COMPOSITIONS AND METHODS FOR TREATING ERECTILE

DYSFUNCTION

[001] The present invention provides combined compositions and kits comprising Isosorbide mono-nitrate, dimethyl Isosorbide (DMI) or Isosorbide dinitrate, Hydralazine, Moxisylyte, Arginine, Norvaline, or any combination thereof, and a topical carrier. Methods for treating erectile dysfunction, relieving erectile dysfunction symptoms, or treating female sexual arousal disorder utilizing the compositions and kits of the invention are also provided.

BACKGROUND OF THE INVENTION

[002] More than 18 million men over the age of 20 in the U.S. are affected by erectile dysfunction (ED) or insufficient erection. Penile erection is mainly mediated by the parasympathetic nervous system, which when stimulated causes arterial dilation and relaxation of the cavernosal smooth muscle. The increased blood flow into the Corpora Cavernosa, together with reduced venous outflow, results in penile rigidity and erection.

[003] ED, insufficient erection or Female sexual arousal disorder (FSAD) are sexual dysfunctions. There are no universally agreed criteria for how consistent the problem has to be and for what duration it needs to be present to fulfill the definition. A period of persistence for longer than 3 months has been suggested as a reasonable clinical guideline.

[004] In the past, patients have underreported this problem because of embarrassment and the belief that little could be done to alleviate it. The availability and marketing of new therapies for ED have greatly increased public awareness of this problem.

[005] In several studies, it was clearly shown that ED is correlated to age. Generally, saying, fewer than 2% of the men who reported that they had erection problems experienced them before age 40, and 4% had experienced problems between age 40 and 49. From age 50 upwards, the percentage of men reporting ED increased dramatically with 26% between the ages of 50 to 59, 40% aged 60 to 69 years and 61% for men older than 70 having experienced ED.

[006] Erectile function is a hemodynamic process of blood in-flow and pressure maintenance in the cavernosal spaces. Christ, Urol. Clin. North Am., 22: 727 (1995). Following sexual arousal and the release of nitric oxide (NO) to the erectile tissue, three processes occur to achieve an erection. These are relaxation of the trabecular smooth muscle, arterial dilation and venous compression. During the final stage, arterial flow fills sinusoidal spaces, compressing subtunical venules thereby reducing venous outflow. Blood flows into the cavernous spaces of the penis, thus expanding and stretching the penis into a rigid organ. The flow of blood in and out of the cavernous spaces is controlled by cavernous smooth muscle cells embedded in the trabeculae of the cavernous spaces. With normal erectile function, a high intracavernous pressure (ICP) is maintained with a low inflow rate. Karadeniz, et al., Urol. Int., 57: 85 (1996).

[007] The penis is a predominantly vascular organ, and vascular or penile arterial insufficiency is the most common etiology of erectile dysfunction. Sinusoidal smooth muscle atrophy and collagen deposition is a common finding in men with long standing ED of various etiologies, whether due to hormonal, neurological or vascular causes. Karadeniz, et al., Urol. Int., 57: 58 (1996). Such degradation in smooth muscle quantity and quality leads to veno-occlusive dysfunction. This represents an end-stage muscular degeneration akin to myocardial changes with congestive heart failure or dilated cardiomyopathy for which no treatment currently exists with hope of reversing the underlying pathologic process.

[008] Veno-occlusive disease is a common finding among patients with ED. Following radical prostatectomy, for example, approximately 30% of patients may have vasculogenic ED in addition to neurogenic ED and at least half of these men may have venous leak. Regardless of the etiology of organic ED (neurogenic, traumatic, hormonal, and vascular, etc.), venous leakage is a common final condition resulting from smooth muscle atrophy. Mersdorf, et al., J. Urol., 154: 749 (1991). Veno-occlusive dysfunction is the most common etiology of ED among non-responders to medical management of ED. None of the medical therapy currently exists is curative for this condition. Patients with veno-occlusive dysfunction exhibit a poor response to intracavernous injection with vasoactive agents (papavarine, prostaglandin El, phentolamine, or combinations, for example), despite good arterial flow demonstrated by duplex ultrasound. The diagnosis of veno-occlusive disease may be confirmed with specific findings on cavernosometry and cavernosography. Nehra, et al., J. Urol., 156: 1320 (1996).

[009] Atherosclerotic or traumatic arterial occlusive disease of the pudendal-cavernous- helicine arterial tree can decrease the perfusion pressure and arterial flow to the sinusoidal spaces, thus decreasing the rigidity of the erect penis. Common risk factors associated with generalized arterial insufficiency include hypertension, hyperlipidemia, cigarette smoking, diabetes mellitus, and pelvic irradiation. Goldstein, et al., JAMA, 251: 903-910 (1984), Rosen, M. P., et al., Radiology, 174(3 Pt 2): 1043-48 (1990), Levine, F. J., et al., J. Urology, 144(5): 1147-53 (1990). Epidemiological studies have shown a high incidence of ED in patients with coronary arterial disease. Heaton, J. P., et al., Int'l J. Impotence Res., 8(1): 35- 39 (1996). Focal lesion of the common penile or cavernous artery is most often seen in young patients who have sustained blunt pelvic or perineal trauma such as in cases of biking accidents. Levine, F. J., et al., J. Urology, 144(5): 1147-53 (1990).

[0010] Because of the close proximity of the cavernous nerves to the capsule of the prostate,

ED is a frequent complication after radical prostatectomy or cystectomy and prostatic cryosurgery. Although the nerve-sparing prostatectomy technique developed by Walsh, et al., Br. J. Urol., 56: 694 (1984) has significantly reduced the postoperative impotence rate, a large number of patients still suffer from inadequate penile rigidity. Peripheral nerve regeneration is a slow process, and the fact that most patients do not recover potency for 6 months to 2 years indicates substantial axonal damage, even with preservation of the neural sheath. An anatomic study of the cavernous nerves by Paick et al., Urology, 42: 145 (1993) revealed both a medial and a lateral bundle of cavernous nerves at the level of the prostate, suggesting that in some cases the lateral bundle can be saved, even in non-nerve- sparing prostatectomy.

[0011] Stimulation of penile shaft by the nervous system leads to the secretion of nitric oxide (NO), which causes the relaxation of smooth muscles of Corpora Cavernosa, and subsequently penile erection. Importantly, there are several mechanisms by which NO release can be induced.

[0012] The current nonsurgical treatments that are not device based include: oral therapies, intracavernous injection therapies, and intraurethral therapies. Specifically, oral therapy has revolutionized the treatment approach to patients with ED. Three oral agents were approved for use by the FDA: Sildenafil (Viagra), vardenafil (Levitra) and tadalafil (Cialis). However, these three medications are associated with multiple side effects.

[0013] Female sexual arousal disorder (FSAD) refers to the persistent or recurrent inability of a woman to achieve or maintain an adequate lubrication-swelling response during sexual activity. This lack of physical response may be either lifelong or acquired, and either generalized or situation- specific. FSAD has both physiological and psychological causes. The results of FSAD are often sexual avoidance, painful intercourse, and sexual tension in relationships.

[0014] There are currently no FDA-approved topical drugs for ED. The literature describes experimental or anecdotal use of topical preparations containing alprostadil, papaverine, combinations of aminophylline, Isosorbide dinitrate (or dimethyl Isosorbide (DMI)) and co- dergocrine mesylate, NO nanoparticles, L-Arginine and testosterone; however, such preparations did not reach the market place presumably due to limited efficacy, side effects and insufficient scientific background.

FIGURES

[0015] Figure 1. Is a bar graph showing the cumulative amount of drug permeated across human penile epidermal membrane after t=6 hours (mean + SEM for n=4-6 replicates) for single drug systems.

[0016] Figure 2. Is a bar graph showing the cumulative amount of drug permeated across human penile epidermal membrane after t=24 hours (mean + SEM for n=4-6 replicates) for single drug systems.

[0017] Figure 3. Is a bar graph summarizing the calculated putative efficacy index for individual drugs solutions with and without a penetration enhancer.

[0018] Figure 4. Is a graph showing the cumulative amount of drug permeated across human penile epidermal membrane after t=24 h (mean + SEM for n=4-6 replicates), for single drug saturated solutions only.

[0019] Figure 5. Is a graph showing the cumulative amount of ISDN permeated across human penile epidermal membrane after t=24 hours (mean + SEM for n=4-6 replicates).

[0020] Figure 6. A graph showing blood flow as a consequence of treating mice with the following formulations: 35/36: ISDN (4%, 3%) + Hydralazine (2%, 1.5%) + DMI based formulations; 37: ISDN + Hydralazine + Moxisylyte (3%, 1.5%, 0.5%) + DMI based formulation; 38: Phetolamine (1%) + DMI based formulation.

[0021] Figure 7. A graph showing vessel diameter as a consequence of treating mice with the following formulations: 35/36: ISDN (4%,3%) + Hydralazine (2%, 1.5%) + DMI based formulations; 37: ISDN + Hydralazine + Moxisylyte (3%, 1.5%, 0.5%) + DMI based formulation; 38: Phetolamine (1%) + DMI based formulation. [0022] Figure 8. A graph showing mean cumulative amount of ISDN released per unit area across PTFE membrane over a 30 hours (h) period of time (Mean + SD, n=6 per formulation with the exception of FIA5, SSH1, SSH3, SSH4 and SSH8, where n=5 is reported).

[0023] Figure 9. A graph showing mean cumulative amount of Moxisylyte released per unit area across PTFE membrane over a 30 h period of time (Mean + SD, n=6 per formulation with the exception of FIA5, where n=5 is reported).

[0024] Figure 10. A graph showing change % in Blood Flow following treatment with different combinations of selected formulations (table 17; number 38). Statistical analysis for blood flow *=p<0.05; **= p<0.01 and ***=p<0.001 compared to vehicle.

[0025] Figure 11. A graph showing change % in Blood Vessel Diameter following treatment with different combinations of selected formulations (table 17; number 38).

Statistical analysis for blood flow *=p<0.05; **= p<0.01 and ***=p<0.001 compared to vehicle.

[0026] Figure 12. A graph showing change % in Blood Flow following treatment with VL#FIA3-30. Statistical analysis for blood flow *=p<0.05; **= p<0.01 and ***=p<0.001 compared to vehicle.

[0027] Figure 13. A graph showing change % in Blood Vessel Diameter following treatment with VL#FIA3-30. Statistical analysis for blood flow *=p<0.05; **= p<0.01 and ***=p<0.001 compared to vehicle.

[0028] Figure 14. A graph showing the correlation between blood flow and vessels' diameter in individual measurements.

SUMMARY OF THE INVENTION

[0029] In one embodiment, this invention provides a composition comprising Isosorbide mono-nitrate, dimethyl Isosorbide or Isosorbide dinitrate, a nitric oxide donor, and a topical carrier. [0030] In one embodiment, this invention further provides a composition comprising Isosorbide mono-nitrate, dimethyl Isosorbide, or Isosorbide dinitrate, Hydralazine or a salt thereof, and a topical carrier.

[0031] In one embodiment, this invention further provides a composition comprising Isosorbide mono-nitrate, dimethyl Isosorbide, or Isosorbide dinitrate, Arginine, and a topical carrier.

[0032] In one embodiment, this invention further provides a composition comprising Isosorbide mono-nitrate, dimethyl Isosorbide, or Isosorbide dinitrate, Moxisylyte or a salt thereof, and a topical carrier. In one embodiment, this invention further provides a composition comprising Arginine, Hydralazine or a salt thereof, and a topical carrier.

[0033] In one embodiment, this invention further provides a kit comprising a first composition and a second composition, wherein the first composition is a topical composition comprising Isosorbide mono-nitrate, dimethyl Isosorbide, Isosorbide dinitrate, or a combination thereof, the second composition is a powder comprising Moxisylyte, a Moxisylyte salt, Hydralazine, a Hydralazine salt, or any combination thereof, wherein the first composition and the second composition are physically separated. In one embodiment, the kit further comprises mixing and/or dosing instructions of for the first composition and the second composition. In one embodiment, the second composition is a dry powder.

[0034] In one embodiment, this invention further provides a method for inducing a penile erection in a subject, comprising the step of administering to the subject a composition comprising an effective amount of: Isosorbide mono-nitrate, dimethyl Isosorbide, or Isosorbide dinitrate, a nitric oxide donor, and a pharmaceutical acceptable carrier, thereby inducing erection in a subject. In one embodiment, inducing penile erection is prolonging penile erection. [0035] In one embodiment, this invention further provides a method for treating erectile dysfunction or relief erectile dysfunction symptoms or female sexual arousal disorder in a subject, comprising the step of administering to the subject a composition comprising an effective amount of: Isosorbide mono-nitrate, dimethyl Isosorbide, or Isosorbide dinitrate, a nitric oxide donor, and a pharmaceutical acceptable carrier, thereby treating erectile dysfunction or relief erectile dysfunction symptoms or female sexual arousal disorder in a subject.

[0036] In one embodiment, this invention further provides a method for inducing penile erection in a subject, comprising the step of administering to the subject a composition comprising an effective amount of: Arginine, Hydralazine or a salt thereof, Moxisylyte, Norvaline, or any combination thereof and a pharmaceutical acceptable carrier, thereby inducing a penile erection in a subject.

