2, United States Patent oy oy 3) wo ay @ ws) (60) os) Foster et al. TREATMENT OF VULVAR PAIN Applicant: University of Rochester, Rochester, NY Us) David Foster, Rochester, NY (US): Megan I. Falsetta Wood, Rochester NY'(US): Richaed P. Phipps, Rochester, NY (US) Inventors Assignee: University of Rochester, Rochester, NY Us) Notice: Subject to any disclaimer, the term ofthis patent is extended or adjusted under 35 USC. 1846) by 247 days Appl. Now 16/590,973 Filed: Oct. 2, 2019 Prior Publication Data US 202010121617 AL Apr. 23, 2020 Related U.S. Application Data Provisional application No. 62/747,901, filed on Oct 19, 2018 Int. Cl, AGIK 31/121 (2006.01) GLP 15/02 (2006.01) A6IK 9700 (2005.01) oa cee AGIK 31/121 (2013.01), A61K 9/0014 (201301): ABIP 15702 (2018.01) Field of Classification Search cre AGIK 910014; A6IP 15/02 ‘Se application file for complete search history. 'US011400057B2 (10) Patent No.: (4s) Date of Patent: US 11,400,057 B2 Aug. 2, 2022 66) References Cited US. PATENT DOCUMENTS. $912,005 A 611999 Bockow et a aowdog7oea AL* 32014 Shan ALP 1100 2OIWOMIOTSL AL 42018 Sciavotino eta 2o1gm200395 AL 72018. Slavoline ea FOREIGN PATENT DOCUMENTS wo 2013170006 42 112013 (OTHER PUBLICATIONS Sobel eal, Femnle Seal Pain Disorders, Mar. 6, 2008, . 95-104 ‘Westelmann oa, ain, 20141559) 1696-1701 (ear: 2014)¢ Falbeti, M. e¢ aly "A Mch’Newded Mode forthe Prelnial Teding of New Valvodynia Therpies’s ural of Lower Genial Tract Disease 2017) vol. 21:4 Supplemental 1: 2 pas Dmiviovs, Nt al, “Resoline ReDI and 17(R)-RWDL Alleviate Signs of Inflammation in Rat Model Endomrones Feit and Sterly QO14); vol. 1024, pp. 119-1196 Maria, F, “Alpin Lipoic Acid Plus Omeas-3 Fatty Acids for Vesibuiodynia Associates with Phinfl Bladder Syndromes 1 Obstcr Gynaceol Can 2017); vol 39°3; pp. [31-137 (Continved) Primary Esaminer — Son Ming R Hii (74) Attorney, Agent, oF Firm — Fox Rothschild LLP on ABs The present invention relates to methods of treating female reproductive tract initation (Guch as pain and. pruritus) or/and inflammation, RACT Biopsies 3.3 mmUS 11,400,057 B2 Page 2 66) References Cited (OTHER PUBLICATIONS Serhan, CN. etal, “The Resolution Cade of Aste Inflammation Novel Pro-Resolving Lipid Mediator in Resoltions" Seminars in Immunology 2015), vl. 273; pp. 200-218 Falta. ML, etal, “A Review ofthe Available Clinical Therapies for Vulvodynia. Managsment and New Daa Implicating Pro- Inflammatory Mediators in Pain Elston”: BSG (2017) vo 142; pp, 210-218, Mysraym FA al, “Aatitnlanmatory Activity of Bass Oi ‘ant Development of & Microemulson System for Topical Dliv- ‘ey, Evidence-Based Complementary and Altomatve Medicine {G017), vol. 2017, 1 ps. Falela, ML. of aly "Specialized Pro-Resolviag Mediators: A Promising New Therpeitie Avene for Valvodymi Juma of [Lower Genital Tract Disease 2019), v.23, Suplemestal ‘Ramsden, CE. a, Diety linoleic avi-inlucad alterations in pro ant ant-neieetive lipad auacoi: Implications Tor iio- phic pain syadromes?; Molecular Pain (2016), vo 12; 14 pas. Hesselink et a, "New Topical Treatment of Vulvodynia Based fm the Paihogeneic Role ef Cross Talk Between Noviepors, Inmunocompetent Cel and Epiheti Cells Journal of Pa Research 2016), v0. 9; pp. 757-70 Tev-Sagie, Af al, "Recent Advances in Understanding Povoked ‘estbuodynia"; FIOOO Research (2016), 10 pas Sethan, CN. “Novel Pro-esolving Lipid Mediators in Inflam- ‘maton aye Leas for Resolution Physiology”, Nature (2014); vo. 510-7503: pp. 92-101, Falta, ML. eal ,“idemieation of Novel Mechanisms Involved In Generating Localized Vulvodynia Pain", Am. 3 Obstet Gynecol Gors) vol 13:1, 27 pes. Fost, D.C, “Site Specie Mesenchymal onto of infamnatory Pain to. Yeast Challenge in Valvodynia AMlited and PainsTree ‘ormen"; Pain (2013), vol. 186:3; pp. 386-396. * cited by examinerU.S. Patent Aug, 2, 2022 Sheet 1 of 10 US 11,400,057 B2 Vestibule FIG.1US 11,400,057 B2 Sheet 2 of 10 Aug, 2, 2022 U.S. Patent sioqeipeut Aroyeuueyuroid Woy uononpoud Joyeipawi ured-oud PReeuue ao ‘ul uoHONpes 4ysaje0i6 uo peseq Qd BANOS a jSOW JuiLUIA}Eq 1 uny az ‘Old Adj