Patent Application
20090118352
The present invention relate to the topical application of a novel composition containing N-cocoyl amino acid pyrrolidone salts for preventing or reducing the transmission of sexually transmitted diseases (STDs) in sexually active individuals. This invention also relates to compositions and methods for preventing conception and/or reducing the risk of conception in sexually active females. The present compositions and methods are especially effective for preventing of sexually transmitted diseases during male/female sexual intercourse. It can, however, be used by heterosexual, homosexual, and bisexual individuals to reduce the risk of being infected by, or of transmitting, a sexually transmitted disease through sexual contact.
The method of this invention comprises application of an effective amount of the microbicidal formulation within the vagina or anorectic region with choice of different dosage form applications. The invention also relates to methods of inhibiting the activity of other pathogens whose mode of transmission is nonsexual. The compositions of use in the invention are also effective in the inhibition of bacteria and fungi, which coexist with viruses or viral infections.
Sexually transmitted diseases (STDs) are among the most prevalent and communicable diseases, and continue to be a significant public health problem. It is estimated that more than 250 million people worldwide, and close to 3 million people in the United States, are infected annually by gonorrhea. Annual worldwide incidence of syphilis is estimated at 50 million people, with 400,000 in the United States annually needing treatment. The human immunodeficiency virus (HIV), resulting in fatal acquired immunodeficiency syndrome (AIDS), has spread rapidly in both homosexual and heterosexual groups. For many sexually transmitted infections, vaccines do not exist, and therapeutic agents are only partially effective, expensive, and difficult to distribute. In addition, female partners in many relationships do not control pregnancy or STI. One approach to the general control of STDs is the use of topically applied, female controlled microbicides that inactivate the relevant pathogens.
It is thought that certain bacteria known to cause STD's may aid in HIV transmission. In persons who have been exposed to HIV, certain bacteria that cause STD's often fail to respond to therapies that are otherwise highly effective. HIV infection may help the spread of a bacterial STD that in turn helps to spread HIV. STD's pathogens such as Chlamydeous, syphilis, genital herpes and gonorrhea that cause ulcerations of the genital skin seem to increase the risk of acquiring or transmitting HIV infection sexually.
NONOXYL-9 (N-9) is present in a large number of condoms and other spermicidal agents. Recent research, however, has found that N-9 can disrupt epithelial cells in the vagina, thereby increasing the risk of HIV infection. Frequent use of N-9 is also positively correlated with bacterial vaginosis, genital ulcers and Vulvitis, vaginal Candidiasis, toxic shock syndrome, and epithelial disruption of the cervix and the vagina. The final phase III multi-centre randomised placebo-controlled trial of N-9, undertaken by the United Nations Joint Programme on HIV/AIDS in 2002, showed that N-9 had no efficacy in preventing HIV transmission. Indeed, the transmission rate was marginally higher in the N-9 group. In 2003 the World Health Organization (WHO) recommended that people not use N-9 for protection against HIV/AIDS or other STIs. (Van Damme L, et al. (2002) Effectiveness of COL-1492, a nonoxynol-9 vaginal gel, on HIV-1 transmission in female sex workers: A randomised controlled trial. Lancet 360:971-977.; Stephenson J (2000) Widely used spermicide may increase, not decrease, risk of HIV transmission. Jama 284(8): 949.)
U.S. Pat. No. 5,004,757 is directed to a method of deactivating viruses on surfaces by applying a three-part composition containing glutaraldehyde. The composition also contains hydrogen-bonded glycol molecules to eliminate aldehyde odor, and an anionic surfactant such as sodium dodecyl sulfate (SDS) as a potentiator of the virucidal activity of the glutaraldehyde component. The patent indicates that SDS has limited virucidal activity on its own, but has a synergistic effect when combined with gluteraldehyde. Due to the fact that gluteraldehyde is a well-known mutagen, the invention is not useful with direct contact of human epithelium tissues.