[0037] In one embodiment, this invention further provides a method of treating erectile dysfunction or relief erectile dysfunction symptoms or female sexual arousal disorder in a subject, comprising the step of administering to the subject a composition comprising an effective amount of: Arginine, Hydralazine or a salt thereof and a pharmaceutical acceptable carrier, thereby treating erectile dysfunction or relief erectile dysfunction symptoms or female sexual arousal disorder in a subject.

DETAILED DESCRIPTION OF THE PRESENT INVENTION

[0038] In the following detailed description, numerous specific details are set forth in order to provide a thorough understanding of the invention. However, it will be understood by those skilled in the art that the present invention may be practiced without these specific details. In other instances, well-known methods, procedures, and components have not been described in detail so as not to obscure the present invention. [0039] This invention provides, in one embodiment, a composition comprising Isosorbide mono-nitrate, dimethyl Isosorbide, or Isosorbide dinitrate, a nitric oxide donor, and a topical carrier. In another embodiment, a composition described herein is a topical composition. In another embodiment, a composition described herein comprises a topical carrier which enables the penetration of the combined active pharmaceutical ingredients (APIs) described herein into a skin layer. In another embodiment, a composition described herein comprises a topical carrier which enables the penetration of the combined active pharmaceutical ingredients (APIs) described herein to the Corpus Cavernosum, Tunica Albugines, Cavernosal Space, Cavernosal Artery, or any combination thereof. In another embodiment, the composition described herein, when applied topically to the penis, enables the penetration of an effective amount of the combined active pharmaceutical ingredients (APIs) to the Corpus Cavernosum penis, Corpus Cavernosum clitoridis, Tunica Albugines, Cavernosal Space, Cavernosal Artery, or any combination thereof.

[0040] In another embodiment, the invention provides a kit comprising a first composition and a second composition, wherein the first composition is a topical composition comprising Isosorbide mono-nitrate, dimethyl Isosorbide, Isosorbide dinitrate, or a combination thereof, the second composition is a dry powder comprising Moxisylyte, a Moxisylyte salt, Hydralazine, a Hydralazine salt, or any combination thereof, wherein the first composition and the second composition are physically separated within the kit. In another embodiment, the first composition further comprises Arginine, L-Norvaline, or both. In another embodiment, the second composition further comprises Arginine, L-Norvaline, or both.

[0041] In another embodiment, "Moxisylyte" comprises Moxisylyte hydrochloride. In another embodiment, "Moxisylyte powder" comprises Moxisylyte hydrochloride powder. In another embodiment, Moxisylyte hydrochloride powder or Hydralazine (or a Hydralazine salt) powder comprises a crystalline powder. In another embodiment, Moxisylyte hydrochloride powder or Hydralazine (or a Hydralazine salt) comprises at least 50% Moxisylyte hydrochloride. In another embodiment, Moxisylyte hydrochloride powder or Hydralazine (or a Hydralazine salt) comprises at least 60% Moxisylyte hydrochloride. In another embodiment, Moxisylyte hydrochloride powder or Hydralazine (or a Hydralazine salt) comprises at least 70% Moxisylyte hydrochloride. In another embodiment, Moxisylyte hydrochloride powder or Hydralazine (or a Hydralazine salt) comprises at least 80% Moxisylyte hydrochloride. In another embodiment, Moxisylyte hydrochloride powder or Hydralazine (or a Hydralazine salt) comprises at least 90% Moxisylyte hydrochloride. In another embodiment, Moxisylyte hydrochloride powder or Hydralazine (or a Hydralazine salt) comprises at least 95% Moxisylyte hydrochloride.

[0042] In another embodiment, a composition as described herein comprises about 1 % w/w Moxisylyte. In another embodiment, a composition as described herein comprises about 0.2 to 2 % w/w Moxisylyte. In another embodiment, a composition as described herein comprises about 0.1 to 10 % w/w Moxisylyte.

[0043] In another embodiment, a composition as described herein ensures the dissolution time of less than 5 minutes. In another embodiment, a composition as described herein ensures the dissolution time of less than 3 minutes. In another embodiment, a composition as described herein ensures the dissolution time of less than 1 minute. In another embodiment, a composition as described herein ensures the dissolution time of 30 seconds to 1 minute. In another embodiment, a composition as described herein ensures the dissolution time of 15 seconds to 1 minute. In another embodiment, a composition as described herein ensures the dissolution time of 30 seconds to 45 seconds. In another embodiment, a composition as described herein ensures the dissolution time about 30 seconds. In another embodiment, the term "about" refers to +/- 20%.

[0044] In another embodiment, the w/w % ratio of ISDN and Moxisylyte in a composition as described herein is between 6: 1 and 1:4. In another embodiment, the w/w % ratio of ISDN and Moxisylyte in a composition as described herein is between 4:1 and 1:2. In another embodiment, the w/w % ratio of ISDN and Moxisylyte in a composition as described herein is between 3:1 and 1:1. In another embodiment, the w/w % ratio of ISDN and Moxisylyte in a composition as described herein is between 3:1 and 1:1. In another embodiment, the w/w % ratio of ISDN and Moxisylyte in a composition as described herein is between 2:1 and 1:1.

[0045] In another embodiment, ISDN is provided with lactose at a ratio (ISDN/Lactose) between 2:1 to 1:4. In another embodiment, ISDN is provided with lactose at a ratio of (ISDN/Lactose) between 1:1 to 1:4. In another embodiment, ISDN is provided with lactose at a ratio of (ISDN/Lactose) between 1: 1 to 1:2. In another embodiment, ISDN is provided with lactose at a ratio of (ISDN/Lactose) between 1:1 to 1:2. In another embodiment, ISDN is provided with lactose at a ratio of (ISDN/Lactose) about 2:3.

[0046] In another embodiment, a powder comprises an active ingredient as described herein and at least one excipient. In another embodiment, any powder excipient for topical use can be used according to the invention. In another embodiment, the powder excipient promotes dissolution, gelling, viscosity, penetration, release profile, or any combination thereof of a mixture comprising both the first and the second composition when applied to a body surface. In another embodiment, the excipient is a bulking agent. In another embodiment, the excipient is a moisture absorbing agent. In another embodiment, the excipient is a preservative. In another embodiment, a therapeutic composition of the invention comprises a deep drug/API penetration agent.

[0047] In another embodiment, the invention provides a kit as the actual combination of the active ingredients described herein is unstable. In another embodiment, the invention provides a kit as the actual combined preparations of the active ingredients described herein is unstable. In another embodiment, the invention provides a kit for storing each of the active ingredients separately and mixing them only immediately prior to use.

[0048] In another embodiment, the kit of the present invention comprises mixing instructions of the first composition and the second composition. In another embodiment, the kit of the present invention comprises a product insert which includes instruction pertinent to the topical utilization of the kit constituents including mixing instructions of the first composition and the second composition. In another embodiment, the kit of the present invention further comprises means for mixing the first composition and the second composition.

[0049] In another embodiment, a kit or a composition of the invention comprises a combination of: NO donor, activator of guanylate cyclase (GC) and a-blocking agent. In another embodiment, the kit of the present invention is aimed at physically separating an alpha-blocking agent such as Moxisylyte and NO donor such as dimethyl Isosorbide or Isosorbide dinitrate. In another embodiment, the kit of the present invention is aimed at physically separating an alpha-blocking agent such as Moxisylyte and guanylate cyclase activator such as Isosorbide dinitrate or dimethyl Isosorbide. In another embodiment, the kit of the present invention is aimed at physically separating an alpha-blocking agent such as Moxisylyte and a compound capable of acting both as a NO donor and a guanylate cyclase activator such as Isosorbide dinitrate or dimethyl Isosorbide. In another embodiment, the kit of the present invention is aimed at physically separating Moxisylyte and Isosorbide dinitrate or dimethyl Isosorbide. In another embodiment, the kit is aimed at physically separating agents that increase adenylate cyclase and cAMP activity such as Hydralazine; and 2) Norvaline is an inhibitor of arginase or L-arginase.

[0050] In another embodiment, it was unexpectedly discovered that separating the active ingredients described herein prior to use, dramatically enhances their combined therapeutic effect. In another embodiment, it was unexpectedly discovered that separating the active ingredients described herein prior to use, dramatically enhances their combined synergistic therapeutic effect. In another embodiment, it was unexpectedly discovered that separating the active ingredients overcomes dramatic instability drawback.

[0051] In another embodiment, a topical composition comprises a topical carrier as described herein. In another embodiment, the present invention involves therapeutically applying the recently mixed first composition and second composition. In another embodiment, the first composition and the second composition are manually applied as premeasured dosages or layers. In some embodiments, the first composition and the second composition are simply mixed just prior to application on a body surface to form a relative homogeneous mixture. In another embodiment, therapeutically applying is topically applying the combination to a body site. In some embodiments, the first composition and the second composition are mixed on a body surface such as the skin.

[0052] In another embodiment, extended release of an active agent as described herein is desired. In another embodiment, extended release can be a measure of viscosity or gelation of the topical composition or the combination of the topical composition and the powder.

[0053] In another embodiment, the compositions and methods described herein are combined with known ED and/or FSAD therapies and API such as but not limited to: PG El, PG El/prazosin, VIP/phentolamine, Forskolin, Nitroglycerine, Minoxidil, Papaverine, Milrinone, Sildenafil, IC351, Vardenafil, Phentolamine, Prazosin, Chlorpromazine, Yohimbine, Trazodone, Apomorphine, or any combinations thereof.

[0054] In another embodiment, the kit includes a multi-unit dose of each of the first and second compositions wherein each of the multi-unit dosage of each of the first and second compositions is housed in a physically separated compartment. In another embodiment, the kit includes a single unit dose pharmaceutical delivery system comprising two physically separated compartments. In another embodiment, the kit includes a single unit dose pharmaceutical delivery system comprising two physically separated compartments and an element adapted to connect the two compartments upon actuation. In another embodiment, the element adapted to connect the two compartments is a closure means that is broken or dislocated upon actuation (pressing, twisting, winding, rotating, bending, pushing, pulling, etc.). In another embodiment, the closure means comprises an integral removal means. In another embodiment, the closure means is a frangible seal.

[0055] In another embodiment, such elements are known to one of average skill in the art. In another embodiment, the compartments are rigid or flexible and may comprise various shapes such as a cylinder, a sack, a bag or an ampule. In another embodiment, at least one of the compartments housing a composition of the invention (in any form known to one of average skill in the art) comprises a sealing means that once removed provides an opening for dispensing the first composition, the second composition, or a mix comprising both the first and the second compositions. In another embodiment, the sealing means is a frangible seal.

[0056] In another embodiment, the kit houses each of the compositions described herein in a single use container. In another embodiment, the container comprises a molded deformable bag or bottle having a sealing means. In another embodiment, the kit of the invention comprises at least two components useful for of a mixture in distinct compartments.

[0057] In another embodiment, the kit includes two compartments wherein each compartment separately houses an active ingredient as described herein, wherein the active ingredients yield upon their admixture a composition for the relaxation of a smooth muscle.

In another embodiment, the kit includes two compartments wherein each compartment separately houses an active ingredient as described herein, wherein the active ingredients yield upon their admixture a composition for the relaxation of smooth muscles of Corpora Cavernosa.

[0058] In another embodiment, Isosorbide mono-nitrate, DMI or Isosorbide dinitrate is substituted or added to/with dimethyl Isosorbide. In another embodiment, a composition of the invention comprises dimethyl Isosorbide. In another embodiment, a composition of the invention comprising Isosorbide mono-nitrate, DMI or Isosorbide dinitrate further comprises dimethyl Isosorbide. In another embodiment, a composition of the invention comprises a mixture of the first and the second kit compositions.

[0059] In another embodiment, DMI is used as a skin penetration enhancer. In another embodiment, surprisingly, DMI has also a vasodilator activity within the composition of the invention.

[0060] In another embodiment, an effective amount is an amount that induces erection, treats ED, or treats female sexual arousal disorder in a subject. In another embodiment, an effective amount is an amount that induces erection, treats ED, or treats female sexual arousal disorder in a subject while minimizing side effects associated with the systemic administration of the APIs (as further discussed hereinbelow).

[0061] In another embodiment, FSAD is: lifelong (since birth) or acquired. In another embodiment, FSAD is based on context: occurs in all situations (generalized) or be situation- specific (situational).

[0062] In another embodiment, the phrase "Corpus Cavernosum" is Corpus Cavernosum penis and/or Corpus Cavernosum clitoridis.

[0063] In another embodiment, an active pharmaceutical ingredient (API) is a nitric oxide donor. In another embodiment, an API or the APIs is/are: L-Arginine, L-Norvaline,

Moxisylyte HC1, Hydralazine HC1, Isosorbide mono-nitrate, Isosorbide dinitrate, DMI or any combination thereof. In another embodiment, a composition of the invention comprises at least two APIs. In another embodiment, a composition of the invention comprises at least two APIs derived from the kit of the invention. In another embodiment, a composition of the invention comprises L-Arginine and L-Norvaline. In another embodiment, a composition of the invention comprises L-Arginine and Moxisylyte HCl. In another embodiment, a composition of the invention comprises L-Arginine and Hydralazine HCl. In another embodiment, a composition of the invention comprises L-Arginine and Isosorbide mono-nitrate, DMI or Isosorbide dinitrate. In another embodiment, a composition of the invention comprises Moxisylyte HCl and L-Norvaline. In another embodiment, a composition of the invention comprises Moxisylyte HCl and Hydralazine HCl. In another embodiment, a composition of the invention comprises Moxisylyte HCl and Isosorbide mono-nitrate, DMI or Isosorbide dinitrate. In another embodiment, a composition of the invention is the first composition, the second composition, or a mixture of the first and the second compositions.