uewny oii Oy uled pue uled jo ese anyoayja SOLU dem eatectinen {pg/ml} 8 Vehicle Zymosan 10* 405 402 B 3 8 3 BE 3000: 3S | & & 20004 ul o a Zymosan - Vestibular Vulvar Vestibular Vulvar Controls (n=4) LPV Cases(n=5) C2 | @ Log of Pain Threshold) Pa |= Fitted Values @31 [£0 195% Confid, Interval gs £90 52 2s ey = 0 4000 2000 3000 4000 5000 PGE, Production from Cultured Vestibular Fibroblasts (pg/ml) FIGs. 3A, 3B, and 3CUS 11,400,057 B2 Sheet 4 of 10 Aug, 2, 2022 U.S. Patent Gy pur “Dp “ab “Vb “SDI 210149 oF & z oor 3 coo & é jooo'z 3. a RIA 3 wary? in z ooo 8 5 00 & ie & 1 ono 2 3 v TRUEST" peesUS 11,400,057 B2 Sheet 5 of 10 Aug, 2, 2022 U.S. Patent sol {juyBd o4) de-t (wu) Luisereyy CO} Ob baie SION 3 000°) “y x oovoz 3 o0o'0e § * coos & o00'06 3 oo0'o SSUS 11,400,057 B2 Sheet 6 of 10 Aug, 2, 2022 U.S. Patent D9 pur “a9 “V9 "SOLA ah A AOR OL+ e+ a+ b+ 0 PSsey sos Tyson) SUL Aasg UOA 21U0N}99/9 Buisy) + sBAIB8O papuyig + Laseyg wioy paztuwopuey + Li bhid? paweyuy = uonsalu-aig WWUS 11,400,057 B2 Sheet 7 of 10 Aug, 2, 2022 U.S. Patent old (sem y) eseUd (Sy@OM Q) eseud WOUWIEAL sousisisiog uoqonpuy ye fb ———— 95 PE CTO heen net eednetgs QQ mL Ae (010) swiei6) ppoyseiyy wegUS 11,400,057 B2 Sheet 8 of 10 Aug, 2, 2022 U.S. Patent 16eq (uy6d) "39g eBeaey jeu! (syeem p) Z eseud jueuie $) Z e8e8Ud ee 0 000‘. 00002. oo0'oe. ot tepntnntny Of BB 2394 [eUlbe A, flee PIOYSAIYL UIE =e (sem g) | eseyd UORONpU| Sp € 2@+4 0 “2 2 & a oa oi {so10) swei6) pjoyseayL ued s 3 ae 3US 11,400,057 B2 Sheet 9 of 10 Aug, 2, 2022 U.S. Patent 6 Old SINdS PeAuep-WHG (panuep (peruep ~yd3)-v¥) SHOGHIG ‘any "ad TXT Luise ‘ag = Fan “SZL'SOL ‘Gk Oxo-g PE pee Rain, jeulayeg ae AT ill ainqnsen ese AdT BAINA [BUIAKA JOU — BINGNSAA [ONUOD [7] ° 002 3 = ‘009 008U.S. Patent Aug. 2, 2022 Sheet 10 of 10 US 11,400,057 B2 < S 2 2 & S N [+0 s 1]? 3S S 3 og 3 8 s Ss S S E 2 2 8 © (uySd) -nu09 *g9g F “I + G e age ° =| 2 8 S 4 2 g Ga = = £| a = é s a eens é (6) e104 pjoysaiyy OQ (4A) Aerg von jenuey + x $ = Week 0 o + Oo Ne (6) e404 pyoysasyy (4AM) Aary uo penuew A Induction Phase 4US 11,400,057 B2 1 TREATMENT OF VULVAR PAIN GOVERNMENT INTERESTS ‘This invention was made with goverament support under HIDOG9313 awarded by National Institutes of Health. The government bas cetsin rights inthe invention. FIELD OF THE INVENTION This invention relates to treating female reproductive tract imitation orand inflammation. BACKGROUND OF THE INVENTION Invitation, such as psin and pruritus, ftom the female reproductive tact is a Siuifican clinical problem for whieh thre are few effective therapies, Vulvar pain or persistent vulvar pain can be caused by spoifie disorders soch as infectious (es. ecurenteandid- ais, herpes, trichomonisis, ete) inflammatory (e ichen Selerosus, lichen plans, immanobolous disorders, ete); nenplostic (eg Paget disease, squamous cell carinoms, ‘ce nourlogic (es. postherpetie newly, nerve com: pression or igure, neurons, et) wauna female genic ta cating, obstetrical, et): atogeic (ex. postopentive, ehemoteapy, radiation, et); hormonal deficiencies (ea gento-urinry syndrome of menopatss, vulvo-vaginal ao Phy lactation amenorrhea, et) Th adlition, vulvar pin can be idiopathic which is case siti as vulvodynia."™ Valvodsnia 2 vu eronie pain fof at least 3 months of duration, without clear identifiable ‘ause, and may have potential associated factors. Vulvedy= nia alloca the vulva, the external female genital ona. “This includes the labia clitoris, and vaginal opening. Pin is the most nouble sympiom of vulvedyai, an canbe care scerza asa burning, stinging tation o sarp pein that ‘cts the vulva and eniancet the Vagina, This pai may he gonorlizd andior localized (vesibuledynia,citorody na, hemivolvodynia), constant, inlrmiten oF provoked {happen only when the vulva is ouched). The mast eommon subse of vulvodyia ib localized provoked vulvodyaia (LEV) which is characterized by acute and lasting pai in response to light (ouching of the vulvar vestibule (area ‘mmeditely suounding the vaginal opening) act as many ay 1 in 3 women within their iftime and causes Significant poyshologicel distress and sexsal dysfnetion Glariow, BIT: Kunitz, CG: Nawyen, RH Rydell, 8 Tomer, RM; Maclehose, RT. Am J Obstet Gyeso! 