U.S. Pat. Nos. 6,063,773, 7,078,392 presented a method for treating and preventing various infections, including papilloma virus and fungal and parasitic infections. The inventions relate to use sulfated polysaccharide, such as cellulose sulfate (a high molecular weight carboxymethyl cellulose-based polymer), for treating and preventing various sexual infections. The cellulose sulfate formulations were tried clinically for preventing HIV transmissions. In 2007, World Health Organization and UNAIDS has prematurely stopped the clinical trial of using cellulose sulfate to prevent HIV transmission in women because of a higher number of HIV infections in the active group compared with the placebo group. (Lut Van Damme et al. Lack of Effectiveness of Cellulose Sulfate Gel for the Prevention of Vaginal HIV Transmission, N Engl J med 359:463-472; Statement by the world Health Organization and UNAIDS: Cellulose sulfate microbicide trial stopped, 31 Jan. 2007)
U.S. Pat. Nos. 5,314,917 and 6,297,278 provided an antimicrobial composition for inhibiting the activity of enveloped viruses, treatment of viral infections and method for inactiviting sperm. The composition comprises: a mixtures of a betaine, an amino oxide and a protonating agent to adjust the pH of the overall composition to about 5.5 or below (C31G). Clinical trials showed that C31G was associated with higher reporting of reproductive adverse events. The data were insufficient to conclude whether C31G is effective at preventing HIV infection relative to placebo. (Feldblum P J, et al. (2008) SAVVY Vaginal Gel (C31G) for Prevention of HIV Infection: A Randomized Controlled Trial in Nigeria. PLoS ONE 3(1): e1474. Peterson L, et al. (2007) SAVVY(R) (C31G) Gel for Prevention of HIV infection in Women: A Phase 3, Double-Blind, Randomized, Placebo-Controlled Trial in Ghana. PLoS ONE 2(12): e1312.)
U.S. Pat. No. 5,617,877 introduced an acidic buffer gel contained within a flexible dome device positioned over the uterine cervix for controlling pH in the vagina for in-activating both sperm and STIs. The device is made of a dome and the absorbent acidic buffer gel is contained within the dome. The buffer gel (BufferGel™) is reported to be a negatively charged, non-absorbable high molecular weight polymer gel, designed to maintain vaginal pH below 5 in the presence of semen. BufferGel relies on a polymer comprised of carboxylated monomers to control the vaginal pH. BufferGel is placed over the uterine cervix that allowing the STD-causing microbes to readily migrate throughout the lower genital tract. The use of devices in combination with BufferGel requires significant skill and motivation by the user to obtain, and maintain, proper placement of the device. It is important to mention that some of the pathogens will survive the low pH environment. The efficacy of the acidic gel is solely depends on the acidity of the gel, and it might not guarantee a desirable effect on all the STI pathogens.
U.S. Pat. Nos. 7,226,914 and 7,235,536 relates to the treating, inhibiting or preventing of certain infectious agents including papilloma virus and various vaginitis-causing microbes by employing cellulose sulfate and other sulfated polysaccharides. Carraguard is used as a microbicidal gel containing a sulfated polysaccharide. Resent clinical trial with this sulfated polysaccharides composition as microbicide did not demonstrate that the compound were effective in preventing male-to-female HIV transmission during vaginal intercourse (Microbicide Trials Network: Statement on Carraguard phase III clinical trial findings, Feb. 18, 2008).
U.S. Pat. No. 6,417,144 relate to a solution for cleaning contact lenses. The solution comprises an amino acid derivative and at least one nonionic surfactant. The amino acid derivative is used as a component to increase the cleaning power together with a nonionic surfactant. The solution for contact lenses has an excellent cleaning effect and an adequate safety for the eye. This invention did not mention of preventing HIV and STDs.
