# Patent

## Title
Method of treating neurogenic overactive bladder in a mammal or method of treating non-psychological stress-related bladder dysfunction in a female mammal by administering at least on jasmonate

## Abstract

A formulation or composition contractility comprising jasmonate for modulating bladder and/or treating bladder dysfunction, particularly an overactive bladder in a mammal, particularly a human and use of jasmonate for treating bladder dysfunction is provided.

## Background
TECHNICAL FIELD
A formulation or composition comprising jasmonate for treating bladder dysfunction, particularly an overactive bladder in a mammal, particularly a human and use of jasmonate for treating bladder dysfunction is provided.
BACKGROUND
Bladder Dysfunction
Bladder dysfunction is caused by abnormal functioning of the bladder tissue resulting in dysuria, irritative symptoms of urgency, frequency and nocturinia and the obstructive symptoms of reduced flow rate, incomplete emptying, hesitancy and increased time to urinate. One type of bladder dysfunction is an overactive bladder. This may be due to a number of factors.
One such cause of an overactive bladder is benign prostrate hyperplasia (BPH), a common condition in aging men. Approximately 80% of men over 50 years of age have varying degrees of urinary bladder outlet obstruction secondary to BPH (Levin et al., 1999, Urology 53:446-450). The bladder dysfunction symptoms are related to the effect of the enlarged prostrate on the urethra which creates a partial urethral obstruction.
The urethral obstruction caused by the enlarged prostrate initiates compensatory changes in the bladder including increased bladder wall thickness, decreased compliance, and detrusor denervation which are mediated by three cellular processes: 1) reduced nerve density; 2) mitochondrial dysfunction and 3) dysregulation of calcium storage and release from the sarcoplasmic reticulum involving Ca2+ATPase dysfunction (see, for example, US 2003/0187059).
Ca2+ATPase activities are approximately 40% lower in bladder samples of men with benign prostate hyperplasia (BPH) mediated bladder dysfunction as compared to Ca2+ATPase levels of bladder samples obtained from men of similar ages with no obstructive symptoms (Levin et al., 1999, Urology 53:446-450). The contractile dysfunctions associated with partial outlet obstruction correlate with disrupted sarcoplasmic reticulum (SR) Ca2+ storage release mechanisms and the degree of the contractile dysfunction is directly proportional to the degradation of sarcoplasmic endoplasmic reticulum calcium ATPase (SERCA) as shown using both enzymatic activity assays and Western blot analysis (see, for example, Zhao et al., 1997, Urology 49: 293-300).
Bladder dysfunction, particularly overactive bladder and more particularly, incontinence and poor detrusor contraction in women all increase as a function of age (reviewed in Li et al., 2009, Urology 74:691-697). A study presented by the Bladder Health Council of the American Foundation for Urologic Disease stated that incontinence affects more than 10 million Americans 85% who are women and affects over 20% of women over the age of 45 and the number increases with age.
In women, alterations in the female sex hormones are believed to play a major role in mediating symptoms of an overactive bladder such as including incontinence, urgency, and frequency due to observed weakness of urethral and pelvic floor muscles and underactive detrusuor smooth muscle contraction in low estrogen states. A possible explanation for the dysfunction is that there is a direct relationship between estrogen levels and blood flow to the bladder and urethral tissue which increases free radical generation and oxidative damage. The oxidative damage associated with estrogen levels as a result of reduced blood flow to the bladder and urethra could account for the decreased Ca2+ATPase activity associated with the bladder and the urethra in low estrogen states such as menopause. Total Ca2+ATPase activity of both the muscle and mucosa was significantly reduced in ovarectomized rabbits (OX) after 28 days indicating that calcium storage and release are compromised after OX (Li et al., 2009, Urology 74:691-697).
Other causes of outlet obstruction that results in bladder dysfunction include but are not limited to cancer, sclerosis or fibrosis of the bladder neck, urethral structure disease, urethral valves, and smooth and striated sphincter dyssynergia, (reviewed in US 2006/0234939). Furthermore, children may suffer from congenital urethral obstruction (U.S. Pat. No. 6,191,156).
An overactive bladder may also occur as a result of neurological damage due to disorders including but not limited to stroke, Parkinson's disease, diabetes, multiple sclerosis, peripheral neuropathy or spinal cord lesions.
Various therapies have been tried to treat bladder dyfunction, particularly overactive bladders. These include but are not limited to anticholinergic agents, prostaglandin inhibitors, beta-adrenergic agonists, COX2 inhibitors, calcium channel modulators (see, for example, U.S. Pat. Nos. 6,444,685, 6,19,156, 6,514,991, 7,459,430). Additionally, it has been observed that phospholamban, an inhibitor of SERCA is present in bladder sarcoplasmic reticulum and may play a role in bladder contractility (Nobe et al., 2001, J. Physiol. 535: 867-878).
Jasmonates
Jasmonates are a family of plant stress hormones that are found in minute quantities in edible plants and characterized by cyclopentone rings. Various uses for jasmonates have been disclosed. Examples include enhancing plant growth (U.S. Pat. No. 5,436,226), repelling insects (U.S. Pat. No. 5,118,711), treating cancer (U.S. Pat. No. 6,469,061) and treating skeletal muscle degeneration caused by malnutrition and disease (U.S. Pat. No. 6,465,021, US Patent Appln. Pub. No. 201000003346), pain relief (WO 2009019693), relieving psychological stress (US Patent Appln. Pub. No. 200700420567), use as a component of a sleep supplement (JP2000355545), treating dry skin (US Patent Appln. Pub. No. 20110085999), treating malodors on fabrics (US Patent Appln. Pub. No. 20110070181) treating heart failure and related disorders (US2011. Jasmonate has also been found to increase Ca2+Atpase in cardiac sarcoplasmic reticulum (see, for example, Antipenko et al., 1997, J. Biol. Chem. 272:2852-60) and skeletal muscle (see, for example, Ioumaa et al., 2002, J. Pharmacol. Exp. Ther. 300:638-46; Starling et al, 1995, Biochem. J. 308:343-6 and Starling et al., 1994, Biochemistry 15:3023-31).
SUMMARY
Provided is a method for modulating bladder contractility in a subject in need thereof comprising administering an amount of at least one jasmonate and optionally at least one other substance, wherein said substance is a drug or natural substance used to treat bladder dysfunction, effective to modulate said bladder contractility. In a related aspect, the invention relates to a method for treating bladder dysfunction, particularly, non-psychological stress related bladder dysfunction, even more particularly, an overactive bladder in a subject comprising administering an amount of at least one jasmonate effective to treat said bladder dysfunction, particularly, non-psychological stress related bladder dysfunction, even more particularly, an overactive bladder which even more particularly is an age-related overactive bladder and/or non-psychological stress related overactive bladder. In a particular embodiment, the subject is a mammal and in an even more particular embodiment, the subject is human.
Further provided, are compositions for modulating bladder contractility and/or non-psychological stress related bladder dysfunction in a mammal (e.g., human) comprising at least one jasmonate and optionally at least one other substance, wherein said substance may be a drug or natural substance used to treat said bladder dysfunction. The substance may be an antioxidant, a vitamin, amino acid and/or nutritional supplement. The composition may comprise jasmonate (e.g., methyl jasmonate), at least two basic amino acids (e.g., arginine and citrulline) and a nutritional supplement (e.g. taurine). The composition may comprise jasmonate, arginine, citrulline and taurine. In particular, the composition may comprise methyl jasmonate, arginine, citrulline and taurine or alternatively, cis-jasmone, arginine, citrulline and taurine.
In a related aspect, also provided is the use of jasmonate and optionally one other substance, wherein said substance is a drug or natural substance used to treat bladder dysfunction, particular an overactive bladder (e.g., age related overactive bladder), for formulating a medicament for use in modulating bladder contractility and/or for treating bladder dysfunction (e.g., non-psychological stress related bladder dysfunction), particularly an overactive bladder (e.g., age related overactive bladder, non-psychological stress related overactive bladder) in a mammal (e.g., human).