2014, vol 210, pp. 40 el-8). Therefore, LPV is ‘women's heath issue Ti the most sever cases, women may have both a pain ‘cause by # spocie disorder and wulvodynia, “Teatment of itation, such as vulvar pain and prarits, may involve @ numberof ciffeent measures” However, al ‘currently avilable therapies only manage pain and psycho- logical distress but none is universally effective to date and the evidence to support their ellectivenes is often poor 0 o 2 1 is really remarkable that current treatments have not addressed the underlying biological causes of disease and, therefore, an approach in this regard could improve even restore the patients quality of lie “The compounds mentioned in the present invention have ben disclosed in WO2010033809 and WO2013170006 for the treatment of differet inflammatory diseases but-not specially for the treatment of female reproductive tact irritation, SUMMARY OF INVENTION This invention relates to treating female reproductive tract fo, such a pain and pruntis, or/and inflammation, “Accordingly. in one aspect, the invention provides a method of reducing. preventing, or treating lower genital ‘woot ination in a subject comprising administering to a subject in need an effective amount of at least one compound having the Forma (Ds wherein ‘wherein isa double bond withthe Z or P confign- withthe Z or F configuration; ‘wherein the carbon at C7 and C14 are, independently, either R or Si wherein R,, is selected from hydrogen, (C1-C6) alkyl alyeeryl (C3-C8)eyetoalkyl, cyelohexyl, (C4-CH) eyeloal- Jelalkyl, (CS-C15) aryl, (C6-C16) arylalkyl, 2-6 membered heteroalkyl, 3-8 membered eyeloheteroalkyl, 4-11 mem- bred cycloheteroalkylalkyl, (CS-C14) heteroaryl and 6-21 membered heteroarylalkyl: ‘wherein each of Ro and Ry: is independently a hydroxyl soup of hydrogen stom, with the proviso that at least one Of Ry and Ry. is 3 hydroxyl group: wherein R,, if present, is selected from hydrogen, (Cl- 6) alkyl, (C3-C8) cycloalkyl, eyelohexyl, (C4.CH1) cycloalkylalkyl, (C5-C15) aryl, (C6-C16) arylaiy, and 26 srembered heteroalkyl; hercin Ry is selected from hydrogen, (C1-C6) aly (C3-C8) cycloalkyl, (CACM) cycloalkylalkyl, (C5-C15} aryl, (C6-C16) arylalkyl, and 2-6 membered heteroalkyl: ‘or a pharmaceutically acceptable salt thereof; and ‘optionally, « pharmaceutically acceptable carer In one embodiment, compounds of Fonnula (1) are inthe teglyceride,diglyeeride, and/or monoglyeeride form. Tn one embodiment, the ower genital tract irritation is genital tract pain. Tn a particular embodiment, the genital tact pain is iiopathie. In a more panicular embodiment, the idiopathic lower anita tract pain is vulvodynia Even more particulary, the vulvodynia is localized pro- voked vulvodynia (LPVUS 11,400,057 B2 3 Ina preferred embodiment, compounds of Formula (Dare Macesin-l, 78-Maresin-1, 14-methyl-Maresia 1, 7-methyl- Maresin I, 7,14-dimethyl-Maresin 1, and 14-hydroxy-doco- sahexaenoie acid (14-HDIA) and the corresponding accept- able pharmaceutical salt or esters thereof, The method can further comprise administering at leat ‘one compound of Formula (1) with one or more therapeutic agents (eg. one or more specialized pro-resolving media- tors (SPM), one or more SPM precursors, an anti-mierobial agent andor an antiviral agent) tothe subject, ‘The details of one or more embodiments of the invention are set forth in the description below. Other features, abjec- tives, and advantages of the invention will be appareat from the description and from the claims, BRIBE DESCRIPTION OF THE DRAWINGS IG. 1s adingram showing testes ofntense pin al he vestibule of EPV pation ae in close proximity 10 pon Pain sites oft extemal vulva and rated biopsy sts. IGS. 24 and 28 ae ast of diagrams showing proce hrs foe (A vetting the ably fo rie prin matey at pospain, menor preston form. primary fhmar cellin ann vito LPV model an (B) valtng the ‘cacy of compos in alevating