The present invention relates to a novel composition and method for the prevention and control of sexually transmitted diseases and for the prevention of pregnancy. The invention also relates to treatment of viral infections. More particularly, the invention relates to a method for inhibiting the development of diseases and infections caused by viruses and some sexually transmitted pathogens whose major mode of transmission is sexual. In other aspects the invention also relates to methods of inhibiting the activity of enveloped viruses and other pathogens whose mode of transmission is nonsexual. The compositions of use in the invention are also effective in the inhibition of bacteria and fungi, which coexist with viruses or viral infections. Additionally, the invention relates to a prevention or treatment for virus related diseases, particularly sexually transmitted diseases related to AIDS, and to diseases related to this and other opportunistic infections of the immune-compromised host.
Transmission of HIV is often associated with the co-transmission of other viral and/or microbial pathogens. HIV may not be the sole agent responsible for AIDS (Duesberg, P. H. (1991) Proc. Natl. Acad. Sci. 88:1575-1579; Lemaitre, M., Guetard, D., Henin, Y., Montagnier, L. and Zerial, A. (1990). Res. Virol. 141:5-16). For this reason, antimicrobial agents, such as those described in this invention, with a broad spectrum of activities against viruses, bacteria, and yeasts may be of particular value in the prevention and treatment of Acquired Immune Deficiency Syndrome (AIDS). It is thought that certain bacteria known to cause STD's may aid in HIV transmission. In persons who have been exposed to HIV, certain bacteria that cause STD's often fail to respond to therapies that are otherwise highly effective. HIV infection may help the spread of a bacterial STD that in turn helps to spread HIV. STD's pathogens that cause ulcerations of the genital skin seem to increase the risk of acquiring or transmitting HIV infection sexually. The compositions described in the invention may also be of use to inactivate other viruses, including vaccinia, varicella, herpes zoster, cytomegalovirus, influenza, mumps and measles.
The present invention provides a novel composition and method for the prevention and control of sexually transmitted diseases and for the prevention of pregnancy. The invention also relates to treatment of viral infections. More particularly, the invention relates to a method for inhibiting the development of diseases and infections caused by viruses and some sexually transmitted pathogens whose major mode of transmission is sexual. In other aspects the invention also relates to methods of inhibiting the activity of pathogens whose mode of transmission is nonsexual. The compositions of the invention are also effective in the inhibition of bacteria and fungi, which coexist with viruses or viral infections.
The compositions can be formulated in the form of a gel, liquid, aerosol, mist, sponge, spray, foam, gel, cream, salve, jelly, suppository and film. The compositions can be applied to the external genital organs, vagina, anorectic region and rectum in different dosage forms with appropriate apparatus, for example the gel could be applied to vagina or rectum through an applicator or syringe. The compositions of the invention can also be incorporated into douches. The compositions can also be incorporated into wipes.
The invention also provides a method for preventing and the treatment of certain skin diseases. Such as ringworm and other fungi infectious skin diseases.
The invention also provides a broad spectrum and highly efficient microbicidal composition used for reducing the risk of transmission of STD-causing organisms to health care providers and laboratory personnel (or other persons) who may come in contact with biological samples and specimens
The compositions of the present invention can also be used as spermicides. These compositions can be used alone, with other known spermicides and with or incorporated into contraceptive devices such as condoms, sponges, vaginal inserts, contraceptive films, diaphragms, suppositories, contraceptive patches or sustained release devices. For use as spermicides, these compositions of the invention can be applied alone; with other microbicides; and with or incorporated into the contraceptive devices described above.
The composition can also be used in animals as disinfecting or antiseptic agent.
The microbicidal composition of the present invention consisting of an amino acid derivative that is modified with containing a C.sub.10-14 alkyl group at the N-terminus, particularly preferably N-cocoyl group is preferred. The amino acid residue bears a positive charge and the negative counter ion is organic molecule, preferably pyrrolidone salts. There are no halogen elements, such as chloride or bromide, in the structure of the amino acid compound. One general example is an amino acid ethyl ester acylated at the N-terminus with a coconut oil fatty acid residue, and having DL-pyrrolidone carboxylic acid added thereto.