DETAILED DESCRIPTION OF SPECIFIC EMBODIMENTS
Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges is also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either both of those included limits are also included in the invention.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, the preferred methods and materials are now described.
It must be noted that as used herein and in the appended claims, the singular forms “a,” “and” and “the” include plural references unless the context clearly dictates otherwise.
It must be noted that as used herein and in the appended claims, the terms “composition” and “formulation” are used interchangeably.
DEFINITIONS
As defined herein, the term “modulate” means adjusting amount and/or rate of bladder contractility. Preferably, bladder contraction is stimulated.
As defined herein, the term an “overactive bladder” is any type of lower urinary tract disorder characterized by increased frequency of a desire to void, whether complete or episodic and where lost of voluntary control ranges form partial to total and where there is loss of urine (incontinence) or not. Symptoms include, but are not limited to, urinary urgency, incontinence, urge incontinence, urinary frequency and nocturia.
Jasmonates
The jasmonates used in the compositions and methods disclosed herein may have the formula I




wherein:

    
    
        n is 0, 1, or 2;
        R1 is OH, alkoxy, O-glucosyl, or imino,
        R2 is OH, O, alkoxy, or O-glucosyl,
        R3, R4, and R5 are H, OH, alkoxy or O-glucosyl,
        and/or wherein R1 and R2, or R1 and R4 together form a lactone, and further wherein the bonds between C3:C7, C4:C5, and C9:C10 may be double or single bonds; or a derivative of said formula, wherein the derivative has at least one of the following:
        
            a lower acyl side chain at C3 (free acid or ester or conjugate), a keto or hydroxy (free hydroxy or ester) moiety at the C6 carbon, or an n-pentenyl or n-pentyl side chain at C7.
        
        
    
    