[0064] In another embodiment, Isosorbide mono-nitrate, Isosorbide dinitrate, DMI, or any combination thereof have/has a vasodilator activity. In another embodiment, Isosorbide mono-nitrate, Isosorbide dinitrate, DMI, or any combination thereof have/has a penetration enhancing activity and vasodilator activity.

[0065] In another embodiment, a composition of the invention comprises L-Norvaline and Hydralazine HCl. In another embodiment, a composition of the invention comprises L- Norvaline and Isosorbide mono-nitrate, DMI or Isosorbide dinitrate. In another embodiment, a composition of the invention comprises: Hydralazine HCl, Moxisylyte HCl, Isosorbide mono-nitrate, DMI or Isosorbide dinitrate, Arginine, Norvaline or any combination thereof.

[0066] In another embodiment, a composition of the invention comprises: Hydralazine HCl, Moxisylyte HCl, L-Arginine and L-Norvaline. In another embodiment, a composition of the invention comprises: Hydralazine HCl, Isosorbide dinitrate (or DMI) and L-Arginine. In another embodiment, a composition of the invention comprises: Hydralazine HCl, Isosorbide dinitrate (or DMI), L-Arginine and L-Norvaline. In another embodiment, a composition of the invention comprises: Moxisylyte HCl, Isosorbide dinitrate, DMI or Isosorbide mono-nitrate and L-Arginine. In another embodiment, a composition of the invention comprises: Moxisylyte HCl, Isosorbide dinitrate, DMI or Isosorbide mono-nitrate, L-Arginine and L-Norvaline.

[0067] In another embodiment, the concentration of L-Arginine in a composition of the invention is between 2 to 100 μΜ. In another embodiment, the concentration of L-Arginine in a composition of the invention is between 5 to 50 μΜ. In another embodiment, the concentration of L-Arginine in a composition of the invention is between 5 to 30 μΜ. In another embodiment, the concentration of L-Arginine in a composition of the invention is between 15 to 30 μΜ. In another embodiment, the concentration of L-Arginine in a composition of the invention is between 5 to 20 μΜ. In another embodiment, the concentration of L-Arginine in a composition of the invention is between 1 to 20 μΜ. In another embodiment, the concentration of L-Arginine in a composition of the invention is between 5 to 15 μΜ.

[0068] In another embodiment, the concentration of L-Norvaline in a composition of the invention is between 5 to 100 μΜ. In another embodiment, the concentration of L-Norvaline in a composition of the invention is between 5 to 80 μΜ. In another embodiment, the concentration of L-Norvaline in a composition of the invention is between 20 to 100 μΜ. In another embodiment, the concentration of L-Norvaline in a composition of the invention is between 20 to 60 μΜ.

[0069] In another embodiment, the concentration of Moxisylyte HCl in a composition of the invention is between 0.01 to 50 μΜ. In another embodiment, the concentration of Moxisylyte HCl in a composition of the invention is between 0.1 to 10 μΜ. In another embodiment, the concentration of Moxisylyte HC1 in a composition of the invention is between 0.1 to 5 μΜ. In another embodiment, the concentration of Moxisylyte HC1 in a composition of the invention is between 0.01 to 0.5 μΜ. In another embodiment, the concentration of Moxisylyte HC1 in a composition of the invention is between 0.5 to 5 μΜ. In another embodiment, the concentration of Moxisylyte HC1 in a composition of the invention is between 0.5 to 2 μΜ.

[0070] In another embodiment, the concentration of Isosorbide mono-nitrate, DMI or Isosorbide dinitrate in a composition of the invention is between 0.001 to 1 μΜ. In another embodiment, the concentration of Isosorbide mono-nitrate, DMI or Isosorbide dinitrate in a composition of the invention is between 0.005 to 0.5 μΜ. In another embodiment, the concentration of Isosorbide mono-nitrate, DMI or Isosorbide dinitrate in a composition of the invention is between 0.005 to 0.1 μΜ. In another embodiment, the concentration of Isosorbide mono-nitrate, DMI or Isosorbide dinitrate in a composition of the invention is between 0.01 to 1 μΜ. In another embodiment, the concentration of Isosorbide mono-nitrate, DMI or Isosorbide dinitrate in a composition of the invention is between 0.01 to 0.5 μΜ.

[0071] In another embodiment, the concentration of Hydralazine HC1 in a composition of the invention is between 0.001 to 5 μΜ. In another embodiment, the concentration of Hydralazine HC1 in a composition of the invention is between 0.001 to 1 μΜ. In another embodiment, the concentration of Hydralazine HC1 in a composition of the invention is between 0.005 to 0.5 μΜ. In another embodiment, the concentration of Hydralazine HC1 in a composition of the invention is between 0.01 to 0.5 μΜ.

[0072] In another embodiment, the composition of the invention comprises L-Arginine and Moxisylyte HC1. In another embodiment, the composition of the invention comprises L- Arginine and Isosorbide dinitrate, DMI, or Isosorbide mono-nitrate. In another embodiment, the composition of the invention comprises Hydralazine HC1, DMI, Isosorbide dinitrate, Isosorbide mono-nitrate, Moxisylyte HC1, or any combination thereof. In another embodiment, the composition of the invention comprises Hydralazine HC1 and DMI, Isosorbide dinitrate, Isosorbide mono-nitrate.

[0073] As used herein, "a" or "an" means "at least one" or "one or more."

[0074] As used herein, an "erectile dysfunction" (or impotence) refers to the inability of a male mammal, e.g., a man, to achieve and maintain penile erection for satisfactory sexual intercourse.

[0075] As used herein, "female sexual arousal disorder" (FSAD) refers to the inability to attain sexual arousal or to maintain arousal until the completion of a sexual activity or to an inadequate lubrication-swelling response normally present during arousal and sexual activity.

[0076] As used herein, an "API" refers to a substance that increases sexual arousal stimulating activity.

[0077] As used herein, a "combination" refers to any association between two or among more items or APIs.

[0078] As used herein, a "composition" may refer to any mixture of two or more products or compounds, the first composition, or the second composition. It may be a solution, a suspension, liquid, powder, a paste, cream, gel, aerosol, aqueous, non-aqueous or any combination thereof.

[0079] As used herein, a "nerve dysfunction" refers, inter-alia to the inability of the penis to hold the blood during erection or the persistent or recurring inability to attain or maintain adequate sexual excitement causing personal distress including, but not limited to dysfunction caused by diabetes mellitus, hypertension, hyperlipidemia, penile injury, aging, pelvic surgery or irradiation. [0080] As used herein, an "arterial insufficiency" refers to reduced perfusion pressure and arterial flow associated with trauma or disease including, but not limited to, that associated with hypertension, hyperlipidemia, cigarette smoking, diabetes mellitus, and pelvic irradiation.

[0081] As used herein, a "venous leakage" refers to the inability of the penis to hold the blood during erection including, but not limited to diabetes mellitus, hypertension, hyperlipidemia, penile injury, aging, pelvic surgery or irradiation.

[0082] As used herein, a "drug use" includes pharmaceutical drug use and substance abuse. Permeation/penetration enhancer

[0083] In another embodiment, a composition of the invention comprises a topical carrier having superior skin penetration capability. In another embodiment, a topical carrier having superior skin penetration capability is a permeation/penetration enhancer which increases the penetration of the APIs into or through the skin to the target organelle. In another embodiment, a topical carrier comprises petrolatum, a bodying agent such as cetyl alcohol, stearyl alcohol, cetearyl alcohol (a mixture of cetyl alcohol and stearyl alcohol) or stearic acid, and a solubility enhancer such as propylene glycol, hexylene glycol, polyethylene glycol or transcutol as a penetration enhancer.

[0084] In another embodiment, a topical carrier comprises methyl laurate, oleic acid, myristyl alcohol, or any combination thereof. In another embodiment, a topical carrier comprises N-(2-hydroxyethyl)pyrrolidone. In another embodiment, a topical carrier comprises oleic acid, oleyl alcohol or a combination thereof. In another embodiment, a topical carrier comprises lauryl alcohol (1-dodecanol). In another embodiment, a topical carrier comprises a monoalcohol. In another embodiment, a topical carrier comprises a pyrrolidone-type solvent. In another embodiment, a topical carrier comprises a penetration/permeation enhancer selected from the group comprising of: benzyl alcohol, propylene glycol, ethoxydiglycol, transcutol or any combination thereof. In another embodiment, a topical carrier comprises alkyl-2-(N-substituted amino)-alkanoate, an (N- substituted amino)-alkanol alkanoate, or a mixture of these. In another embodiment, a topical carrier comprises at least one penetration/permeation enhancer. In another embodiment, the penetration/permeation enhancer is DDAIP, an ester of Ν,Ν-dimethylalanine and dodecanol. In another embodiment, DDAIP is formulated in its hydrochloride salt (DDAIP.HCl).

[0085] In another embodiment, the penetration/permeation enhancer is a nanoparticle encapsulating or carrying the APIs of the invention. In another embodiment, the penetration/permeation enhancer is a liposome encapsulating the APIs of the invention.

[0086] In another embodiment, a topical carrier comprises a penetration/permeation enhancer comprising aery late polymers such as: acrylic acid polymer, methacrylic acid polymer, alkyl acrylate polymer, alkyl methacrylate polymer or copolymers which effectively control skin penetration of the APIs. In another embodiment, a topical carrier is an alcoholic or aqueous alcoholic gel comprising 2-n-nonyl-l,3-dioxolane or other hydrocarbyl derivative of 1,3-dioxolane-or 1,3-dioxane or acetal. In another embodiment, a topical carrier is a fatty acid ester. In another embodiment, the fatty acid ester is: glycerol monolaurate, glycerol monooleate, glycerol monolinoleate, glycerol trilaurate, glycerol trioleate, glycerol tricaprylate, propylene glycol monolaurate, propylene glycol dilaurate, caprylic/capric triglyceride, methyl laurate, methyl caprate, isopropyl myristate, isopropyl palmitate, ethyl oleate and oleyl oleate. In another embodiment, a topical carrier comprises an alkyl lactate such as but not limited to: ethyl lactate.

[0087] In another embodiment, a topical carrier is a carrier capable of carrying the APIs of the invention to the Corpus Cavernosum. In another embodiment, a topical carrieris a carrier capable of carrying the APIs of the invention to the Tunica Albugines. In another embodiment, a topical carrier is a carrier capable of carrying the APIs of the invention to the Cavernosa! Space. In another embodiment, a topical carrier is a carrier capable of carrying the APIs of the invention to the Cavernosal Artery. In another embodiment, a topical carrier is a carrier capable of carrying the APIs of the invention to the terminations of the cavernous nerve, to the endothelium lining of arterial distributions, to the sinusoidal spaces, or any combination thereof.

Nitric oxide donor

[0088] In another embodiment, the nitric oxide donor is a small molecule. In another embodiment, the nitric oxide donor affects the L-Arginine/NO/cyclic guano sine monophosphate (cGMP)/cyclic adenosine monophosphate (cAMP) pathways.

[0089] In another embodiment, the nitric oxide donor is Hydralazine or a salt thereof. In another embodiment, the nitric oxide donor is Arginine. In another embodiment, the nitric oxide donor is Moxisylyte or a salt thereof. In another embodiment, Alprostadil can

substitute any of the active ingredients described herein ^{n0 'J} ' . In

another embodiment, the nitric oxide donor is L- Arginine . In another embodiment, the nitric oxide donor is DMI, Isosorbide dinitrate, Isosorbide mononitrate. In another embodiment, the nitric oxide donor is Hydralazine hydrochloride

. In another embodiment, the nitric oxide donor is nitroglycerin. Additional potential APIs [0090] In one embodiment, an API of the invention is Milrinone

another embodiment, an API of the invention is Forskolin . In another

embodiment, an API of the invention is Prazosin . In another embodiment, Milrinone, Forskolin or both is/are combined with any API described herein.

Causes for ED or female sexual arousal disorder

[0091] In another embodiment, ED is a consequence of inadequate penile blood supply. In another embodiment, ED is a consequence of psychological problems. In another embodiment, ED is a consequence of a chronic medical illness. In another embodiment, ED is a consequence of effect of certain drugs.

