAS damage record prior to therapy 6. record of > 4 about a VAS Urgency Record prior to therapy Exclusion criteria for this study, includes: 1. bladder capacity of > 350 ce on a cystometry taken 2. duration of symptoms < 9 months 3. absence of nocturia 4. relief symptoms by antimicrobials, urinary antiseptics, anticholinergic or antispasmodic 5. waking frequency < 8 in 24 hours. 6. cystitis or bacterial prostatitis within a period of three months 7. low urethral calculi or bladder, active genital herpes 8. uterine, cervical, vaginal or urethral cancer 9. radiation, tuberculous and chemical cystitis, - 10. bladder tumors benign or malignant 11. neurogenic bladder dysfunction. 12. patients taking any medication or active treatment for the treatment of interstitial cystitis within 30 days of study registration 13. previous urinary fun 14. pregnant women. Evaluation criteria for this study, including: Pre-treatment symptoms and the effects of AH treatment are assessed using a quality of life symptom questionnaire, a visual analogue pain scale (VAS), an emergency scale of Visual analogue (VAS) and a 72-hour emptying record.

Criteria of results for this study, which include: 1. Complete Response (CR): Improvement of symptoms with a 75% decrease in the registry of symptoms prior to treatment, VAS pain registry, VAS urgency scale and the patient does not require medication. 2. Partial Response (PR): Incomplete resolution of symptoms and 50-74.99% decrease in the registry of symptoms evaluation prior to therapy, decreases the registry of symptoms prior to treatment, VAS pain registry prior to treatment and scale of emergency VAS prior to treatment. 3. Minor Response (MR): Incomplete resolution of symptoms with a record of symptom evaluation prior to therapy, VAS pain record, and VAS urgency scale. 4. Deficiency (D): No improvement in symptoms. 5. Withdrawal (R): Patients withdrawn or withdrawn from the study. Treatment protocols for this study, includes: Eleven humans, each have interstitial cystitis, are sequentially assigned to receive intravesical instillation of AH, which has an average molecular weight of 8.7 X 105 Daltons (Group A) or 1.9 X 106 Daltons (Group B). Patients in group A and group B each receive an intravesical instillation of 40 mg of AH from group A of hyaluronic acid or hyaluronic acid from group A in 50 ml of normal saline (USP). It is introduced under sterile conditions using a urethral catheter in the bladder and sent by bacterial culture. 50 ml of the composition is introduced into the bladder and the catheter is removed. The patient is asked to retain the HA solution for as long as possible. The instillation of AH is given 1 X / weeks for 4 weeks (induction), followed by 1X / 4 weeks for 8 weeks (prevention). The therapy is discontinued at 12 weeks after the first instillation of AH. The efficiency of HA in the treatment of interstitial cystitis is assessed at 4.8 and 12 weeks after the first HA instillation using a symptom evaluation record, a VAS injury record; and a VAS emergency registry. The results for the 11 patients entered the study several times within the study period, Table III, Table IV, Table V and Figure 1 are shown. Table III shows the response of each of the patients in Group A and in Group B to AH treatment after 4.8 and 12 weeks. The assessment of the response is based on the calculation of the average of the three registers: the symptom evaluation record, the VAS damage scale and the urgency scale. These data show that HA has an average molecular weight of 8.6 X 105 Daltons (Group A) and 1.9 X 106 Daltons, they are effective in revealing the symptoms of interstitial cystitis in a majority of treated patients. TABLE III TREATMENT OF HYALURONIC ACID (AH) AGAINST INTERSTITIAL CYSTITIS Table IV shows the absolute values obtained in the symptom evaluation register (SS), the VAS damage scale (P), and the VAS urgency scale (U) before treatment (week 0), at the end of the period of induction (week 4) and after weeks 8 and 12 of the preventive treatment. At the end of the induction period of week 4, the complete data is available for 11 patients under study. Of these 11, 4 in group A and 5 in group B, they show an improvement in the average% (SS + P + U). One patient in group A and 1 in group B showed increased symptoms after 4 weeks of treatment with AH. Of the 7 patients who finished the prevention period of 8 weeks (week 12), each showed an improvement in the average% (SS + P + U). of these 7, 3 are in group A and 4 are in Group B. Table IV) 3 ¡3 * d h i X s 8 3 8 ! Table V shows a mean standard deviation ± of the percentage change of symptoms prior to therapy in the symptom record, VAS damage scale and VAS urgency scale for Group A and for Group B, then in patients with 4, 8 12 weeks of treatment. Patients in both treatment groups, Group A and Group B, show a decrease in symptoms prior to therapy when they are treated according to the HA method of the present invention. TABLE V PERCENTAGE CHANGE OF THE BASE LINE IN THE PATIENT EVALUATION BY THE TREATMENT GROUP (-) values that indicate a decrease in symptoms.

Figure 1 graphically shows the percentage reduction in symptoms prior to therapy in patients in Group A and in patients in Group B after 4, 8 and 12 weeks of treatment with AH.

The data in Tables IV and in Figure I demonstrate that, in patients having interstitial cystitis, contacting the bladder and structures associated with AH according to the present invention results in a significant and unexpected improvement in symptoms prior to therapy, damage prior to therapy and in urgency prior to therapy. These data also show that each of the AH solutions uses 6.5 X 105 Daltons, 8.7 X 105 Daltons and 1.9 X 105 Daltons, which are unexpectedly effective amounts in the treatment of interstitial cystitis. Although the invention has been described with reference to particular means, materials and examples, it should be understood that the invention is not limited to particular disclosures and that it extends to anything equivalent within the scope of the claims.

Claims (7)

1. Novelty of the Invention Having described the present invention is considered as a novelty and therefore the content of the following claims is claimed as property 1. A method for the treatment of interstitial cystitis, comprising the step of contacting the epithelium of the internal part of the bladder and associated structures in a mammal having interstitial cystitis, with a solution containing hyaluronic acid having an average molecular weight of not less than 2 X 105 Daltons at an effective concentration to treat interstitial cystitis.
2. 2. The method according to claim 1, characterized in that the hyaluronic acid has a molecular weight in the range of 5 X 105 Daltons to 3.1 X 106 Daltons.
3. 3. The method according to claim 2, characterized in that the hyaluronic acid has a molecular weight in the range of 6 X 10 Daltons to 1.2 X 106 Daltons.
4. 4. The method according to claim 3, characterized in that the hyaluronic acid has an average molecular weight of 6.5 X 105 Daltons.
5. 5. The method according to claim 3, characterized in that the hyaluronic acid has an average molecular weight of 8.7 X 105 Daltons.
6. 6. The method according to claim 2, characterized in that the hyaluronic acid has a molecular weight in the range of 1.2 X 106 Daltons to 3.1 X 106 Daltons. The method according to claim 3, characterized in that the hyaluronic acid has an average molecular weight of 1.9 X 106 Daltons.