In a particular embodiment, the jasmonate may be at least one member selected from the group consisting of methyl jasmonate, jasmonic acid, jasmone, 7-iso-jasmonic acid, 9,10-dihydrojasmonic acid, 2,3-didehydrojasmonic acid, 3,4-didehydrojasmonic acid, 3,7-didehydrojasmonic acid, 4,5-didehydrojasmonic acid, 4,5-didehydro-7-iso-jasmonic acid, cucurbic acid, 6-epi-cucurbic acid, 6-epi-cucurbic-acid lactone, 12-hydroxy-jasmonic acid, 12-hydroxy-jasmonic-acid-lactone, 11-hydroxy-jasmonic acid, 8-hydroxy-jasmonic acid, homo-jasmonic acid, dihomo-jasmonic acid, 11-hydroxy-dihomo-jasmonic acid, 8-hydroxy-dihomo-jasmonic acid, tuberonic acid, tuberonic acid-O-β-glucopyranoside, cucurbic acid-O-β-glucopyranoside 5,6-didehydrojasmonic acid, 6,7-didehydro-jasmonic acid, 7,8-didehydrojasmonic acid, cis-jasmone, methyl-dihydro-isojasmonate, dihydro-jasmone, amino acid conjugates of jasmonic acid, the lower alkyl esters of said jasmonic acids, and the carrier ligand conjugates and the sterioisomers thereof.
Compositions
The compositions may comprise the jasmonate set forth above. Additionally, the compositions may further comprise least one other drug or natural substance used to modulate bladder contractility and/or treat bladder dysfunction, particularly non-stress related bladder dysfunction, even more particularly, an overactive bladder, even yet more particularly, age-related overactive bladder. This drug or substance may include but is not limited to substances that will be effective in treating bladder dysfunction and particularly an overactive bladder and/or that potentiates the effect of jasmonate. Such substances may include but are not limited to duloxetine, monoamine reuptake inhibitors, spasmolytics, anticholinergics, beta-3 adrenergic receptor agonists (see, for U.S. Pat. Nos. 6,444,685 and 6,569,873), calcium sensitizers, antioxidants (see, for example, US 2003/0187059), calcium channel modulators (see, for example, US 2006/0234939).
The compositions may also include jasmonates in combination with a drug or substance that may include but is not limited to Astragalus or substances derived therefrom (e.g., astragaloside), gingerol, taurine, green tea or substances derived therefrom (e.g., epigallocatechin gallate), gingerol, amino acids such as taurine and arginine and citrulline. In a particular embodiment, the composition may comprise methyl jasmonate, arginine, citrulline and taurine.
The compositions, in particular, pharmaceutical compositions can be formulated for administration by a variety of routes including oral, transdermal, parenteral (subcutaneous, intraperitoneal, intravenous, intra-arterial, and intramuscular). The compositions may also be formulated for transurethral administration and may be in the form of in the form of a suppository (see, for example, US 2006/0234939). Alternatively, the compositions may be formulated for vaginal administration and may be in the form of vaginal suppositories, creams, ointments, liquid formulations, pessaries, tampons, gels, pastes, foams or sprays (See, for example, US 2006/0234939).
Such compositions are prepared in a manner well known in the pharmaceutical art and comprise as an active ingredient at least one of the compounds used in the methods as described herein above and a pharmaceutically acceptable excipients or a carrier. The amount of the active ingredient(s) in the composition of the present invention is from about 0.5 to 100% per weight. The term “pharmaceutically acceptable” means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals and, more particularly, in humans. Choosing suitable ingredients for the composition is a routine for those of ordinary skill in the art. It is evident that suitable carriers, solvents, gel forming ingredients, dispersion forming ingredients, antioxidants, colors, sweeteners, wetting compounds, release controlling components and other ingredients normally used in this field of technology may be also used. Pharmaceutical compositions in the form of intravenous solutions are preferred. The active ingredients may be formulated in the same pharmaceutical formulation. Alternatively, the active ingredients are formulated as separate pharmaceutical dosage forms. The combination of the pharmaceutical dosage forms may be packaged as a single medical product or kit for use in the method of the invention, optionally together with a package insert instructing to the correct use of the medical product.
Another preferred embodiment of the invention is a medical product comprising, separately or together, as active ingredients one or more jasmonates or a pharmaceutically acceptable salt thereof and one or more other substance used for reducing muscle fatigue and/or increasing skeletal muscle performance in a mammal as a combined preparation.
In yet another embodiment, the medical product may comprise one two or more jasmonates set forth above in separate compositions.