[0092] In another embodiment, the compositions and/or kits of the invention are used for treating a subject afflicted with ED (male) or female sexual arousal disorder. In another embodiment, the compositions of the invention are used topically for treating a subject afflicted with ED (male) or female sexual arousal disorder. In another embodiment, the compositions and/or kits of the invention are used for treating a subject afflicted with ED or female sexual arousal disorder regardless of the underlying cause as long as the erection mechanisms in the subject are potentially functional. In another embodiment, the cause for ED or female sexual arousal disorder is unknown. In another embodiment, the cause for ED or female sexual arousal disorder (FSAD) is a vascular disease. In another embodiment, the cause for ED or FSAD is Diabetes mellitus. In another embodiment, the cause for ED is hypogonadism. In another embodiment, the cause for ED or FSAD is hyperprolactinemia. In another embodiment, the cause for ED or FSAD is hypothyroidism. In another embodiment, the cause for ED or FSAD is hyperthyroidism. In another embodiment, the cause for ED or FSAD is Cushing's syndrome. In another embodiment, the cause for ED or FSAD is alcoholism. In another embodiment, the cause for ED or FSAD is drug abuse. In another embodiment, the cause for ED or FSAD is neurologic. In another embodiment, the cause for ED or FSAD is psychogenic. In another embodiment, the cause for ED or FSAD is pharmacologic.

[0093] In another embodiment, the cause for ED or FSAD is use of an antihypertensive drug. In another embodiment, the cause for ED or FSAD is use of a beta blocker. In another embodiment, the cause for ED or FSAD is use of a diuretic. In another embodiment, the cause for ED or FSAD is use of an antidepressant. In another embodiment, the cause for ED or FSAD is use of an antipsychotic drug. In another embodiment, the cause for ED or FSAD is use of spironolactone. In another embodiment, the cause for ED or FSAD is use of cimetidine. In another embodiment, the cause for ED or FSAD is use of finasteride. In another embodiment, the cause for ED or FSAD is use of a monoamine oxidase inhibitor. In another embodiment, the cause for ED or FSAD is use of a selective serotonin reuptake inhibitor. In another embodiment, the cause for ED or FSAD is use of a tricyclic antidepressant. In another embodiment, the cause for ED or FSAD is use of clonidine. In another embodiment, the cause for ED or FSAD is use of methyldopa. In another embodiment, the cause for ED or FSAD is use of an anxiolytics agent. In another embodiment, the cause for ED or FSAD is use of a corticosteroid. In another embodiment, the cause for ED or FSAD is use of gemfibrozil. In another embodiment, the cause for ED or FSAD is use of an anabolic steroid. [0094] In another embodiment, the cause for ED or FSAD is use of heroin. In another embodiment, the cause for ED or FSAD is use of marijuana. In another embodiment, the cause for ED or FSAD is use of a gonadotropin-releasing hormone agonist. In another embodiment, the cause for ED or FSAD is use of an antiandrogen, antiestrogen, and/or hormonal therapy. In another embodiment, the cause for FSAD is use of an estrogen- containing medication. In another embodiment, the cause for ED or FSAD is use of disopyramide. In another embodiment, the cause for ED or FSAD is use of digoxin.

Advantages over the known medications

[0095] In another embodiment, the compositions and/or kits of the invention are free of the devastating side effects associated with ED or FSAD, oral or injectable, medications such as but not limited to: headache, hypotension, chest pain, flushing, dyspepsia, rhinitis, or change in perception of color. In another embodiment, the compositions and/or kits of the invention enable intercourse to patients suffering from cardiovascular diseases.

[0096] In another embodiment, the methods of the present invention reduce side effects associated with systemic or oral ED or FSAD medications such as: dizziness, anxiety, nausea, a severe drop in blood pressure, abdominal pain, fatigue, hallucinations, paralysis, polymorphic ventricular tachycardia (fast heart rhythm), arterial hypotension, hyperhidrosis, constipation, vertigo, increased transaminase levels, increased alkaline phosphatase levels, somnolence (strong desire for sleep), and interference with sulphobromophthalein (dye used to determine hepatic function), paradoxical aggravation of cerebral vasospasm, headache, facial flushing, nasal congestion, upset stomach, loss of appetite, jaundice, eosinophilia, thrombopenia, mixed hepatitis, abnormal vision as well as isolated reports of hearing and vision loss. In another embodiment, the methods of the present invention overcome the devastating risks and discomforts associated with Intra-penile injections or intra-corporeal injections. Use of the composition and/or kit

[0097] In another embodiment, a composition of the invention or a mixture of the first and the second compositions derived from the kit is applied over the penis, glans penis, or the urethral opening 2 to 60 minutes before intercourse. In another embodiment, a composition of the invention or a mixture of the first and the second compositions derived from the kit is applied over the glans penis 2 to 30 minutes before intercourse. In another embodiment, a composition of the invention or a mixture of the first and the second compositions derived from the kit is applied over the glans penis 5 to 25 minutes before intercourse. In another embodiment, a composition of the invention or a mixture of the first and the second compositions derived from the kit is applied over the glans penis 10-30 minutes before intercourse. In another embodiment, a composition of the invention or a mixture of the first and the second compositions derived from the kit is applied over the glans penis 15 to 40 minutes before intercourse. In another embodiment, a composition of the invention or a mixture of the first and the second compositions derived from the kit is applied over the glans penis 15 to 25 minutes before intercourse.

Methods

[0098] In one embodiment, the present invention provides a method for preventing or treating male erectile dysfunction (ED) or female sexual arousal disorder (FSAD), which method comprises administering to a mammal to whom such prevention or treatment is needed or desirable, an effective amount of a composition or a mixture of the first and the second compositions derived from the kit, as described herein. In another embodiment, male ED or female FSAD in any mammal can be treated or prevented by the present method. Preferably, male ED or female FSAD in humans is treated or prevented.

[0099] In another embodiment, the present invention provides a method for the treatment or prevention of male ED induced by or secondary to nerve dysfunction, arterial insufficiency, venous leakage, hormonal insufficiency, drug use, surgery, chemotherapy or radiation. In another embodiment, the present invention provides for a combination of APIs of the invention, for preventing or treating male ED or female FSAD.

[00100] In another embodiment, the magnitude of a therapeutic dose in the acute or chronic treatment of male ED or female FSAD will vary with the severity of the condition to be treated and the route of administration. The dose, and perhaps dose frequency, will also vary according to age, body weight, condition and response of the individual patient as can be assessed by a physician.

[00101] In one embodiment, the present invention contemplates the administration by the application of a topical formulation or a combination derived from a kit, such as but not limited to a cream or a gel in an amount sufficient to improve blood flow, blood volume, and regenerate nerve and smooth muscle function in the penis.

[00102] In another embodiment, the present invention provides a method of treating a male suffering from ED or a female suffering from FSAD, comprising the step of administering to the male or the female a composition as described herein. In another embodiment, the present invention provides a method of treating a male suffering from ED or a female suffering from FSAD, comprising the step of administering to the male or the female a composition comprising an effective amount of: Isosorbide mono-nitrate, DMI or Isosorbide dinitrate, a nitric oxide donor, and a pharmaceutical acceptable carrier, thereby treating a male suffering from ED or a female suffering from FSAD. In another embodiment, erectile dysfunction is the inability to develop or maintain an erection of the penis during sexual performance. In another embodiment, erectile dysfunction is a difficulty in producing erection. In another embodiment, the method of the invention provides a first line treatment of ED or FSAD. [00103] In another embodiment, "treating" is ameliorating or improving ED/FSAD, orgasmic function, sexual desire, intercourse satisfaction, overall satisfaction, or any combination thereof.

[00104] In another embodiment, the methods of the invention induce a reflex erection. In another embodiment, the methods of the invention induce a psychogenic erection. In another embodiment, the methods of the invention induce the relaxation of smooth muscles of Corpora Cavernosa.

[00105] In another embodiment, the phrase "inducing erection" comprises inducing de- novo erection, prolonging an erection, maintaining an erection, increasing blood flow or blood volume in the Corpus Cavernosum. In another embodiment, the phrase "inducing erection" is increasing penile rigidity. In another embodiment, the phrase "inducing erection" comprises the induction of reflex erection. In another embodiment, the phrase "inducing erection" comprises the induction of psychogenic erection. In another embodiment, the phrase "inducing erection" comprises the induction of relaxation of smooth muscles in the Corpora Cavernosa. In another embodiment, the phrase "inducing erection" comprises inducing neurogenic NO. In another embodiment, the phrase "inducing erection" comprises activating soluble guanylyl cyclase. In another embodiment, the phrase "inducing erection" comprises increasing the NO level in the corpus cavernosum. In another embodiment, the phrase "inducing erection" comprises increasing NO production in the endothelium and/or the nerves innervating the corpus cavernosum.

[00106] In one embodiment, the subject is a male. In another embodiment, the subject is a female. In another embodiment, the subject is a male afflicted with ED or a female afflicted with FSAD. In another embodiment, the subject is a subject afflicted with a circulatory disease. In another embodiment, the subject is a subject afflicted with a cardiovascular disease. In another embodiment, the subject is a subject afflicted with diabetes. In another embodiment, the subject is a subject afflicted with a neurological pathology. In another embodiment, the subject is a subject afflicted with hormonal imbalance. In another embodiment, the subject is a subject afflicted with drug side effects. In another embodiment, the subject is a subject afflicted with psychological impotence. In another embodiment, the subject is a subject afflicted with kidney failure. In another embodiment, the subject is a subject afflicted with multiple sclerosis. In another embodiment, the subject is a heavy smoker. In another embodiment, the subject is a subject that underwent prostate and/or bladder surgery. In another embodiment, the subject is a subject afflicted with a Cavernosal disorder.

[00107] In another embodiment, the present invention provides a method for inducing a penile erection in a subject, comprising the step of administering to the subject a composition comprising an effective amount of: Isosorbide mono-nitrate, DMI or Isosorbide dinitrate, a nitric oxide donor, Moxisylyte, Hydralazine, or any combination thereof and a pharmaceutical acceptable carrier, thereby inducing erection in a subject. In another embodiment, the present invention provides a method for maximizing a penile erection in a subject, comprising the step of administering to the subject a composition comprising an effective amount of: Isosorbide mono-nitrate or Isosorbide dinitrate, DMI, Moxisylyte, Hydralazine, or any combination thereof, and a pharmaceutical acceptable carrier, thereby maximizing erection in a subject. In another embodiment, the present invention provides a method for prolonging a penile erection in a subject, comprising the step of administering to the subject a composition comprising an effective amount of: Isosorbide mono-nitrate or Isosorbide dinitrate, a nitric oxide donor, and a pharmaceutical acceptable carrier, thereby prolonging an erection in a subject.

[00108] In another embodiment, the subject does not suffer from ED but rather desires to prolong an erection. In another embodiment, the subject does not suffer from ED but rather desires to induce bigger erection, harder erection, longer erection, or any combination thereof.

[00109] In another embodiment, the present invention provides a method for inducing neurogenic NO for relaxation of penile vessels and Corpus Cavernosum. In another embodiment, the present invention provides a method for inducing smooth muscle relaxation in the penis. In another embodiment, the present invention provides a method for inducing penile nitric oxide (NO) thus activating penile soluble guanylyl cyclase. In another embodiment, the present invention provides a method for increasing the NO level in the corpus cavernosum. In another embodiment, the present invention provides a method for increasing NO production in the endothelium and/or the nerves innervating the corpus cavernosum.

[00110] In another embodiment, administering is applying topically to the penis. In another embodiment, the methods of the invention induce/increase localized blood flow in penile tissue (Corpus Cavernosum), the method comprising the step of: applying topically to the skin a composition of the invention.

Increasing the erectile effect of a PDE-5 inhibitor

[00111] In one embodiment, provided herein a method for increasing and/or enhancing the erectile induction effect of a PDE-5 inhibitor in a subject in need thereof, comprising administering to the subject a composition comprising: (a) Isosorbide mono-nitrate, DMI, or Isosorbide dinitrate; (b) a Hydralazine salt, Hydralazine, Arginine, Moxisylyte, or a

Moxisylyte salt; or a combination of (a) and (b), thereby increasing and/or enhancing the erectile effect of a PDE-5 inhibitor in a subject. In one embodiment, the composition comprising: (a) Isosorbide mono-nitrate, DMI, or Isosorbide dinitrate; (b) a Hydralazine salt,

Hydralazine, Arginine, Moxisylyte, or a Moxisylyte salt; or a combination of (a) and (b), is administered prior to the administration of a PDE-5 inhibitor or is administered concomitantly with the PDE-5 inhibitor. In one embodiment, the PDE-5 inhibitor is Viagra. In one embodiment, provided herein a method for increasing and/or enhancing the erectile induction comprising combining a PDE-5 inhibitor and a composition as described herein in a single composition or as separate compositions in a single treatment.

[00112] In one embodiment, provided herein a method for rendering a refractory or ineffective clinical dose of a PDE-5 inhibitor, in a subject, effective, comprising administering to the subject a composition comprising: (a) Isosorbide mono-nitrate, DMI, or Isosorbide dinitrate; (b) a Hydralazine salt, Hydralazine, Arginine, Moxisylyte, or a Moxisylyte salt; or a combination of (a) and (b), thereby rendering a refractory or ineffective clinical dose of a PDE-5 inhibitor -effective. In another embodiment, a subject as described herein has no ED improvement after treatment with a dose providing cure to other ED patients that are responsive to the PDE-5 inhibitor. In another embodiment, a subject as described herein is non-responsive to a PDE-5 inhibitor within the known recommended dose range. In another embodiment, a subject as described herein is non-responsive to a PDE-5 inhibitor, therefore an otherwise effective dose is refractory or ineffective clinical dose of a PDE-5 inhibitor in this non-responsive subject. In one embodiment, a composition as described herein renders a refractory or an ineffective clinical dose of a PDE-5 inhibitor- effective.