The N-cocoyl amino acid pyrrolidone salts represented by the formula:
Wherein A is an amino acid residue and R is a C.sub.10-C.sub.14 fatty acid residue. In the structure, the amino acid residue could be from that of an arginine, lysine, and histidine; homolysine, or an unnatural amino acid residue bearing a positive charge, and other di or tri-peptide bearing a positive charge group.
For example, the N-cocoyl Arginine pyrrolidone salts is prepared with the following procedures:
Preparation solution A: Mix 1.2 equivalent N,N′-Dicyclohexylcarbodiimide (DCC) with 1 equivalent myristic acid (CH3(CH2)10COOH) in DMF and stir for 45 minute to 2 hours, Add in 1.2 equivalent N-Hydroxylsuccinimide (NHS) dissolved in CH2Cl2, keep stirring until precipitate forms, which is the undesired side product dicyclohexane urea. Filter the reaction mixture to remove the precipitate urea, and save the solution as solution A.
Preparation solution B: L-Arginine ethyl ester dihydrochloride and 1 equivalent of NaOH at 0 degree C. in DMF or CH2Cl2 for 30 minutes, and then add 1.3 equivalent of triethyl amine (Et3N). Mix to a homogeneous solution and save as solution B.
Preparation of the amino salt: Mix solution A and B together at room temperature for 2 hours. Remove most of the solvent by vacuum. Dissolve the product in methanol and add one equivalent of NaOH, and extract the product with a mixture of water and ether/EtOAc. Save and dry the organic phase under vacuum. Re-precipitate the product by using solvent hexane or water or methanol, or a mixture of methanol and water. Re-dissolve the product in methanol, and add one equivalent of DL-pyrrolidone carboxylic acid, and then remove the methanol under vacuum to obtain the final product.
In the formulation of the present invention the N-cocoyl amino acid pyrrolidone salts is contained usually in a proportion of from 0.003 to 3.0% (w/w).
The N-cocoyl amino acid pyrrolidone salts are soluble in water and highly safe with respect to acute toxicity (LD.sub.50). It has no irritation to skin and ophthalmic mucosa. Moreover, it has good biodegradable characteristic and can rapidly be decomposed in wastewaters.
In the microbicide compositions of the present invention a natural plant polysaccharides could be incorporated in the formula to enhance the antibacterial effect and protect the epithelium tissues. The plant polysaccharides may, for example, be Aloe Vera polysaccharides.
In the microbicide compositions of the present invention an ampholytic surfactant may be added and incorporated so long as it does not inhibit the effect obtained by the N-cocoyl amino acid pyrrolidone salts, in an amount which does not impair the antimicrobial effect. The ampholytic surfactant may, for example, be an amine oxide such as alkyl dimethyl amine oxide.
In the present invention, in addition to the specific active ingredient, one or more additive components conventionally used may further be incorporated in a conventional amount, as the case requires. The additive can be used including natural polysaccharides, natural plant acid and their salts, fragrance, colorant, flavor, plasticizer, stabilizing agent, emulsifier or moisturizer. Such an additive component is preferably one, which has a high safety for the human body, which is adequately compatible with active ingredients, and which has no influence over the effectiveness and stability of the composition.
In order to facilitate a further understanding of the invention, the following examples are presented primarily for the purposes of illustrating more specific details thereof. The invention is not to be deemed as limited thereby except as defined in the claims.
The liquid microbicide composition of the present invention is prepared by adding and incorporating the active ingredients in proper amounts into a proper aqueous medium in an optional order in a conventional method:
The gel microbicide composition of the present invention is prepared by the following procedures:
Aloe Vera:
Keep stirring until the mixture's temperature dropped to room temperature to obtain a colorless transparent gel.
An appropriated amount (3-5 ml) of gel was filled into a syringe. The filled syringe was sealed within an aluminum foil bag. At use, the user open the foil bag and insert the syringe to vagina or rectum to apply the material.
Table 1 listed 37 formulations. They are prepared following the above procedures to make liquid or gel preparations.