Administration and Uses
As noted above, one or more jasmonate(s) optionally in combination with other substances may be used to treat bladder dysfunction, particularly, non-stress related bladder dysfunction. In a specific embodiment, the jasmonate optionally in combination with other substances may be used to treat an overactive bladder, in particular a non-stress related overactive bladder. Non-psychological stress related overactive bladder includes but is not limited to age-related overactive bladder or neurogenic overactive bladder. An age-related overactive bladder may in particular embodiment, be due to prostate hyperplasia, a disorder associated with benign enlargement of the prostate gland or in another embodiment an estrogen deficiency associated with menopause. A neurogenic overactive bladder may in one embodiment occur as a result of neurological damage due to disorders including but not limited to stroke, Parkinson's disease, diabetes, multiple sclerosis, peripheral neuropathy or spinal cord lesions.
The active ingredients may be administered simultaneously, separately or sequentially. The administration routes of the active ingredients include, but are not limited to, enteral, e.g. oral or rectal, or parenteral, e.g. intravenous, intramuscular, transurethral, vaginal, intraperitoneal or transdermal.
The active ingredients furthermore may be administered as an immediate release formulation (a drug formulation that provides for release of the drug immediately after drug administration) controlled release formulation (a formulation in which release is not immediate) or a sustained release formulation (a drug formulation that provides for gradual release of a drug over an extended period of time, and that preferably, although not necessarily, results in substantially constant blood levels of a drug over an extended time period such as up to about 72 hours, about 66 hours, about 60 hours, about 54 hours, about 48 hours, about 42 hours, about 36 hours, about 30 hours, about 24 hours, about 18 hours, about 12 hours, about 10 hours, about 8 hours, about 4 hours, after drug administration). In a particular embodiment, the compositions may be administered prior to commencement of an activity where suppression of symptoms of an overactive bladder would be desirable.
The compositions used may preferably be administered orally preferably once per day. According to the invention, the suggested daily dose of jasmonate(s) is in general from about 0.01 to 50 mg, preferably from about 0.02 to 20 mg, more preferably from about 0.05 to 10 mg, and even more preferably, from about 0.05 mg-0.10 mg, depending on the age, body weight and condition of the patient. The effective amount of jasmonate(s) to be administered to a subject depends upon the condition to be treated, the route of administration, age, weight and the condition of the patient. Similar dosages of other substances may also be used.
EXAMPLES
The composition and methods set forth above will be further illustrated in the following, non-limiting Examples. The examples are illustrative of various embodiments only and do not limit the claimed invention regarding the materials, conditions, weight ratios, process parameters and the like recited herein.
Five subjects, 4 male and 1 female agreed to participate in the study. Subjects ranged in age from 55-73 with average age being 67. All participants experienced noticeable urinary bladder dysfunction symptoms including frequency, urgency, and reduced urine flow. Subjects took from 50 to 100 micrograms of Jasmone once a day for 30 days. At two weeks approximately 1⁄2 of the subjects experienced improved bladder function. By day thirty, all subjects reported improvements in frequency, urgency, and urine flow. For example, several subjects noted that before treatment they averaged 2 night time awakenings to urinate which was reduced to 1 or 0 subsequent to taking Jasmone. Other subjects had even greater improvement from an average of 3-4 night awakenings to urinate to 1. Daytime bathroom visits were also reduced approximately 50%. Several patients also noted radically reduced urgency, which increased “holding time” from approximately 10 minutes to 60 minutes i.e. during extended air travel. In addition several male subjects reported increased flow of discharge up to 100%.
This invention may be embodied in other forms or carried out in other ways without departing from the spirit or essential characteristics thereof. The present disclosure is therefore to be considered as in all aspects illustrate and not restrictive, and all changes which come within the meaning and range of equivalency are intended to be embraced therein.
Various references are cited throughout this specification, each of which is incorporated herein by reference in its entirety.