[00113] In one embodiment, provided herein a composition comprising a PDE-5 inhibitor and Isosorbide mono-nitrate, DMI, or Isosorbide dinitrate. In one embodiment, provided herein a composition comprising a PDE-5 inhibitor and a Hydralazine salt, Hydralazine, Arginine, Moxisylyte, or a Moxisylyte salt. In one embodiment, provided herein a composition comprising: (a) a PDE-5 inhibitor, (b) a Hydralazine salt, Hydralazine, Arginine, Moxisylyte, or a Moxisylyte salt, and (c) Isosorbide mono-nitrate, DMI, or Isosorbide dinitrate. [00114] In another embodiment, a refractory or ineffective clinical dose of a PDE-5 inhibitor according to the present invention is the dose providing cure to ED patients that are responsive to the PDE-5 inhibitor. In one embodiment, a subject as described herein is non- responsive (with respect to erectile induction or enhancement) to the recommended dose range of the PDE-5 inhibitor. In another embodiment, a refractory or ineffective clinical dose of a PDE-5 inhibitor is the quantity of a PDE-5 inhibitor known to be required to cure ED and/or symptoms thereof, in a subject suffering from ED (according to medical literature such as but not limited to PDR). Thus, the invention provides means for using a PDE-5 inhibitor within the therapeutic recommended range of the PDE-5 inhibitor, in subjects that were not responsive to the therapeutic recommended range of the PDE-5 inhibitor. In another embodiment, a refractory or ineffective clinical dose of a PDE-5 inhibitor is the dose providing cure to patients that are responsive to PDE-5 inhibitor with respect to correcting ED manifestations. In another embodiment, a refractory or ineffective clinical dose of Viagra, as described herein, is 25 to 100 mg.

Diagnosis of ED or FS AD

[00115] In another embodiment, during the diagnosis process of ED or FSAD some blood tests are done to exclude an underlying disease, such as hypogonadism and prolactinoma. In another embodiment, diagnosis includes the use of the guidelines provided in the current diagnostic and statistical manual of mental diseases (DSM-rV). In another embodiment, diagnosis includes the use of duplex ultrasound to evaluate blood flow, venous leak, signs of atherosclerosis, and scarring or calcification of erectile tissue. In another embodiment, diagnosis includes the injection of prostaglandin for inducing erection, measuring penile blood pressure, and viewing vascular dilation. In another embodiment, diagnosis includes the bulbocavernosus reflex test are which evaluates nerve sensation in the penis. [00116] In another embodiment, a physician squeezes the glans (head) of the penis, which immediately causes the anus to contract if nerve function is normal. In another embodiment, a physician measures the latency between squeeze and contraction by observing the anal sphincter or by feeling it with a gloved finger inserted past the anus.

[00117] In another embodiment, diagnosis includes the Nocturnal penile tumescence (NPT). In another embodiment, changes in penile rigidity are measured during nocturnal erection: snap gauge and strain gauge.

[00118] In another embodiment, diagnosis includes a penile biothesiometry test. In another embodiment, a penile biothesiometry test utilizes electromagnetic vibration to evaluate sensitivity and nerve function in the glans and shaft of the penis.

[00119] In another embodiment, diagnosis includes the dynamic infusion Cavemosometry (DICC) test. In another embodiment, DICC test includes the pumping of fluid into the penis at a known rate and pressure, to evaluate the vascular pressure in the corpus cavemosum during an erection.

[00120] In another embodiment, diagnosis includes the Corpus Cavemosometry test. In another embodiment, the Corpus Cavemosometry test measures the vascular pressure in the Corpus Cavemosum: saline is infused under pressure into the Corpus Cavemosum with a butterfly needle, and the flow rate needed to maintain an erection indicates the degree of venous leakage.

[00121] In another embodiment, diagnosis includes Magnetic resonance angiography (MRA) test. Blood supply and vascular anomalies are assessed by injecting a "contrast agent" into the subject's bloodstream that causes vascular tissues to stand out against other tissues.

Amounts/ratios of APIs [00122] In another embodiment, an API as described herein is generally employed in therapeutic amounts as indicated in the PHYSICIANS' DESK REFERENCE (PDR) (2012), which is incorporated herein by reference, or such therapeutically useful amounts as would be known to one of ordinary skill in the art. The compounds of the invention and any other ingredient can be administered at the recommended maximum clinical dosage or at lower doses. Dosage levels of the active compounds in the compositions of the invention may be varied to obtain a desired therapeutic response depending on the route of administration, severity of the disease and the response of the patient. The combination can be administered as separate compositions or as a single dosage form containing both agents. When administered as a combination (such as a single composition or a mixture derived from a kit), the therapeutic agents can be formulated as separate compositions that are given at the same time or different times, or the therapeutic agents can be given as a single composition.

[00123] In another embodiment, the molar concentration of Moxisylyte or a salt thereof is at least 2 folds greater than the molar concentration of the Isosorbide mono-nitrate or Isosorbide dinitrate. In another embodiment, the molar concentration of Moxisylyte or a salt thereof is at least 3 folds greater than the molar concentration of the Isosorbide mono-nitrate or Isosorbide dinitrate. In another embodiment, the molar concentration of Moxisylyte or a salt thereof is at least 2-5 folds greater than the molar concentration of the Isosorbide mononitrate or Isosorbide dinitrate. In another embodiment, the molar concentration of Moxisylyte or a salt thereof is at least 3-10 folds greater than the molar concentration of the Isosorbide mono-nitrate or Isosorbide dinitrate. In another embodiment, the molar concentration of Moxisylyte or a salt thereof is at least 3-6 folds greater than the molar concentration of the Isosorbide mono-nitrate or Isosorbide dinitrate.

[00124] In another embodiment, Hydralazine or a salt thereof is 0.01 to 5 μΜ Hydralazine or a salt thereof. In another embodiment, Hydralazine or a salt thereof is 0.05 to 2 μΜ Hydralazine or a salt thereof. In another embodiment, Hydralazine or a salt thereof is 0.05 to 1 μΜ Hydralazine or a salt thereof. In another embodiment, Hydralazine or a salt thereof is 0.1 to 0.5 μΜ Hydralazine or a salt thereof.

[00125] In another embodiment, Isosorbide mono-nitrate, DMI or Isosorbide dinitrate is 0.0005 to 5 μΜ Isosorbide mono-nitrate or Isosorbide dinitrate. In another embodiment, Isosorbide mono-nitrate, DMI or Isosorbide dinitrate is 0.001 to 1 μΜ Isosorbide mononitrate or Isosorbide dinitrate. In another embodiment, Isosorbide mono-nitrate, DMI or Isosorbide dinitrate is 0.005 to 0.5 μΜ Isosorbide mono-nitrate or Isosorbide dinitrate. In another embodiment, Isosorbide mono-nitrate, DMI or Isosorbide dinitrate is 0.01 to 0.1 μΜ Isosorbide mono-nitrate or Isosorbide dinitrate.

[00126] In another embodiment, Arginine is 0.1 to 100 μΜ Arginine. In another embodiment, Arginine is 0.5 to 50 μΜ Arginine. In another embodiment, Arginine is 1 to 50 μΜ Arginine. In another embodiment, Arginine is 5 to 50 μΜ Arginine. In another embodiment, Arginine is 0.5 to 10 μΜ Arginine. In another embodiment, Arginine is L- Arginine.

[00127] In another embodiment, Moxisylyte or a salt thereof is 0.0001 to 10 μΜ Moxisylyte or a salt thereof. In another embodiment, Moxisylyte or a salt thereof is 0.0005 to 5 μΜ Moxisylyte or a salt thereof. In another embodiment, Moxisylyte or a salt thereof is 0.001 to 1 μΜ Moxisylyte or a salt thereof. In another embodiment, Moxisylyte or a salt thereof is 0.005 to 0.5 μΜ Moxisylyte or a salt thereof. In another embodiment, Moxisylyte or a salt thereof is 0.005 to 0.1 μΜ Moxisylyte or a salt thereof. In another embodiment, Moxisylyte or a salt thereof is 0.01 to 0.5 μΜ Moxisylyte or a salt thereof.

[00128] In one embodiment, the present invention provides combined preparations . In one embodiment, the term "a combined preparation" defines especially a "kit of parts" in the sense that the combination partners as defined above can be dosed independently or by use of different fixed combinations with distinguished amounts of the combination partners i.e., simultaneously, concurrently, separately or sequentially. In some embodiments, the parts of the kit of parts can then, e.g., be administered simultaneously or chronologically staggered, that is at different time points and with equal or different time intervals for any part of the kit of parts. In some embodiments, the parts of the kit of parts are mixed within a closed compartment within the kit ad are then (in a mixed form) applied topically onto the skin. In some embodiments, applied topically onto the skin involves mixing of the parts. In another embodiment the parts are the first composition and the second composition as described herein. The ratio of the total amounts of the combination partners, in some embodiments, can be administered in the combined preparation. In one embodiment, the combined preparation can be varied, e.g., in order to cope with the needs of a patient subpopulation to be treated or the needs of the single patient which different needs can be due to a particular disease, severity of a disease, age, sex, or body weight as can be readily made by a person skilled in the art.

The pharmaceutical compositions

[00129] The pharmaceutical compositions comprising an API or the APIs as described herein are administered to a subject by any method known to a person skilled in the art, such as topically, parenterally, paracancerally, transmucosally, transdermally, intramuscularly, intravenously, intradermally, subcutaneously, intraperitonealy, intraventricularly, intracranially, intravaginally or intratumorally.

[00130] In another embodiment, a variety of means for effecting or improving absorption of the APIs are envisioned. In another embodiment, the APIs are packaged in such a way that it is carried into a target tissue in a male or a female (Corpus Cavernosum) by derivitization or forming a neutral salt. [00131] In another embodiment, an API or the APIs are packaged liposomes or emulsions of collagen, collagen peptides or other components of skin or basement membrane. In another embodiment, an API or the APIs are delivered in a composition which is effective in neutralization of charge including delivery of the APIs in the form or an ester or salt which is electronically neutral.

[00132] In another embodiment, absorption is further aided by combining the use of hostile biophysical environments with the use of penetrating agents such as but not limited to: oleoresin capsicum or its constituents or molecules containing heterocyclic rings to which are attached hydrocarbon chains.

[00133] The compositions of the present invention can optionally include as active ingredients nettles and/or zinc. The components are widely available commercially.

[00134] In another embodiment, the compositions of the present invention include other ingredients that are not physiologically active but serve to enhance the pharmaceutical elegance of the final topical composition. For example, the compositions of the present invention may include excipients such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidinone, cellulose, water, syrup, and methyl cellulose. In some embodiments, the compositions of the present invention include lubricating agents such as talc, magnesium stearate and mineral oil, wetting agents, emulsifying and suspending agents, preserving agents such as methyl- and propylhydroxybenzoates, sweetening agents or flavoring agents.

[00135] In another embodiment, a composition as described herein comprises ISDN (or

ISDN/Lactose), Moxisylyte, ethanol, water and one or more of the group consisting of:

Methyl paraben, Transcutol P, PEG 400, and Propyl paraben. In another embodiment, a composition as described herein comprises ISDN (or ISDN/Lactose), Moxisylyte, ethanol, water, and PEG 400. In another embodiment, a composition as described herein comprises ISDN (or ISDN/Lactose), Moxisylyte, ethanol, water and at least one of Methyl paraben and Propyl paraben. In another embodiment, a composition as described herein comprises ISDN (or ISDN/Lactose), Moxisylyte, ethanol, water, Methyl paraben, Transcutol P, PEG 400, and Propyl paraben.

[00136] In another embodiment, the compositions of the present invention may be formulated in any pharmaceutically acceptable topical vehicle that does not interact adversely with the active ingredients. In another embodiment, compositions of the present invention may be formulated in water or oil based topical vehicles. These compositions, in some embodiments, can include lanolin, aquaphor, methylcellulose and derivatives thereof, petroleum based vehicles, aloe and the like. In another embodiment, the compositions of the present invention are formulated in a topical, water-based vehicle containing aloe and vitamin E.

[00137] In another embodiment, the topical compositions of the present invention include a mixture of a powder, distilled water oil, stearic acid, an alcohol, an emulsifying wax, glycerin, palmitic acid, denatured alcohol, methyl salicylate, lecithin, sodium bicarbonate, ascorbyl palmitate, polysorbate, methylparaben, propylparaben, or any combination thereof. In another embodiment, the topical compositions of the present invention have a pH of between about 5.0 and about 8.