pain wing pretinical mouse model of LPV." IGS. 34, 38 and 3C ara et of iagrans showing tht inflammatory mediator production is elevated in vestibular cll fom PV patent compared io vulvar cells or contol subjects. Panel A TL reeset in response to dseretsng fdowes of lve © licon. sp-008 vehicle vs. dose of © albicans (for vestibular cells only), **p<0.05 vestibular vs. {var ells fora prtular dove, ANOVA, 2. Vestibular ‘alls show a song reponse, while vulvar cells show no Significant response ton dose up fo 1000 times wean Pane B: Cultured fibroblast were stimulated with vehicle for Zsmosan (100 yim) for 24 hour, then modin were analyzed for PGE." Zymosan induced a significant incre in PGE, over conespnting vehicle treatment Vexibolar fibroblasts fom LPV case patients produced mnore PG, compared to vestboarfiolats fom conta fobjets Nisine/-SEM, ANOVA "pe 05, Panel Seater plot of fitoblast PGE, production plotted agains fog ans= Formation of mcoeitaneots pain threshold performed tefore time sampling, fom ideal anatomical ses Conta Tine: oid re) represents fied valves of ineat regression delinited hy 9% confidence interval (se ‘td ng) 1-2 58, 7-004, IGS, 4, 43, aC and 4D are a set of diagrams showing shat PGE, and IL-6 proton freed by compound of Forma (I). Patent vestbolar or vulvar Aboblt were pretest for 10 hours with Marein land eprMaresin Tat £SaNt concentration. then atvated wih (Panel A 10 ei) or bradykinin (Panel Bs 100 nM) fr 48 he Coors mia wore collected an analy for PGE (Panel) ot 1-6 (Panel B) content, Patent fibroblasts were also tie ated fist with IE foe 30 min then tested with $M Miresin I for 18 hour, alowed by a booster dose for 6 Cire media were collected and analyzed for PGE, (Panel ‘Chorlt6 Panel D) content Means/-SEM of 1-3, ANOVA *p=0.05 vs. activation only (no compounds of Formula (1) (Eq), These rents were consistent fr sever ona "PY pion sins FIG. Sia diagram showing that eompounds of Formula (0) Gy inkibit PGE, prvntion rom mene vv sue Mosse wlvar tse (fsa ponch biosin) wa collected and bisected hen pretretdin cute medium wither 0 o 4 ‘Mares | at indicated concentrations for 18 hours, Followed by an additional 18 hours stimulation with IL-1 10 pei) Culture medium was collected and analyzed for PGE, content, Meant/~SEM; 1=3 replicate cultures. ANOVA * 0.05 ¥s, vehicle FIGS. 6A, 6B and 6C are a set of photographs and iagrams showing pain esting for in vivo mouse vulvodynia ‘model. Panel A: After Zymosan injection, inflammation and redness become apparent, Arrow indicates injection Panel B: Image shoving how an electronie von Frey hai (Mousemet is used to apply fore to the mouse vulva, Panel C: Schematic of in vivo mouse model to establish then resolve vulvar allodynia, IG. 7 is @ diagram showing representative pain profile for a CS7BLI6 mouse. This boxplot series shows median ‘threshold values fora representative mouse over the indvc- ‘ion, persistence, and iretment phases. Measures collected teach week were tightly distributed, and thresholds were redineed 33% an! maintained until reatment with Maresin 1. when values rerned to baseline. FIG. 8 is 9 diagram showing that PGE: in vaginal lavage is associated with pain threshold. After induction of alloy tia for up to 6 weeks (indicated by reduced pain threshold, re line), Maresin 1 was applied to CS7BL/6 mouse vulvas ‘o reduce pain and inerease pain thresholl. PGE, content in ‘vaginal lovage samples (blue line) rapidly increases during the induction phase, indicating pain-associated inflamma ‘ion waning with treatment. SEM shown, n-8. FIG. 9 isa diagram showing compounds that are produced by vulvar fibroblasts, Fibroblasts were cultured for 48 h with IL-1B (10 pa/m), then culture media were collected and frozen immediately on dry ice under argon gas for targeted Jipidomie analysis. The predominating compounds detected were derived from DHA. FIGS, 104, 108 and 10 area sot of diagrams showing ‘manual von Frey assessment of pain threshold that denotes in threshold with treatment. Decreased thresh wan-teated mice after 4 weeks of injection