## Claims

1. A method for treating a neurogenic overactive bladder in a mammal in need thereof comprising administering an amount of at least one jasmonate effective to treat said neurogenic overactive bladder.

2. The method according to claim 1, wherein said jasmonate has the structure (I)




wherein:
n is 0, 1, or 2;
R1 is OH, alkoxy, O-glucosyl, or imino,
R2 is OH, O, alkoxy, or O-glucosyl,
R3, R4, and R5 are H, OH, alkoxy or O-glucosyl,
and/or wherein R1 and R2, or R1 and R4 together form a lactone, and further wherein the bonds between C3:C7, C4:C5, and C9:C10 may be double or single bonds; or a derivative of said structure, wherein said derivative is a jasmonate of structure I modified by one or more substituents selected from the group consisting of a lower acyl free acid, ester or conjugate side chain at C3, a keto, hydroxy and ester moiety at the C6 carbon, or an n-pentenyl or n-pentyl side chain at C7.

3. The method according to claim 2, wherein said jasmonate is a compound selected from the group consisting of methyl jasmonate, jasmonic acid, jasmone, 7-iso-jasmonic acid, 9,10-dihydrojasmonic acid, 2,3-didehydrojasmonic acid, 3,4-didehydrojasmonic acid, 3,7-didehydrojasmonic acid, 4,5-didehydrojasmonic acid, 4,5-didehydro-7-iso-jasmonic acid, cucurbic acid, 6-epi-cucurbic acid, 6-epi-cucurbic-acid lactone, 12-hydroxy-jasmonic acid, 12-hydroxy-jasmonic-acid-lactone, 11-hydroxy-jasmonic acid, 8-hydroxy-jasmonic acid, homo-jasmonic acid, dihomo-jasmonic acid, 11-hydroxy-dihomo-jasmonic acid, 8-hydroxy-dihomo-jasmonic acid, tuberonic acid, 5,6-didehydrojasmonic acid, 6,7-didehydro-jasmonic acid, 7,8-didehydrojasmonic acid, cis-jasmone, methyl-dihydro-isojasmonate, dihydro-jasmone, amino acid conjugates of jasmonic acid, the lower alkyl esters of said jasmonic acids, and the stereoisomers thereof.