[00138] In another embodiment, the topical compositions or combinations of the present invention are formulated for transdermal delivery of the active ingredients following topical administration to the penis or labia majora and minora pudendi. In another embodiment, a small amount of the compositions of the present invention is applied topically directly to the penis or labia majora and minora pudendi. In another embodiment, the topical compositions of the present invention are packaged as a coating on the interior surface of a condom, thereby, upon use of the condom, the composition is applied topically to the penis. [00139] If the preferred mode the combined formulation is administered orally, in one embodiment, a unit dosage form used may comprise tablets, capsules, lozenges, chewable tablets, suspensions, emulsions and the like. Such unit dosage forms comprise a safe and effective amount of the desired compound, or compounds, each of which is in one embodiment, from about 0.1 or 500 mg of each API described herein, or in another embodiment, about 0.5 or 10 mg. Pharmaceutically-acceptable carriers suitable for the preparation of unit dosage forms for peroral administration are well-known in the art. Tablets typically comprise conventional pharmaceutically-compatible adjuvants as inert diluents, such as calcium carbonate, sodium carbonate, mannitol, lactose and cellulose; binders such as starch, gelatin and sucrose; disintegrants such as starch, alginic acid and croscarmelose; lubricants such as magnesium stearate, stearic acid and talc. Glidants such as silicon dioxide can be used to improve flow characteristics of the powder-mixture. Coloring agents, such as the FD&C dyes, can be added for appearance. Sweeteners and flavoring agents, such as aspartame, saccharin, menthol, peppermint, and fruit flavors, are useful adjuvants for chewable tablets. Capsules typically comprise one or more solid diluents disclosed above. The selection of carrier components depends on secondary considerations like taste, cost, and shelf stability, which are not critical for the purposes of this invention, and can be readily made by a person skilled in the art.

[00140] In one embodiment, the oral dosage form comprises predefined release profile. In one embodiment, the oral dosage form of the present invention comprises an extended release tablets, capsules, lozenges or chewable tablets. In one embodiment, the oral dosage form of the present invention comprises a slow release tablets, capsules, lozenges or chewable tablets. In one embodiment, the oral dosage form of the present invention comprises an immediate release tablets, capsules, lozenges or chewable tablets. In one embodiment, the oral dosage form is formulated according to the desired release profile of the pharmaceutical active ingredient as known to one skilled in the art.

[00141] Peroral compositions may comprise liquid solutions, emulsions, suspensions, and the like. The pharmaceutically-acceptable carriers suitable for preparation of such compositions are well known in the art. Such liquid oral compositions comprise, in some embodiments, from about 0.001% to about 0.933% of each of the APIs, or in another embodiment, from about 0.033% to about 0.7%.

[00142] Compositions for use in the methods of this invention may comprise powders, solutions or emulsions, which in some embodiments are aqueous solutions or emulsions comprising a safe and effective amount of the APIs and optionally, other compounds, intended for topical intranasal administration. Such compositions may comprise from about 0.01% to about 10.0% w/v of any subject compound, more preferably from about 0.1% to about 2.0, which may be used for systemic delivery of the compounds by the intranasal route.

[00143] Other compositions and kits comprise dry powders of any one or more of the active ingredients described herein. Other compositions may be formulated for atomization and/or inhalation administration. Such compositions may be contained in a container with attached atomizing means.

[00144] Further, in another embodiment, the pharmaceutical compositions are administered by intravenous, intra-arterial, or intramuscular injection of a liquid preparation. Suitable liquid formulations include solutions, suspensions, dispersions, emulsions, oils and the like. In one embodiment, the pharmaceutical compositions are administered intravenously, and are thus formulated in a form suitable for intravenous administration. In another embodiment, the pharmaceutical compositions are administered intra-arterially, and are thus formulated in a form suitable for intra-arterial administration. In another embodiment, the pharmaceutical compositions are a mixed formulation of the kit's ingredients. In another embodiment, the pharmaceutical compositions are administered intramuscularly, and are thus formulated in a form suitable for intramuscular administration.

[00145] Further, in another embodiment, the pharmaceutical compositions including a mixture of the kit's compositions are administered topically to body surfaces, and are thus formulated in a form suitable for topical administration. Suitable topical formulations include gels, ointments, creams, lotions, drops and the like. For topical administration, the APIs or their physiologically tolerated derivatives such as salts, esters, N-oxides, and the like are combined with an additional appropriate therapeutic agent or agents, prepared and applied as solutions, suspensions, or emulsions in a physiologically acceptable diluent with a pharmaceutical carrier.

[00146] In another embodiment, the active compounds described herein, can be delivered in a vesicle, in particular a liposome (see Langer, Science 249:1527-1533 (1990); Treat et al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez- Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez -Berestein, ibid., pp. 317-327; see generally ibid).

[00147] In another embodiment, the pharmaceutical composition delivered in a controlled release system, transdermal patch, liposomes, or other modes of administration. In one embodiment, a pump may be used (see Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng. 14:201 (1987); Buchwald et al., Surgery 88:507 (1980); Saudek et al., N. Engl. J. Med. 321:574 (1989). In another embodiment, polymeric materials can be used. In yet another embodiment, a controlled release system can be placed in proximity to the therapeutic target, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138 (1984). Other controlled release systems are discussed in the review by Langer (Science 249:1527-1533 (1990). [00148] Numerous additional administration vehicles will be apparent to those of ordinary skill in the art, including without limitation slow release formulations, liposomal formulations and polymeric matrices.

[00149] The preparation of pharmaceutical compositions which contain active components is well understood in the art, for example by mixing, granulating, or tablet- forming processes. The active therapeutic ingredients are often mixed with excipients which are pharmaceutically acceptable and compatible with the active ingredient. An API can be formulated into the composition as neutralized pharmaceutically acceptable salt forms. Pharmaceutically acceptable salts include the acid addition salts (formed with the free amino groups of the polypeptide or antibody molecule), which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed from the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, 2- ethylamino ethanol, histidine, procaine, and the like.

[00150] Some examples of substances which can serve as pharmaceutically-acceptable carriers or components thereof are sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and methyl cellulose; powdered tragacanth; malt; gelatin; talc; solid lubricants, such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils, such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and oil of theobroma; polyols such as propylene glycol, glycerine, sorbitol, mannitol, and polyethylene glycol; alginic acid; emulsifiers, such as the Tween™ brand emulsifiers; wetting agents, such sodium lauryl sulfate; coloring agents; flavoring agents; tableting agents, stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline; and phosphate buffer solutions. The choice of a pharmaceutically-acceptable carrier to be used in conjunction with the compound is basically determined by the way the compound is to be administered. If the subject compound is to be injected, the preferred pharmaceutically-acceptable carrier is sterile, physiological saline, with a blood-compatible suspending agent, the pH of which has been adjusted to about 7.4.

[00151] In addition, the compositions may further comprise binders (e.g. acacia, cornstarch, gelatin, carbomer, ethyl cellulose, guar gum, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, povidone), disintegrating agents (e.g. cornstarch, potato starch, alginic acid, silicon dioxide, croscarmelose sodium, crospovidone, guar gum, sodium starch glycolate), buffers (e.g., Tris-HCL, acetate, phosphate) of various pH and ionic strength, additives such as albumin or gelatin to prevent absorption to surfaces, detergents (e.g., Tween 20, Tween 80, Pluronic F68, bile acid salts), protease inhibitors, surfactants (e.g. sodium lauryl sulfate), permeation enhancers, solubilizing agents (e.g., glycerol, polyethylene glycerol), anti-oxidants (e.g., ascorbic acid, sodium metabisulfite, butylated hydroxyanisole), stabilizers (e.g. hydroxypropyl cellulose, hyroxypropylmethyl cellulose), viscosity increasing agents(e.g. carbomer, colloidal silicon dioxide, ethyl cellulose, guar gum), sweeteners (e.g. aspartame, citric acid), preservatives (e.g., Thimerosal, benzyl alcohol, parabens), lubricants (e.g. stearic acid, magnesium stearate, polyethylene glycol, sodium lauryl sulfate), flow-aids (e.g. colloidal silicon dioxide), plasticizers (e.g. diethyl phthalate, triethyl citrate), emulsifiers (e.g. carbomer, hydroxypropyl cellulose, sodium lauryl sulfate), polymer coatings (e.g., poloxamers or poloxamines), coating and film forming agents (e.g. ethyl cellulose, acrylates, polymethacrylates) and/or adjuvants.

[00152] Dry powder compositions may comprise propellants such as chlorofluorocarbons 12/11 and 12/114, or, in another embodiment, other fluorocarbons, nontoxic volatiles; solvents such as water, glycerol and ethanol, these include co-solvents as needed to solvate or suspend the active; stabilizers such as ascorbic acid, sodium metabisulfite; preservatives such as cetylpyridinium chloride, benzalkonium chloride and phenoxy ethanol; tonicity adjustors such as sodium chloride; buffers; and flavoring agents such as sodium saccharin.

[00153] The compositions may also include incorporation of the active material into or onto particulate preparations of polymeric compounds such as polylactic acid, polglycolic acid, hydrogels, etc, or onto liposomes, microemulsions, micelles, unilamellar or multilamellar vesicles, erythrocyte ghosts, or spheroplasts.) Such compositions will influence the physical state, solubility, stability, rate of in vivo release, and rate of in vivo clearance.

[00154] Also comprehended by the invention are particulate compositions that may be coated with polymers (e.g. poloxamers or poloxamines).

[00155] In some embodiments, compounds modified by the covalent attachment of water- soluble polymers such as polyethylene glycol, copolymers of polyethylene glycol and polypropylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone or polyproline. The modified compounds are known to exhibit substantially longer half-lives in blood following intravenous injection than do the corresponding unmodified compounds (Abuchowski et al., 1981; Newmark et al., 1982; and Katre et al., 1987). Such modifications may also increase the compound's solubility in aqueous solution, eliminate aggregation, enhance the physical and chemical stability of the compound, and greatly reduce the immunogenicity and reactivity of the compound. As a result, the desired in vivo biological activity may be achieved by the administration of such polymer-compound abducts less frequently or in lower doses than with the unmodified compound. [00156] The following examples are presented in order to more fully illustrate certain embodiments of the invention. They should in no way be construed, however, as limiting the broad scope of the invention.

EXPERIMENTAL DETAILS SECTION

Example 1;

In- vitro trials on Corpus Cavernosum tissue and reduction of the response

[00157] A strip of rabbit corpus cavernosum was placed under 2 g tension in a 10 mL bath of Krebs solution pH 7.4 at 32°C and sub-maximal tonic isometrically recorded contraction is induced by phenylephrine HCl (3 μΜ). Test substance-induced isometrically recorded relaxation within 5 minutes by 50 percent or more (>50%), relative to the 0.3 μΜ sodium nitroprusside relaxation indicates significant relaxation activity (see tables 1 to 7). The relaxation percentage provides a direct measure for erection efficiency.

[00158] Then, after the down-curves of each element (total 6) were established, various combinations were tested.

[00159] Table 1: combination of L-Arginine and Moxisylyte HCl

[00160] It seems that the combination of L-Arginine and Moxisylyte HCl, did not improve the performance of either compound. [00161] Table 2: combination of L-Arginine and Isosorbide dinitrate

[00162] It seems that there is a partial additive effect in the combination of L-Ar; and Isosorbide dinitrate.

[00163] Table 3: combination of Hydralazine HCl and Moxisylyte HCl

[00164] It seems that the combination of Hydralazine HCl and Moxisylyte HCl, did not improve the performance of either compound.

[00165] Table 4: combination of Hydralazine HCl and Isosorbide dinitrate (ISDN) Hydralazine HCl Isosorbide Relaxation (%) (μΜ) dinitrate (μΜ)

0.03 — 13.0

0.03 0.03 18.5

0.1 0.03 45.5

— 0.03 17.5

0.1 — 21.5

Note: Synergistic effect.

Table 4A: combination of Hydralazine HCl, Moxisylyte and Isosorbide dinitrate

[00167] Surprisingly, it seems that there is a synergistic effect in the combination of Hydralazine HCl and Isosorbide dinitrate and Moxisylyte and ISDN.

[00168] Table 5: combination of L-Arginine and Hydralazine HCl

[00169] It seems that the combination of L-Arginine and Hydralazine HCl, did not improve the performance of either compound.

[00170] Table 6: combination of Isosorbide dinitrate and Moxisylyte HCl

[00171] Surprisingly, there is a full additive effect in the combination of Isosorbide dinitrateand and Moxisylyte HCl.

[00172] Table 7: combinations of: (1) Hydralazine HCl, Moxisylyte HCl and L-Arginine; and (2) Hydralazine HCl, Moxisylyte HCl, L-Arginine and L-Norvaline

[00173] Table 7 demonstrates that the addition of either L-Arginine to the combination of Hydralazine HCl and Moxisylyte HCl or L-Arginine and L-Norvaline to the combination of Hydralazine HCl and Moxisylyte HCl had an additive effect in this experiment.

[00174] Table 7: combinations of Hydralazine HCl, Isosorbide dinitrate and L-Arginine; and (2Hydralazine HCl and Isosorbide dinitrate, L-Arginine and L-Norvaline

[00175] Table 8 demonstrates that the addition of either L-Arginine to the combination of Hydralazine HCl and Isosorbide dinitrate or L-Arginine and L-Norvaline to the combination of Hydralazine HCl and Isosorbide dinitrate had at least an additive effect in this experiment.