reflects increased painsensitivty, Meane/-SEM, 1-8 saline, n-12 Zymossn, ANOVA #p-0 05 (Panel A). Ther peutic treatment after the induction phase increased pain thresholds. Means/-SEM, -7, p>0.05 (Panel B). Vul- vovaginal lavage Avid was analyzed for PGE2 content (Panel C). Mice receiving Zymosan had elevated PGF? in their lavage fd versus mice receiving saline injection Furermore, treated mice with allodynia had reduced PGE2. Mean/-SEM, n-7, ANOVA *p0.05, In te figures, F (1) means compounds of Formula (D. DETAILED DESCRIPTION OF THE INVENTION ‘This invention is based, atleast in part, on unexpected iscoveries that fibroblasts isolated and cultured from sites of pain in LPV patients produce very high levels of pro- Inflammatory an pean mesos compar pn ‘As disclosed herein, it was discovered that the vulvar vestibule expresses a unique inflammatory profile involving the elevated production of pro-pain and pro-inflammatory mediators, eg. prostaglandin E (PGE) and interleukin-6 ((L-6) by fibroblast stains isolated from the vestibule site (FIG. 1, “Vestbule”), Furthermore, elevated pro-inflamma- tory mediator release eoreltes with pain profiles in women. ‘Therefore, effective therapeutics for vulvodynia would ide- ally reduce pro-inflammatory signaling, while preserving the fatural ability of these cells to respond to harmful pro-US 11,400,057 B2 5 ‘flammatory stimuli. The investigation deseribed herein has ‘entified mechanisms by which hypersensitivity to certain inflammatory stimuli leads to heightened pain, Specifically, it was demonstrated that pain in LPV patients is directly ccorelated with the production of provinflammatory and pro-pain meditors from fibroblasts culture from biopsies ‘of painful sites (FIG. 1, “Vestibule”, As describe herein, fibroblasts producing high levels of pro-pain and pro-inflammatory mediators can. be isolated ‘rom patients at sites with intense, quantifiable pain. AS they abundantly produce pro-pain mediators and maintain their relevant phenotypes in culture, the primary vestibular fibro- blasts are valuable in modeling LPV and were used sue- ‘cessfully here to identify new therapeutic agents that can Be used 1o resolve atypical inflammatory mediator production in LPY patients that leads t0 regional pai This invention addresses the treatment of female repro- ‘ductive tract iertation, such as pain and pruritus. Specifi- cally, this invention provides a treatment of vulvar pain oF vulvar persistent puis associated to a specific disorder such 1 inflammation or/and idiopathic pain, and more paricu- larly to vulvodynia, whose etiology is unclear and where no effective medical therapy fas been developed In particular, the compounds represented by Formula (1) have resulted to be effective against lower genital tract invitation, such as vulvar pain, and more particularly t vulvodynia, among others. Accordingly, in one aspect, the invention provides a method of reducing, preventing, oF treating lower genital teactinitaton in a subject comprising administering to 2 subject in aced an effective amount ofa least ane compound having the Formala (Ds wherein — is @ double bond with the Z of FE coatigus ration; ‘wherein ‘with the Z or B configuration; ‘whore the carbon at C7 and C14 are, independently, citer R or S: wherein R,, is selected from hydrogen, (C1-C6) alkyl, ulyeeryl, (C3:C8) eveloalkyl, eyelohexyl, (C4-CI) eyeloal- Kylalkyl (C5-C15) aryl, (C6-C16) arylalkyl 2-6 membered heteroalkyl, 3-8 membered eycloheteroalkyl, 4-11 mem- bred eyeloheteroalkyalkyl, (C5-C14) heteroary] and 6-21 membered heteroarylalkyi: ‘wherein each of Ry and Rs independently a hydroxyl rou or a hydrogen atom, with the proviso that a least one FR, and Ry is a hydroxyl group: wherein R,. if present, is selected from hydrogen, (Cl C6) alkyl, (C3-C8)_eyeloallyl, cyclohexyl, (C4-C11) ‘eyeloalkyllkyl, (C5-C15) aryl, (C6-C16) arylalkyl, and 2-6 membered heteroalkyl; wherein Ry, is selected from hydrogen, (C1-C6) alkyl (C3-C8) cycloalkyl, (C4HCT) cycloalkylalkyl, (C5-C15) ay, (C6-C16) arylalkyl, and 2-6 membered heteroalky: 0 o 6 or a pharmaceutically acceptable sat thereof; and ‘optionally, « pharmaceutically acceptable carer In one embodiment, compounds of Formula (1) are inthe tiglyceride, diglyeeride and/or monoglyeeride form. Tn other embodiment, R, is methyl or etl ‘The compounds of Formula (I) are SPMs or SPM Prec In some embodiments, examples of compounds of For- mula (D inetude, but are not limited to: Maresin 1 (Malt; 7R, 4S clihydeoxy- penitl tract pain is vulvodynia. ‘Vulvodynia is the most common cause of longstanding ‘dyspareunia (painful sexusl intercourse) in premenopsussl ‘women, characterized by walvar pain of at least 3 months duration, withomt clear identifiable cause, which may have potential associated factors. The most common subtype of Vulvodynia is LPV characterized by pain t0 Tight touch limited to the vulvar vestibule surrounding. the vaginal ‘opening. In women with LPV, chronic vestibular pain is crippling, impacting every aspect of life and exacerbating ‘comorbidities, such as fibromyalgia and painful bladder, The ‘devastating impact of LPV includes sexual dysfunction, ingerility, depression, and even suicide, ‘Therelore, even more particularly the vulvodyni ined provoked wulviodynia (LPV), “The compounds of Formula (I) of this invention resolve ‘nlammation and pain without impsiting normal host defense. The resolution of inflammation and pain, once thought 1 be a passive process during whieh pro-inflam- matory signaling tapers of, is now known to be an votive process mediated by the compounds of Formula (I). The ‘compounds of Formula (1 actively reduce pro-inflammatory signaling, promote bacterial clearance, reduce pai, and ‘accelerate wound healing. Compounds of Formula (Dare not traditional ant-inflammatory agents and are not immu- osuppressive: they do aot allt the body's ability to sense and respond to infection or injury ‘As disclosed herein, compounds of Formula (1) ean be ‘ideal therapeutic agents for vulvodynia, as they foster ‘wound healing, promote bacterial clearance, and reduce pain and pro-inflammatory signaling. Although compounds of Forma I) have not been clinically tested as in ulvodyaia therapy, evidence presented here supports that these com- pounds’ are elfcacious i redvcing pain-provoking.pro- inflammatory mediator production and in tur, redace LPV- associated pain in vivo. Th one embodiment, this invention relates to using com= pounds of Formula (}) to promote bacterial clearance and reduce pain, and accelerate wound healing. Using an in vitro ‘model described herein, inventors identified compounds of Formula (1) highly effective in reducing IL-6 and PGE; production in cells when administered prior o inflanmatory islocal- o 8 stimulation, Furthermore, Maresin-l is highly effective reducing IL-6 and PGE, i aleady activated cells, suggest- ing his compound is effective throughout the entire disease process. The results described herein suggest that com pounds of Formula (I) are effective idiopathic, an inflam- ‘matory pain atsociated with a genital tract inflammatory condition, such as lichen planus, lichen scleosus, desqua- ‘mative inllammatory vaginitis, and trophic vulvovaginitis associated with breast cancer and vulvodynia therapies ‘As also disclosed herein, a robust and reproducible mouse del of LPV was developed to assess therapeutic inter vention apainst vulvar pain (he frst of its kind). The model couples real-time provin lummatory mediator quantification ‘with mechanical pain testing via an electronic von Frey to ‘monitor pain and inflammation over time. Inventors were able to establish stable allodynia in miee, lasting more than several montis. During allodynia induction, it was found ‘that pain thresholds decreased, while_ pro-inflammatory mediator Hevels (eg, PGE,) increased within collected vulvovagioal Mus, consistent our in vito findings. Inven- tors then trated mice daily with topical Maresin-1, which increased pain thresholds, while suppressing PGE, levels. The in vitro and in vivo findings disclosed herein suggest that topical application of compounds of Formula (1) ean reduce Vulvar pain and inflammation and Would repesent aa ‘deal therapy for vulvodynia, Tn one embodinent, one or more compounds of Form ())can be formulated with other SPMs andor SPM prec sors not included in Formula (1). Such SPMs and/or SPM precursors are described in WO2013170006 Ina particular embodiment, said other SPMs and/or SPM prccursirs not included in Forma (I) are selected fom: TS-HDHA (17S-hydroxy-dlovost-42,72,102.132.15E, 197Z-hexacnoie acid), TTRAIDHA —(7R¢hydroxy-dloo0se-47,72,102.192,15E, 197-hexaenoie acid), 18 S-HEPE (I8S-hydeoxy-cioosa-57,87,117,142,16E-pen- tacnnic acid) ISR-HEPE. (ISR hydroxy -eieoss-57,87,11Z, tacnoic acid) ‘oF a pharmaceutically acceptable salt or ester thereof: In other particular embodiment, the compound of For: ‘ula (1) is H-HDHA and the SPMs andlor SPM precursor ‘ot included in Formula (D) are 17-HDHA und 18-HEPE: ‘or a pharmaceutically acceptable salt or ester thereof 'As used herein “SPMs and/or SPM precursors” reers to specialized pro-resolvingmedistors and/or precursors thereof: SPM or SPM precursor is lipid-derived compound for substance that promotes the resolution of inflammation, eg, it can reduce one sign or symptom of inflammation in ‘cll or organism, 'SPMs represent a clas of pro-esolving, ant-poin and ant-inlamimatory lipids naturally derived from omega 3 and ‘omega 6 fatty acid that belp healing without compromising the body’s ability to defend against inflammatory insults (ex, infection or injury)? SPMs area genus with several families of potent endogenous bioactive products derived {rom precursors essential fatty acids FPA, DIA, arachidonic acid (ARA) and Docosapentsenoie acid (DPA) that are biosynthesized by positional and stereospevfie incorpor- ‘ion of one, two or three molecules of molecular oxygen into a polyunsaturated fatty acid (PUFA) using FPA, DFA, ALA fand DPA as substrates into a catalyzed reaction involving {iy acid Tipoxygenases, cyclooxygenase type-2, when sd by aspirin, and several cytochrome P480 oxi- 16-pen-US 11,400,057 B2 9 As used in this invention, SPM relates ta PUEAderived ‘enzymatically-oxygenated derivative that has potent anti- Jnflammatory aad resoluion-aetvating setvity and tht aes as endogenous regulator of the inflammatory response bring an inflamed tissue back towards its non-inlamed and healthy state. SPMs act as endogenous roceptor ligands oF allosteric modulators fo potently activate celular responses that conceitedly activate antiinflammatory actions and ‘expedite, stimulate, and tigger resolution of inflammation, “The term "SPM precursor” refers (o an enzymatically oxy nated derivative of a PUEA that requires an additional ‘enzymatic reaction to conver iC to a SPM. A SPM precursor ‘sa more proximate substrate forthe endogenous formation ‘of an SPM than the corresponding PUPA subsinite itself The SPMSs include several families of mediators, lipoxins, resolvins (eg. the F and D series), protectins and maresns Examples of SPM include Resolvin ED (RVEI; $8,12.18- telhydroxy-cicosa-67,8E,108,147,16E-pentaenoic acid) 18S-Resolvin ET (I8S:RVET;_58,12R,18Sstriydroxy-¢ osu-67.8E,10E,147,16E-pentaenoic acid), 20-hydroxy- VBI (58,1 2R.18R, 30-totrdhydoxy-cicosa-67, 8B. 108,147, 16F-pentaenoic acid), Resolvin F2 (RVE2; S8,18- dihydrony-cicosa-GF.87,117,147,16E-pentoenoic acid, Resolvin E3 (RVE3; 17.18R-dihyroxy-eicoss-S7,87.11Z. I3E1SE-pentaenoie acid), 185-Resolvin ES (18S-RVE3, 17,188-dihydroxy-vieose-S7,82,112,13E,15E-pentaenoie seid), 17,1 8-epoxy-eicoss-5Z,8Z,11Z,13E15E-pentaenoic ‘cid, Lipoxin Ay (LXAs: 55,6R,15S-teihydeoxy-eicosa-7E, DEWIIZ,I3E,172-pentaenoie acid), 1Sepi-Lipoxin AS (LXAS; 58,6R,LSReteydoxy-eieosa-7E9E 117,138, 17Z- Pentaenoic acid), Maresin 1 (MaRI; 78,148 hydroxy PAIQZ 162,197 -bexaenoie acid), 7S-Ma- 78,1 4S-cihydrony-dooosa-4Z.8E-10F, 12Z,167,19/-hexsenoie’ acid), 7S,148-JiEIDHA (7S.148- dihydroxy docoss-87.8F,107.12E,167,17-hexaesoie acid), Proveein D1 (PDI; 10R,17S-dihydroxy-docosa-4Z, TEAIF 1 3F,152,192-hexaenoie acid), 108,17S-HDHA (108,178 dihydeoxy-docosa-47,77,11F 132,15E197- bexaenoic acid), 48.218-aiFIDILA (148.218 -ihydroxy-ao- ‘cos-47,7Z,107,12E,167,197-hexaenois acid), 148,218 INDHA (48,218 -dinydroxy docosa-47, 72,102,128 162, 19Z-hexsenoic acid), T4R2ISFDHA (4R21S- hydroxy locose-47.7Z,107.13E.167,197-hexaesoic acid), I4R2IR-GHDHA (14R.21R-70% LPV patents** Repeated vul- Vovaginal infection in miee indvees vulvar allodynia and regional. hyperinnervation, simulating vulvodynia find- ings * Recent studies" have shown that LPV is associ- ated with inflammatory dysregulation, despite the fact LPV ‘does not present as a classical inflammatory disease. The ‘cardinal signe of inflammation are act pronounced or are vaguely present in both heathy and LPVallliced women, 0 although the is ‘istinctively different in LPV versus healthy patients.” Tn addition, it was found that there was a site-specific response to live yeast infection, whereby fibroblasts from sites of pain within the vulvar vestibule are inherently sensitive to yeast yeast products and produce elevated levels ‘of pro-psin/pro-inlammatory mediators compsred to fibro- blasts from non-painful sites of the extemal vulva (FIG. 3B). Funtnermore this response appeared to be an exacerbation of 4 normal inflammatory response, as fibroblasts fiom the vestibule of healthy women belt reduced. response to Zymosan (yeast cell wall product; FIG. 3B). More importanlly, there was a strong connection between LPV’ pain and inflammation: pin ia LPV patents was directly correlated with the prodition of peo-nflam- ‘matory and pro-pain mediators by fibroblasts cultured from biopsies of painful sites, when exposed to live yeast (FIG. 30). Example 2 In this example, assays were carried out to investigate the ‘of compounds of Formula (I) to reduce pro-inflam- matory and pro-pain mediator production from primary ‘human cells inthe in vitro LPV model desribed in Example 1 The inventors wsed one of wo treatment regimens proven effective in vulvar fibroblasts and other cells at low inane ‘molar concentrations (1-100 nM): 1) overnight pre-reat ‘meat, followed by another treatment 30 min prior to stimu lation with pro-inflammatory stiauli for 48 br with a third dose ofa compound of Formula (I) at 24 hr post-challenge, fr 2) posttreatment a compound of Formula (I ater 2 30 ‘min pre-treatment with inflammatory stimuli, flowed by booster dose 18 hr later, Both treatment regimens ae of intrest, as compounds of Formula (I) ae active throughout the inflammatory process.” Even the compound of Formula () adasinistered after LPV onset are likely t preveat the worsening or spread of LPV pain ‘Te dats show that compounds of Formula (1) were highly effective in reducing pro-inflammatory mediators linked t0 pin in human vlvar Sbroblass treated with relevant pro- inlammstory stimuli using a pre-treatment strategy. Maresin 1 and epi-Maresin 1 significantly redveed prostaglandin Es (PGE FIG. 4A) and interleukin (IL-6: FIG. 4B) prodne- sion by both vestibular and extemal vulvar fibeoblass ‘Maresin 1 was also tested using the post-treatment regimen and. found both significantly reduced Il.-6 and PGE, levels under this strategy (FIGS. 4C and 4D). ‘Next, compounss of Formos () are effective in reducing IL-6 and PGE, production (eg, Maresin 1) are investigated in similar seraen. Live C. albicans yeast, Zymosan, brady Kinin, and IL-1P are used as different classes of inflamma- tory activators, which have been shown to induce the production of pro-inflammatory mediators in vulvar fbro- biasts°**:' Prosinflammatory mediator levels are mea- sured using ELISAs, ELAS, and Lumines assays. Com: pounds of Formula (I) are elective in ducing, more than ‘one pro-inflammatory mediator in at last 2 tests move on for futher testing using a preclinical mouse model. Con- agruent with the above supporting results showing several ‘compounds of Formula (I) were effetive in redoing pro- inflammatory mediator production, inventors expect to iden- {ify several adlitional compounds that are highly effective ‘ompounds of Formula (I) meeting criteria for furer