4. The method according to claim 1, which further comprises administering at least one other substance, wherein said substance is a drug or natural substance, wherein said drug or natural substance is used to treat an overactive bladder.

5. The method to claim 4, wherein said substance is a drug or natural substance, wherein said drug or natural substance is an antioxidant, a vitamin, amino acid, nutritional supplement, or combination of the foregoing.

6. The method according to claim 4, wherein jasmonate, citrulline, arginine and taurine are administered to said mammal.

7. A method for modulating bladder contractility and/or treating non-psychological stress-related bladder dysfunction in a female mammal in need thereof comprising administering an amount of at least one jasmonate effective to modulate said bladder contractility and/or to treat said non-psychological stress related bladder dysfunction.

8. The method according to claim 7, wherein said non-psychological stress related bladder dysfunction is an age-related overactive bladder.

9. The method according to claim 8, wherein said age-related overactive bladder is due to estrogen deficiency associated with menopause.

10. The method according to claim 7, wherein said female mammal is a human woman.

11. The method according to claim 7, wherein said jasmonate has the structure (I)




wherein:
n is 0, 1, or 2;
R1 is OH, alkoxy, O-glucosyl, or imino,
R2 is OH, O, alkoxy, or O-glucosyl,
R3, R4, and R5 are H, OH, alkoxy or O-glucosyl,
and/or wherein R1 and R2, or R1 and R4 together form a lactone, and further wherein the bonds between C3:C7, C4:C5, and C9:C10 may be double or single bonds; or a derivative of said structure, wherein said derivative is a jasmonate of stricture I modified by one or more substituents selected from the group consisting of a lower acyl free acid, ester or conjugate side chain at C3, a keto, hydroxy or ester moiety at the C6 carbon, or an n-pentenyl or n-pentyl side chain at C7.

12. The method according to claim 7, wherein said jasmonate is a compound selected from the group consisting of methyl jasmonate, jasmonic acid, jasmone, 7-iso-jasmonic acid, 9,10-dihydrojasmonic acid, 2,3-didehydrojasmonic acid, 3,4-didehydrojasmonic acid, 3,7-didehydrojasmonic acid, 4,5-didehydrojasmonic acid, 4,5-didehydro-7-iso-jasmonic acid, cucurbic acid, 6-epi-cucurbic acid, 6-epi-cucurbic-acid lactone, 12-hydroxy-jasmonic acid, 12-hydroxy-jasmonic-acid-lactone, 11-hydroxy-jasmonic acid, 8-hydroxy-jasmonic acid, homo-jasmonic acid, dihomo-jasmonic acid, 11-hydroxy-dihomo-jasmonic acid, 8-hydroxy-dihomo-jasmonic acid, tuberonic acid, 5,6-didehydrojasmonic acid, 6,7-didehydro-jasmonic acid, 7,8-didehydrojasmonic acid, cis-jasmone, methyl-dihydro-isojasmonate, dihydro-jasmone, amino acid conjugates of jasmonic acid, the lower alkyl esters of said jasmonic acids, and the stereoisomers thereof.

13. The method according to claim 7, which further comprises administering at least one other substance, wherein said substance is a drug or natural substance, wherein said drug or natural substance is used to modulate bladder contractility and/or treat bladder dysfunction.

14. The method to claim 13, wherein said substance is a drug or natural substance, wherein said drug or natural substance is an antioxidant, a vitamin, amino acid, nutritional supplement, or combination of the foregoing.

15. The method according to claim 13, wherein jasmonate, citrulline, arginine and taurine are administered to said mammal.

16. The method according to claim 1, wherein said jamonate is selected from the group consisting of tuberonic acid-O-β-glucopyranoside and cucurbic acid-O-β-glucopyranoside.

17. The method according to claim 7, wherein said jamonate is selected from the group consisting of tuberonic acid-O-β-glucopyranoside and cucurbic acid-O-β-glucopyranoside.