[00176] Table 9: combinations of Moxisylyte HCl, Isosorbide dinitrate and L-Arginine; and (2 Moxisylyte HCl and Isosorbide dinitrate, L-Arginine and L-Norvaline

[00177] Similarly to tables 7 and 8, table 9 also demonstrates that the addition of either L- Arginine to the combination of Moxisylyte HCl and Isosorbide dinitrate or L-Arginine and L-Norvaline to the combination of Moxisylyte HCl and Isosorbide dinitrate had at least an additive effect in this experiment.

Example 2:

Combinations across healthy human penile skin and mouse skin

[00178] The aim of this study was to perform in vitro skin drug permeation testing from 6 formulations with and without the penetration enhancers using human penile epidermal membrane.

[00179] Figures 1 and 2 show the cumulative amount of drug per cm ² permeated across human penile epidermal membrane after t=6 hours and 24 hours, respectively. Higher levels of Isosorbide dinitrate (ISDN) were detected in the receiver fluid with no obvious difference with the inclusion of a penetration enhancer. Moxisylyte without penetration enhancer was detected at concentrations similar to ISDN, however the saturated solution containing a penetration enhancer was observed to have negligible levels of drug present in the receiver fluid after t=6h. After t=24h there was a marked increase in the levels of ISDN, Moxisylyte and Hydralazine compared to the same solutions at the t=6 h time point. After 24 the highest delivery was observed from ISDN solutions with and without a penetration enhancer, followed by samples containing Moxisylyte and Hydralazine.

[00180] In a clinical setting the delivery of drugs to the target condition is not the only issue and relative potencies need to be considered. As such a predicted efficacy (putative efficacy index, PEI) for each compound was calculated using the flux at 24 h and the IC50 values (potencies) (Table 10). The resultant data is presented for the single drug solutions in Figure 3. Based on the calculated PEI the top ranked compound is Moxisylyte primarily as a result of its high potency. Obviously caution needs to be taken when reviewing this data as a result of the variability observed. Nevertheless figure 3 does suggest that the order of preference for the drugs would be would be: Moxisylyte > ISDN>Hydralazine

[00181] Table 10: Mechanism of action and potency of candidate drugs

[00182] The flux and PEI for all solutions tested was calculated at the t=24 hours time point and the results are summarized in Table 11 and 12, respectively. The flux data (Table 11) provides evidence of a potential synergistic effect between the ISDN and Hydralazine where in the presence of Hydralazine, ISDN was observed to have higher flux (compared to ISDN alone); however, the reverse trend was not observed for Hydralazine in the presence of ISDN.

[00183] For the reason discussed above the putative efficacy index (PEI) for each compound was calculated using the flux at 24hours and potencies supplied by the Sponsor. The additive potency of the combination solutions was then calculated and the results are summarised in Table 12. The additive potency value is not representative of in vivo efficacy, but instead has been used to rank the potential potency of combination formulations. The solutions ranked 1 to 6 all contain Moxisylyte (either individually or in combination). ISDN and Hydralazine in combination were ranked 7th and 8th (without and with penetration enhancer, respectively) followed by solutions containing ISDN alone ranked 9th and 10th (without and with penetration enhancer, respectively) and finally solutions containing Hydralazine alone ranked 11th and 12th (with and without penetration enhancer, respectively).

[00184] Table 11: Flux ^g/cm2 h-1) calculated from t=24 h. Mean + SEM (n=4-6).

ISDN / Hydralazine DF 1 17.93 + 9.94 - 3.19 + 2.70

ISDN / Hydralazine DF 2 15.36 ± 9.17 - 2.18 + 1.44

ISDN / Moxisylyte DF 1 3.94 + 3.08 0.77 + 0.74 -

ISDN / Moxisylyte DF 2 9.02 + 6.78 0.75 + 0.53 -

Hydralazine / Moxisylyte DF 1 - 0.70 + 0.60 2.50 + 2.23

Hydralazine / Moxisylyte DF 2 - 0.32 + 0.25 9.54 + 7.76

Table 12: Calculated putative efficacy index for drug and drug combinations

[00186] The selection of a single compound formulation appears to be a clear choice, where due to the low IC50 Moxisylyte would be the most promising candidate. If a combination product is preferential then then a variety of factors should be considered. A combination product is most relevant when the two or more drugs have different mechanisms of action, ideally producing a synergistic effect. Fortunately this is the case for ISDN and Moxisylyte (ISDN being an NO donor and Moxisylyte acting as an al- adrenergic receptor antagonist), suggesting that a combination product utilizing these two drugs may be an interesting prospect. There is less of a case for combining ISDN and Hydralazine as both are NO donors and it is unclear how much of the action of Hydralazine is related to its K+ channel activator action. Furthermore during the feasibility experiments Hydralazine was observed to have poor stability in a selection of receiver fluids, possibly indicating that this compound may provide a more challenging formulation development pathway. Based on this information the recommended order of preference for a combination product is shown below:

ISDN with Moxisylyte > Moxisylyte with Hydralazine > Hydralazine with ISDN.

[00187] The data generated during the in vitro permeation supports this line of reasoning; furthermore the concentrations of drug detected in the receiver fluid at 24 hours were markedly above the IC50 values provided by the Sponsor, further supporting the potential of a successful formulation.

[00188] Table 13: Calculated concentration of each compound at t=24 h.

Hydralazine DF 2 - - 76.79

ISDN / Hydralazine DF 1 668.31 - 142.67

ISDN / Hydralazine DF 2 572.41 - 97.63

ISDN / Moxisylyte DF 1 146.70 21.41 -

ISDN / Moxisylyte DF 2 336.18 20.99 -

Hydralazine / Moxisylyte DF 1 - 19.59 111.89

Hydralazine / Moxisylyte DF 2 - 8.94 427.15

[00189] Further studies performed with nude mice as it is the optimal specie for vasodilator studies. Blood flow on animal back was measured by a Laser Doppler Pulsar at 10 and 40 min. after compound treatment (ISDN, Hydralazine, and Moxisylyte at various dosages, concentrations, combinations, and formulations as described hereinabove).

[00190] Table 14: the formulation utilized in the mice studies

Mineral oil 8 8 8 8 8 8 8 8

White Pet 2 2 2 2 2 2 2 2

Cetyl 4 4 4 4 4 4 4 4 Alcohol

GMS 1 1 1 1 1 1 1 1

DMI 10 10 10 10 10 10 10

ISDN 1 1 1

Hydralazine 0.5 0.5 0.5

Moxisylyte 0.1 0.1 0.1

[00191] Table 15: Selection of preferred combinations using Laser Doppler Pulsar

Weegum 1.0

Xanthan gum 1.0

total 100 100 100 100 100.0 100

[00192] Table 15: depicts improved formulations with higher APIs concentration, better penetration system and more sensitive detection system (laser Doppler).

[00193] The animal back of the mice was divided. The right side from the spinal cord received 50 μΐ of the tested compound. The Left side was clear and was used as control.

[00194] Twenty four hours after the treatments the animals were observed for signs of irritations. All the treated areas of the animals were normal and no signs of irritation were observed. The flow and diameter parameters of the vehicle group were nearby 100% after 10 and 40 minutes, indicating no changes during the examination. At ten minutes post treatment, the Blood Flow % of the tested compounds increased between 300-400% and the Vessel diameter percent increases also between 150-200%. After 40 minutes, the blood flow % (Fig. 6) and vessel diameter % of the tested compounds (Fig. 7) and the positive control (Rectogesic) increased to 600% and 200% respectively. All the active compounds and positive control treatments were statistically significant different (p<0.001) from the Vehicle group.

[00195] According to these experiments assessing the vasodilator effect of different combinations (tables 14 and 15) of active compounds with Hydralazine, Moxisylyte, Phentolamine, and NaOH 5%aq, the major differences between the active compounds and the positive control, Rectogesic were observed after the first ten minutes post dosing (Figs. 6 and 7). The skin absorption of the positive control Rectogesic, was faster and induced a significant increase in blood flow and vessel diameter compared to the control vehicle group. [00196] Faster increase in blood flow was recorded in the presence of Phentolamine. After 40 minutes, all the tested compounds achieved the same change in blood flow and vessel diameter.

Example 3:

Formulation development of separate single formulations

[00197] During the initial pre-formulation experiments poor chemical stability was observed when both ISDN and Moxisylyte were combined. As a result Moxisylyte and ISDN were formulated as 2 individual products rather than a dual combination product (i.e, containing both Moxisylyte and ISDN together). The solubility of Moxisylyte and ISDN was individually assessed in solvent systems that would form the basis of a solution or gel.

[00198] Due to further compatibility and stability drawbacks a kit comprising a powder formulation and a separate topical formulation was further developed.

[00199] Initial experiments were performed in order to assess the feasibility of incorporating Moxisylyte or Hydralazine as a powder into various ISDN formulations. The ISDN formulations selected were prepared without the inclusion of the gelling agent (HPC, HPMC or HEC) and the solubility of Moxisylyte and Hydralazine assessed. Into each solvent system containing ISDN, 1% w/w of Moxisylyte or Hydralazine was added (as a powder), the vial shaken by hand and the time taken to dissolve recorded.

[00200] The results show that Moxisylyte was more soluble than Hydralazine in all systems at the concentration assessed. In addition it would appear that both Hydralazine and Moxisylyte were more soluble in the aqueous systems in comparison to the non-aqueous systems which is most likely due to the water present in the aqueous system as high solubility was observed for both Hydralazine and Moxisylyte in water (> 3.90 - 5.76 and > 8% w/w, respectively). It is thought that the time taken for Moxisylyte to dissolve in the aqueous systems would be acceptable for a dual compound formulation where Moxisylyte was stored as a powder and ISDN was present in a solvent system.

[00201] It is possible that the water content could be increased in the aqueous formulations to increase the solubility of Moxisylyte and Hydralazine in the formulation and therefore increase their dissolution rate, however as ISDN is poorly water soluble the concentration of ISDN present in the formulation would need to be decreased. Alternatively a lower concentration of Moxisylyte or Hydralazine could be incorporated into the formulation which again should speed up the dissolution rate.

[00202] Solvent systems were developed with increased levels of water and incorporating excipients where Hydralazine has previously shown good solubility. The solubility of ISDN and Hydralazine was assessed in these developed systems in addition to one of the current

ISDN formulations to be able to compare the solubility. The results show that the solubility of Hydralazine increased with the increased water content, however a decrease in the solubility of ISDN was observed. All of the developed solvent systems were observed to have higher Hydralazine solubility.

Example 4:

Stability of Moxisylvte/ISDN in Buffers

[00203] Following the API/excipient stability experiments the stability of Moxisylyte and

ISDN in buffers with and without β-cyclodextrin was investigated where solutions containing 0.1% w/w Moxiylyte and/or ISDN were placed on stability at 40°C and 50°C.

[00204] The results suggested that Moxisylyte alone appeared to be more stable at lower pH values, where the percentage recovery following 1 week storage at 50°C of Moxisylyte was observed to be 94.75 and 92.09% at pH 4 and 5, respectively compared to 78.56 at pH

6. This trend was also observed for ISDN alone where the percentage recovery of ISDN at pH 4, 5 and 6 stored at 50°C for 1 week was 99.18, 98.74 and 90.83%, respectively. In these buffer systems (pH 4, 5 and 6), where Moxisylyte and ISDN were stored in combination, the percentage recovery of Moxisylyte was similar to its storage alone, suggesting that ISDN did not have an effect on Moxisylyte stability in these systems. However a decrease in recovery of ISDN was observed in these systems when stored with Moxisylyte, which indicates that Moxisylyte may have an affect on the stability of ISDN.

[00205] When compared to the buffers alone, degradation was also observed to increase for both Moxisylyte and ISDN stored in combination and alone when β-cyclodextrin was incorporated into the buffer. In addition Moxisylyte and ISDN remain relatively stable after 1 week storage at 50°C when stored alone in 50:50 ethanohcyclomethicone (% recovery = 98.96 and 98.85%, respectively), however when these compounds stored in combination in this system a decrease in percentage recovery was observed for both Moxisylyte (89.68 %) and ISDN (86.09%).

[00206] To reduce the risk of any combination product having stability issues a Dual Packaging System, such that ISDN and Moxisylyte would only be combined upon application and the formulation development performed was investigated.

Example 4:

Dual Formulation Stability

[00207] Several formulations combining ISDN and Moxisylyte were prepared and examined. All formulations were assessed under light microscope at 200x and 400x magnification and compared to their respective placebo formulations. Crystallisation was not observed in any of the active formulations at t=0 or following 4 week storage at 25 °C and 40°C.

[00208] The formulations were visually assessed. At t=0, where all active and placebo non-aqueous ISDN and Moxisylyte gels were observed to be slightly opaque viscous gels and the ISDN and Moxisylyte solvent system formulations were observed to be colorless, low viscosity liquids. The aqueous active and placebo ISDN gel formulations were observed to be colorless viscous gels. Following 4 weeks storage at 25°C and 40°C all formulations were observed to be unchanged from t=0.