Aloe Vera powder
Aloe Vera
The testing results showed: Testing Sample diluted for 100 times and reacted with Ureaplasma Urealyticum suspension for 1 minute killed Ureaplasma Urealyticum.
The Killing Effect of Testing Sample on Neisseria Gonorrhoeae (Table 3)
The testing results showed: Testing Sample diluted for 100 times and reacted with Neisseria gonorrhoeae suspension for 1 minute, killed Neisseria gonorrhoeae.
Effects of the Microbicide on Trichomonas vaginalis
The Killing effect of testing sample to Trichomonas vaginalis (Table 4)
The testing results showed: Testing Sample diluted for 100 times and reacted with Trichomonas vaginalis suspension for 1 minute killed Trichomonas vaginalis.
HIV-1 titration was 107 TCID50.
The “drug removed control group” showed that the dilution could stop the effect of sample to the virus.
The Testing sample at 1:20 and 1:40 dilution reacted with HIV-1 for 1 min, titrated and cultured with MT4 cell, no cell abnormality were observed
The Testing sample at 1:80 dilution reacted with HIV-1 for 3 min., titrated and cultured with MT4 cell, no cell abnormality were observed.
The Testing sample at 1:160 dilution reacted with HIV-1 for 5 min., titrated and cultured with MT4 cell, no cell abnormality were observed.
Testing sample diluted at 1:20 (Example 1, formulation 15) and 1:40 solutions reacted with HIV-1 suspension for 1 min; diluted at 1:80 solution reacted with HIV-1 suspension for 3 min; diluted at 1:160 solution reacted with HIV-1 suspension for 5 min, all the kill rates were 100%.
The Minimum Bactericide Concentration (MBC) of Testing Sample to Candida albicans
Nutrient broth dilution method, Temperature 21±1° C.
To Candida albicans the MBC of testing sample is it's 16 times dilution (2 min).
Testing sample diluted at 1:20 (Example 1, formulation 7) and 1:40 solutions (Example 1, formulation 6) reacted with HIV-1 suspension for 1 minutes, the kill rate was 100%.
Testing sample diluted at 1:80 solution reacted with HIV-1 suspension for 3 min, the kill rate was 100%.
Treponema
Pallidum
Testing sample diluted at 1:10 (Example 1, formulation 10) deactivated Treponema Pallidum within 1 min.
Testing sample diluted at 1:20 (Example 1, formulation 7) deactivated Treponema Pallidum within 3 min.
Testing sample diluted at 1:40 (Example 1, formulation 6) deactivated Treponema Pallidum within 5 min.
Testing sample diluted at 1:80 (Example 1, formulation 1 1) deactivated Treponema Pallidum within 10 min.
The Effects of the Microbicide on Chlamydia trachomatis
The tests followed the “Standard” with consideration of the characteristics of Chlamydia trachomatis.
Killing Effects of Testing sample to Chlamydia trachomatis (Table 7)
Testing sample diluted at 1:50 (example 1, formulation 9) and reacted with Chlamydia trachomatis suspension for 1 minute killed Chlamydia trachomatis.
Testing strain: Ureaplasma Urealyticum: international standard strain; used 6th generation
Culture medium: High efficiency Ureaplasma culture medium;
Testing sample: Microbicide gel prepared as of Example 1, formulation 5.
2. Testing Methods and Standards: Technical Standard for Disinfection
(2002 Edition) Issued by “The Health Department of China”
The tests followed the “Standard” with consideration of the characteristics of Ureaplasma Urealyticum
The Killing Effect of Microbicide gel to Ureaplasma Urealyticum (Table 8)
The testing results showed: Testing sample diluted for 200 times (Example 1, formulation 4) and reacted with Ureaplasma Urealyticum suspension for I minute, killed Ureaplasma Urealyticum.
The Effects of the Microbicide on Neisseria gonorrhoeae
The tests followed the “Standard” with consideration of the characteristics of Neisseria gonorrhoeae
The Killing Effect of Microbicide gel to Ureaplasma Urealyticum (Table 9)
The testing results showed: Testing sample diluted for 100 times (example 1, formulation 8) and reacted with Neisseria gonorrhoeae suspension for 1 minute killed Neisseria gonorrhoeae.
The Effects of the Microbicide on Trichomonas vaginalis
The tests followed the “Standard” with consideration of the characteristics of Trichomonas vaginalis
The testing results showed: Testing sample diluted for 100 times (example 1, formulation 8) and reacted with Trichomonas vaginalis suspension for 1 minute, killed Trichomonas vaginalis.
The tests followed the “Standard” with consideration of the characteristics of Herpes Simplex
The Killing Effect of Microbicide gel to Herpes Simplex Virus (Table 11)
The testing results showed: Testing sample diluted for 20 times (Example 1, formulation 7) and reacted with Herpes Simplex Virus. Herpes Simplex Virus was killed with 20 times dilution of the sample in 1 minute.
The Effects of the Microbicide on Staphylococcus aureus
GB 15979-2002 Technical Standard for Testing Disinfection Product
“Procedures for testing bactericide effect”
The bactericide effect of testing sample to Staphylococcus aureus
At 20±1° C. temperature, three repeated tests showed: The testing sample reacted with Staphylococcus aureus for 1 minute the average bactericide rate was 95.70%. (table 12)
Testing sample (Example 1, formulation 5) reacted with Staphylococcus aureus for 1 minute the average bactericide rate was 95.70%.
The Effects of the Microbicide on Candida albicans
Temperature: 20±1° C.
The bactericide effect of Testing sample to Candida albicans ATCC10231 At 20±1° C. temperature, three repeated tests showed: Testing material reacted with Candida albicans ATCC10231 for 1 minutes, the average bactericide rate was 95.47%. (Table 13)
Testing sample reacted with Candida albicans for 1 minutes, the average bactericide rate was 95.47%. The Microbicidal Gel has bactericide effect to Candida albicans.
The Effects of the Microbicide on Eschetichia coli
The bactericide effect of Testing sample to Eschetichia coli:
At 20±1° C. temperature, three repeated tests showed: the sample gel reacted with Eschetichia coli for 1 minutes, the average bactericide rate was 99.97%. (Table. 14)
Testing sample reacted with Eschetichia coli for 1 minutes, the average bactericide rate was 99.97%.
Spermicidal Effects of the Microbicide Solution (in vitro).
The undiluted solution reacted with sperms for 30 seconds. The survival rate of sperms was zero. Table 15, 16 and 17:
The undiluted solution reacted with equal volume sperm for 30 sec, 1 min, 2 min, 3 min. The sperm survive rate was zero. The spermicidal rate of testing microbicide solution was 100% in 30 seconds (in vitro). Saline reacted with equal volume sperm, the sperm count and active rate was not influenced.
The spermicidal rate of testing sample was 100% in 30 seconds (in vitro).
Spermicidal Effects of the Microbicide Gel (in vitro)
The undiluted gel reacted with sperms for 30 seconds, the survive rate of sperms was zero;
The gel diluted at 1:1, 1:5 and reacted with equal volume sperm for 30 sec., 1 min., 2 min. and 3 min. The sperm survive rate was zero;
The gel diluted at 1:10 and reacted with equal volume sperm for 30 sec. The sperm survive rate was 10%;
The 1:10 gel dilution reacted with sperm for 1 min., 2 min. and 3 min. respectively the sperm survive rate was zero.
The 1:10 dilution deactivated all the sperms for 60 seconds. The survive test showed the sperm survive rate was zero.
The spermicidal rate of testing microbicide gel was 100% in 30 seconds (in vitro), the minimum effective dilution was 1:5.
The spermicidal rate of testing microbicidal gel was 100% in 60 seconds (in vitro), the minimum effective dilution was 1:10.
This application is based on, and claims benefit of, U.S. Provisional Application Ser. No. 61/001,936, filed on Nov. 7, 2007.
| Number | Date | Country | |
|---|---|---|---|
| 61001936 | Nov 2007 | US |