Incorporation of Powdered Moxisylyte into ISDN Formulation

[00209] Experiments were performed in order to assess the feasibility of incorporating Moxisylyte as a powder into ISDN formulations on stability. The ISDN formulations selected for stability were prepared without the inclusion of the gelling agent and the solubility of Moxisylyte was assessed. Into each solvent system containing ISDN, 1% w/w of Moxisylyte was added (as a powder), the vial shaken by hand and the time taken to dissolve recorded. The results are summarized in Table 11. In addition it would appear that Moxisylyte is more soluble in the aqueous systems in comparison to non-aqueous systems, which is most likely due to the water present in the aqueous system as high solubility was observed for Moxisylyte in water (> 8% w/w). It is thought that the time taken for Moxisylyte to dissolve in the aqueous systems would be acceptable for a dual compound formulation where Moxisylyte was stored as a powder and ISDN was present in a solvent system.

[00210] Table 16. Time taken for Moxisylyte as a powder in the developed ISDN solvent systems to dissolve (seconds)

FIN 1 > 900

FIN 2 > 900

FIN 4 Approx. 600

FIN 5 Approx. 420

FIN 6 Approx. 300

[00211] Selected formulations (table 17) were chosen based on the fast dissolving of Moxisylyte in this ISDN-containing formulation.

in vitro Drug Release

[00212] Full scale in vitro drug release experiments were performed on 10 formulations. The formulations were selected based on the pre-formulation, formulation development and in-use stability experiments performed.

[00213] The in vitro drug release experiments were performed using Franz Cell as illustrated in Fig. 22. These in vitro skin permeation experiments involved the use of a diffusion cell designed to mimic the physiological and anatomical conditions of skin in situ. Synthetic membrane or human epidermal penile skin (including the subcutaneous tissue and connective tissue) were positioned between the two halves with the Stratum corneum (upper) facing the donor compartment allowing for drug application.

[00214] The aim of the work was to compare each drug concentration permeating across human penile skin when applied from different test systems to identify a lead drug/drug combination candidate.

[00215] The full scale in vitro drug release investigation was performed across PTFE membrane, where a total of 10 formulations containing ISDN were assessed. Sampling time points were taken at 30 min, 1, 2, 3, 4, 6, 24 and 30 h and the results of the drug release investigation are presented in Fig. 8, in which the mean cumulative amount of ISDN released per unit area against time is illustrated.

[00216] A steady state was achieved for all formulations between 30 min and 4 h. The formulations were ranked according to their respective drug steady state. The release of ISDN appeared to be concentration dependent. In general, the formulations containing a lower concentration of Transcutol P were observed to have lower release rates of ISDN than the formulations containing higher Transcutol P concentration.

[00217] The full scale in vitro drug release investigation was performed on a total of 3 formulations containing Moxisylyte across PTFE membrane as illustrated in Fig. 9. A steady state was calculated between 30 min and 4 h. The formulations were ranked according to their respective drug steady state. Thus, a relatively high amount of ISDN was observed to be released from all of the formulations tested. Past ISDN formulations closer to saturation (approximately 90% of saturated solubility) are inferior to formulations wherein Moxisylyte is incorporated at 1 w/w to ensure the dissolution time is less than 1 min. In addition, higher drug concentration of ISDN (between 1 and 2% w/w) is present in the majority of formulations compared to Moxisylyte (1 % w/w).

Selected formulations

[00218] Selected formulations and a placebo are detailed in Table 17.

[00219] Table 17. Selected formulations

Transcutol P 5-100 5-100

PEG 400 5-100 5-100

Propyl paraben 0.05-0.8 0.05-0.8

Methyl paraben 0.005-0.01 0.005-0.01

Moxisylyte 0.2-5

Total 100.000 90-100

[00220] The ready to use formulations were a transparent liquid that can be readily applied locally.

[00221] The ready to use formulations were stored at room temperature.

[00222] ISDN and Moxisylyte analysis was performed using HPLC as detailed in Table 18.

[00223] Table 18. ISDN and Moxisylyte HPLC analysis.

Needle Wash 80:20 Deionised water / HPLC grade acetonitrile

Pump Wash 60:40 Deionised water / HPLC grade acetonitrile

The following parameters were assessed over the stability studies performed: ISDN/Moxisylyte content;

ISDN/Moxisylyte peak purity and related substances;

Visual appearance (macroscopic / microscopic); and

• Time to dissolve Moxisylyte in the ISDN solution. Dual Packaging System

[00225] Due to the poor stability of Moxisylyte in solvent systems it was decided to use a Dual Packaging System in a way that ISDN and Moxisylyte would only be combined upon application.

[00226] Two systems were used as detailed below by the medical personnel or by the patient as a friendly and easy-to-use unit.

Preparation by Medical Personnel

[00227] This system was involved transfer of the ISDN solution from one bottle to the bottle containing the Moxisylyte powder.

Dual Pouch Tube

[00228] The Dual Pouch Tube: does not allow the solution (3 mL of ISDN solution) to be in contact with the powder (30 mg Moxisylyte) before the user breaks the divider between the two pouches. Upon breaking the divider, the solution and powder were mixed and the powder was being dissolved in the solution within seconds. The user was instructed to use the content within few hours following mixing and not more than 12 hours. [00229] The selected formulations (table 17) were transparent, lightly viscous fluid that was applied with a single glove on the penis. The remains were washed with water or rinsed with pre-moistened towel before intercourse.

[00230] The selected formulations (table 17) did not cause any adverse events, including skin irritation, skin sensitization and no change in arterial blood pressure.

[00231 ] The major differences between the active compounds were observed after the first two hours post dosing. The skin absorption of the all test items, was fast and induced a significant increase in blood flow and vessel diameter compared to the control vehicle group within the first 10 minutes post dosing.

[00232] Blood flow and blood vessel diameter parameters of the placebo groups were close to 100% following all observation periods up to 240 minutes, indicating no change during examination. At five minutes post treatment, the Blood Flow % of the tested selected formulations (table 17) unexpectedly increased between 150-200% and the Vessel diameter percent also increased between in the range 110-132%.

[00233] Between 30 and 120 minutes, the percent blood flow and percent vessel diameter of the tested compounds increased to the range -500% and -250%, respectively. All the active compounds treatments exhibited statistically significant difference (p<0.001) compared to the vehicle group. Individual blood flow measurements were found to correlate nicely with the measurements of vessel diameter.

[00234] In the ex vivo set of trials male albino New Zealand (NZ) rabbits of 4-8 months old were used. The animals were sacrificed and the corpus cavernosum was isolated. A strip from the basal area of the corpus cavernosum was removed and placed under 2-g tension in a 10-mL tissue bath containing Krebs solution pH 7.4 at 32°C. Sub-maximal isometrically recorded tonic contractions are induced by phenylephrine (3 μΜ). Test substance (30 μΜ)- induced relaxation by 50% or more (>50%) within 5 min, relative to the effect of 0.3 μΜ sodium nitroprusside, indicates significant relaxation (Angulo, J., Cuevas, P., Moncada, I., Martin-Morales, A., Allona, A., Fernandez, A., Gabancho, S., Ney, P. and Saenz de Tejada, I. Rationale for the combination of PGE(l) and S-nitroso-glutathione to induce relaxation of human penile smooth muscle. J. Pharmacol. Exp. Ther. 295: 586-593, 2000.).

[00235] Results are detailed in Table 19.

[00236] Table 19. The effect of ISDN and Moxisylyte on the relaxation of isolated rabbit Corpus cavernosum pre-contracted with phenylephrine.

0.3nM the relaxation reaches 37.0% and 44.5%, respectively. It should be noted that these data present an additive effect since the relaxation caused by 0.1 nM or 0.3nM Moxisylyte alone is 16.5% and 28.0%, respectively.

[00238] The additive effect of ISDN and Moxisylyte is not surprising based on the different mechanism of action of each.

[00239] Additionally, no toxic signs, pre-terminal deaths or skin reactions observed and no abnormalities detected at necropsy. Based on the results of the present study the test item cannot be classified as skin irritant (Regulation (EC) No 1272/2008 of the European Parliament and of the council of 16 December 2008 on classification, labelling and packaging of substances and mixtures, amending and repealing Directives 67/548/EEC and 1999/45/EC, and amending Regulation (EC) No 1907/2006; OECD: Harmonized Integrated Classification system (OECD, 2001).) or Sensitizer (as per OECD: Harmonized Integrated Classification system (OECD, 2001).

[00240] No blood penetration: in these PK trials, the rabbit's ears were treated topically by the test items. The dose volume administrated was 200 μΐ. Blood samples (approximately 1 ml of whole blood) were collected at the following time points: 0 (baseline), 5, 10, 30, 60, 90, 120 and 240 min after compounds application. The blood samples, containing EDTA, were centrifuged (5000 rpm, lOmin, 4°C) for plasma separation and the plasma was transferred to clean vials, labeled with the study code, rabbit number and time-point. All plasma samples were frozen immediately at -20°C and treated with Paraoxon-ethyl [CAS Number 311-45-5] as an esterase inhibitor. Moxisylyte was analyzed from LLOQ (Lower limit of quantification) of 1 ng/mL and ISDN from 5 mg/mL. No detectable concentrations of ISDN neither Moxisylyte were analyzed is all samples tested, including placebo.

[00241] Effect of the selected formulations (table 17) on Arterial Blood Pressure: Arterial blood pressure was examined in 16 rabbits, as detailed above (Trial C). Mean arterial blood pressure was measured by Biopac system before test items application and at 30, 60, 120 and 240 min after compounds treatment. No change was found in the mean arterial blood pressure during all measurements.

[00242] Efficacy of VL#FIA3-30. First trial: The flow and diameter parameters of the vehicle mice group (n=12) were nearby 100% after 10 and 40 minutes, indicating no changes during the examination. At ten minutes post treatment, the Blood Flow % of the selected formulations (table 17) were (entitled: 38) increased between 300% - 400% and the Vessel diameter percent increases also between 150% - 200%. After 40 minutes, the blood flow % and vessel diameter % of the tested compounds and the positive control (Rectogesic) increased to 600% and 200%, respectively. All the active compounds and positive control treatments were statistically significant different (p<0.001) from the vehicle group as illustrated in Fig. 10 (blood flow) and Fig. 11 (blood vessel diameter).

[00243] Second trial: Blood flow and blood vessel diameter parameters of the vehicle rabbit group (n=16) were nearby 100% after up to 240 minutes, indicating no changes during the examination. At thirty minutes post treatment, the blood flow % of the tested compound increased by -500% and the blood vessel diameter percent increases also -250% (Fig. 12 and Fig. 13, respectively). All the active compounds and positive control treatments were statistically significant different (p<0.001) from the Vehicle group. Individual blood flow measurements were found to correlate nicely with the measurements of vessel diameter, as can be seen in the example for the 60 minutes measurements of the treated groups in Fig. 14. The tested selected formulations (table 17) seemed to induce a faster and more prominent increase in blood flow and vessels diameter.

[00244] VL#FIA3-30: Human skin irritation potential Clinical Study: Treatment sites were assessed according to the scoring scale of Annex V Directive 67/S 18/EEC for erythema and oedema. The incidence and extent of exudation or surface encrustation and discoloration of the treated sites were also assessed. Any other effects and subjective comments made by the volunteers were recorded. A Preliminary Study was performed on three volunteers to reduce the likelihood of excessive skin irritation. The Preliminary Study was performed using exactly the same ethical and methodology conditions as the Main Study. 10 volunteers were participating in the Main Study with a range of ages of 25 - 72 and an average age of

49 years. No subjects discontinued or missed any application or reading. In both trials, 1cm x 1 cm samples of the Test Material were applied to the back of a patch test, Haye's Test

Chambers. Each volunteer was patched in two sites on the back with the same Test Material. One hour after patching, one patch was removed and the skin was graded immediately, one hour and 24 hours after patch removal. Since no grades were observed for the one-hour patch, the other patch was left in place for a total of 4 hours and the skin was graded after removal. The average erythema score (24/48/72 hours) was: 0 (no erythema). The average oedema score (24/48/72 hours) was: 0 (no oedema). No skin reactions were observed in all volunteers during the studies for erythema, oedema and exudation; i.e., one hour after removal of the 1 hour application patches, one hour after removal of the 4 hours application patches, 24 hours after patch removal, 48 hours after patch removal and 72 hours after patch removal. Example 5:

Activating a refractory clinical dose of a PDE5-inhibitor

[00245] This experiment involved subjects who suffered from ED for more than 6 months with stable heterosexual relationship with the same partner, their International Index of Erectile Function- Erectile Function (IIEF-EF) baseline score was from 11 to 19. These subjects faced at least 50% unsuccessful sexual intercourse for 4 attempts on 4 different days and they were aged between 45 and 67.

[00246] Prior to the trial they used PDE5 -inhibitor (Viagra) with very limited and unsatisfying response. During the trial, they didn't use any medication dedicated to treated ED, including PDE5 -inhibitors, for the duration of the washout period (10-14 days), the trial (up to 7 days) and the follow-up period (7 days), total of up to 28 days.

[00247] During the trial, they reported some efficacy of the VL#FIA3-30 preparation. After the completion of the trial, they re-used PDE5 -inhibitors and reported extremely strong and satisfying erections, in a way they have never experienced before. [00248] It will be appreciated by a person skilled in the art that the present invention is not limited by what has been particularly shown and described hereinabove. Rather, the scope of the invention is defined by the claims which follow: