I 1111111111111111 1111111111 111111111111111 IIIII IIIII 11111 111111111111111111 US010434110B2 c12) United States Patent (IO) Patent No.: US 10,434,110 B2 DeLuca et al. (45) Date of Patent: Oct. 8, 2019 (54) USE OF HOMOSALATE AND OCTYL Agranoff BW & Goldberg D (1974) Diet and the geographical SALICYLATE TO TREAT MULTIPLE distribution of multiple sclerosis. Lancet (London, England) SCLEROSIS 2(7888): 1061-1066. Becklund BR, Severson KS, Vang SV, & DeLuca HF (2010) UV (71) Applicant: Wisconsin Alumni Research radiation suppresses experimental autoimmune encephalomyelitis Foundation, Madison, WI (US) independent of vitamin D production. Proceedings of the National Academy of Sciences of the United States of America 107(14):6418- (72) Inventors: Hector F. DeLuca, Deerfield, WI (US); 6423. Steven Marling, Madison, WI (US); Compston A & Coles A (2002) Multiple sclerosis. Lancet (London, Lori A. Plum, Arena, WI (US); England) 359(9313):1221-1231. Yanping F. Wang, Madison, WI (US) Couteau C, Chauvet C, Paparis E, & Coiffard L (2012) UV filters, ingredients with a recognized anti-inflammatory effect. PloS one (73) Assignee: WISCONSIN ALUMNI RESEARCH 7(12):e46187. FOUNDATION, Madison, WI (US) Farah AE & Rosenberg F (1980) Potential therapeutic applications of aspirin and other cyclo-oxygenase inhibitors. British journal of ( *) Notice: Subject to any disclaimer, the term ofthis clinical pharmacology 10 Suppl 2:26ls-278s. Goldberg P (1974) Multiple sclerosis: vitamin D and calcium as patent is extended or adjusted under 35 environmental determinants of prevalence. International Journal of U.S.C. 154(b) by O days. Environmental Studies 6(1):19-27. MacLaughlin JA, Anderson RR, & Holick MF (1982) Spectral (21) Appl. No.: 15/643,988 character of sunlight modulates photosynthesis of previtamin D3 and its photoisomers in human skin. Science (New York, N.Y.) (22) Filed: Jul. 7, 2017 216(4549): 1001-1003. Marusic S, et al. (2008) Blockade of cytosolic phospholipase A2 (65) Prior Publication Data alpha prevents experimental autoimmune encephalomyelitis and US 2018/0015107 Al Jan. 18, 2018 diminishes development of Thl and Thl 7 responses. Journal of neuroimmunology 204 (1-2):29-37. Related U.S. Application Data Miller H, Newell DJ, & Ridley A (1961) Multiple sclerosis. Trials of maintenance treatment with prednisolone and soluble aspirin. (60) Provisional application No. 62/362,951, filed on Jul. Lancet (London, England) 1(7169):127-129. 15, 2016. Miyamoto K, et al. (2006) Selective COX-2 inhibitor celecoxib prevents experimental autoimmune encephalomyelitis through COX- (51) Int. Cl. 2-independent pathway. Brain a journal of neurology 129(Pt A61K 311618 (2006.01) 8):1984-1992. A61K 8137 (2006.01) Modi, Khushbu K et al., "Up-regulation of Ciliary Neurotrophic A61K 9/00 (2006.01) Factor in Astrocytes by Aspirin: Implications for Remyelination in A61Q 17104 (2006.01) Multiple Sclerosis," Journal of Biological Chemistry, vol. 288, No. A61K 31/60 (2006.01) 25, May 7, 2013, pp. 18533-18545. (52) U.S. Cl. Paulus HE & Whitehouse MW (1973) Nonsteroid anti­ CPC A61K 311618 (2013.01); A61K 8137 inflammatory agents. Annual review of pharmacology 13: 107-125. (2013.01); A61K 9/0014 (2013.01); A61K Rose JW, Hill KE, Watt HE, & Carlson NG (2004) Inflammatory 9/0019 (2013.01); A61K 9/0053 (2013.01); cell expression of cyclooxygenase-2 in the multiple sclerosis lesion. A61K 31/60 (2013.01); A61Q 17104 (2013.01); Journal of neuroimmunology 149(1-2):40-49. A61K 2800/91 (2013.01); A61K 2800/92 Tsau S, Emerson MR, Lynch SG, & LeVine SM (2015) Aspirin and (2013.01) multiple sclerosis. BMC medicine 13:153. Wang Y, et al. (2013) Suppression of experimental autoimmune ( 58) Field of Classification Search encephalomyelitis by 300-315nm ultraviolet light. Archives of None biochemistry and biophysics 536(1):81-86. See application file for complete search history. Wang Y, Marling SJ, Martino VM, Prahl JM, & Deluca HF (2016) The absence of 25-hydroxyvitamin D3-lalpha-hydroxylase poten­ (56) References Cited tiates the suppression of EAE in mice by ultraviolet light. The Journal of steroid biochemistry and molecular biology. FOREIGN PATENT DOCUMENTS * cited by examiner WO W02010/076731 * 7/2000 ......... A61K 31/4184 OTHER PUBLICATIONS Primary Examiner - Jake M Vu (74) Attorney, Agent, or Firm -Quarles & Brady LLP Orton et al (Association of UV radiation with multiple sclerosis prevalence and sex ratio in France. Neurology. Feb. 1, 2011; 76(5): 425-431).* (57) ABSTRACT Koriem et al (Multiple sclerosis: New insights and trends. Asian Pacific Journal of Tropical Biomedicine. vol. 6, Issue 5, May 2016, A method of treating an MS patient with homosalate, octyl pp. 429-440).* salicylate, or a combination is disclosed. International Search Report and Written Opinion issued in corre­ sponding International Application No.PCT/US2017 /041092, dated Aug. 30, 2017, 7 pages. 10 Claims, 12 Drawing Sheets U.S. Patent Oct. 8, 2019 Sheet 1 of 12 US 10,434,110 B2 1 3 5 7 9 11 13 15 17 19 21 23 25 27 Sunscreen Days Post Immunization Fig. 1 U.S. Patent Oct. 8, 2019 Sheet 2 of 12 US 10,434,110 B2 The Effect of Sunscreen Ingredients on EAE - Test 1 3.5 ~ Control ~ 8 3.0 VI ~ 2.5 "0 •.•.•.•-:=,•-•.•.• Dxybenzone .E 2.0 QI •.•.•·''·"-····· Avobenzone ~ 1.5 QI ~oxy+Avo i5 1.0 g:, 0.5 ~Sunscreen ~ l o.o .:.. s::,, s:: s:: s:: s:: s:: $;!' """. -~, ..... , ........ 1 3 5 7 9 11 13 15 17 19 21 23 25 27 ·-29 Days Post Immunization Fig. 2A 24.0 23.0 22.0 ~Control :§ 21.0 -~DMSO 3 20.0 ......,,,,, ....... Oxybenzone > -a ~ 19.0 ......., .. ,....... Avobenzone 18.0 - Oxy + Avo 17.0 ~Sunscreen 16.0 1 2 3 4 5 Weeks Post Immunization Fig. 2B U.S. Patent Oct. 8, 2019 Sheet 3 of 12 US 10,434,110 B2 The Effect of Sunscreen Ingredients on EAE - Test w~ -N~~w 2 6 Control ..... A ,X 3. 0 .. :❖• ••. ···,~•.,.•• .-·. .,,•• , .••••• ra ····•·:~~--·.·· Homosalate s~·····-·-=-:<"•. ...... •.·•-::~......... :~.......... ::.•-•.•·-... ❖-•··•·..:..::-·'"•.•·· --❖-----.°'::-....... :y. j2D ;· uc: 1 O ·1 .......,,;;:.•.•.• octyl .... ······ ;.<' •.. •.. ,:::- •..•..::,·······'"''······•·-:: •••.•.• ,,,;:"······='=····••.,,,,. ' . ... :::::·•··• .•.• .v·••:❖:•••:•:❖•-·•:::::•··•·::;.:-·•···•;::(" ~ o:o l::::: ....... s:ic~at~ *. *. * ·L·:-t··:·:::·:::i ~-· & . % . «:.:::::. ;_~ .·:::::·. 'k:W· ~:·w·:w -w. ·:·::: ..¾ .•~~.<-Homo 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 Days Post Immunization Fig. 3A Weights 25.0 24.0 23.0 ::§ 22.0 ...... +•-•.•. Negative Control 3 21.0 ......;:;:;.-.-.-. Homosa late ~ 20.0 ......,,,,, ....... octyl salicylate ~ 19.0 18.0 ·••·"····· Homo + Octi 17.0 ~sunscreen 16.0 1 2 2 4 5 Weeks Post Immunization Fig. 3B U.S. Patent Oct. 8, 2019 Sheet 4 of 12 US 10,434,110 B2 Fig. 4A 4.5 ! ■ Control 4 .0 !~NBUVB 3.5 i ~ SS+NBUVB ~ 3.0 0 65 2.5 ~ 2.0 <I) :;§: 1.5 1 0 · . TT TT TT L 0. 5 +❖~---w·1.JlL1 · · +.-0-,tA::A=' Q-¼;l,~~;¾nfl,#~h◊,~~/4-t<w&Y'{k:".!fy.:$-t%,<~~~-,¢y;~-0-,-$:-, 1 3 7 9 11 13 15 17 19 21 23 25 27 29 Days Post-Immunization Fig. 4B 25 20 5 ■ Control ❖ NBUVB *SS+NBUVB o+====;:::::::..-----,-----.----....-------.. 2 :3 4 5 Weeks Post-Immunization Fig. 4C 4.5 ■ Control 4 .0 4NBUVB 3.5 ~ss .§ SS+NBUVB ~ 3.0 0 bJ 2.5 ~ 2.0 <O ~ 1.5 1.0 0.5 0-l-®,%,<®-&:-;-%-,~~~M~~~-fi.:>.r~~r$.,-1:Nil~~~/4M~ 3 5 7 9 11 13 15 17 19 21 23 25 27 Days Post-Immunization U.S. Patent Oct. 8, 2019 Sheet 5 of 12 US 10,434,110 B2 3.5 Fig. 5A ■ Control 3.0 .& CotZ Face l· Banana Boat Kids 2.5 ~ Blue Lizard ~ 0 ,,,,, Kiss My Face !,) 2.0 (f) Y Coppertone (Spray) c ~ Hawaiian Tropic ~ i.5 ~ 1.0 0.5 O+M~~~;UU;~.,..,~~~~~~~ 3 5 7 9 11 13 15 17 19 21 23 25 27 29 Days Post-Immunization 3.5 Fig. 5B 3.0 2.5 ~ 0 u 2.0 (f) C ~ 1.5 ~ ■ Control 1.0 .& Darkness from -7 to 30 -,} Darkness from Oto 30 0.5 '? Darkness from 10 to 30 ~ Darkness from 20 to 30 3 5 7 9 11 13 15 17 19 21 23 25 27 29 Days Post-Immunization U.S. Patent Oct. 8, 2019 Sheet 6 of 12 US 10,434,110 B2 Fig. 6A Fig. 6B 4 30 ■ Control <I> Pretreatment 25 Immunization 3 ♦ x Onset § 20 :E ·a;"" 15 s °8 10 co 5 0 0+-------~----------~ 1 3 5 7 9 11 13 15 17 ·19 21 23 25 27 29 2 3 4 5 Days Post-Immunization Weeks Post¥lmmw1ization 4 .. 25 ■ Contrcl .,.ss 25 µL 50 µL 20 3 "'SS ♦ SS 100 µL § ~ t 15 (/J'·' 2 C. ~ .,"' ;;,, 10 2 '8 co 5 0 0+-------~----------~ 3 5 7 9 11 13 15 17 19 21 23 25 27 29 2 3 4 5 Days Po~t-lmmunization W..eks Post-Immunization Fig. 6C Fig. 6D U.S. Patent Oct. 8, 2019 Sheet 7 of 12 US 10,434,110 B2 Fig. 7A Fig. 7B 3.5 24 ■ Control ·····•: 3.0 ❖-DMSO ._ Ox)'ban2one 2.5 ,,,. Avobenzon e § 22 >); .-ii>-~-'&/;:::~;~1 6 ~ Oxy +Avo w 2.0 ~Sun-screen ff) £i ~ 20 .,,~ 1.5 2 t' 1.0 s 18 0.€, 0 ·&:("%y.$y:::!)'&·;·%'c (:::,1'::::"i-:;::i::$:,-::;-f:::-)'-»j•»("%f::•,-:-::-,.~~i•%'(%"f:::-{-::-i*-:~·(::;p,-, 16+-----,------,-----r-----,------, 3 5 7 9 11 r13 15 17 19 21 23 25 27 29 2 3 4 5 Days Post-Immunization \.,..,~leeks Post~ Immunization 3.5 24 ■ Control 3.0 -. Homosalate "Octisalate 22 2.5 -t>Homo+Octi :§ ~ •::: Sunscreen Spray! 0 :E w 2.0 OJ w ·;;; 20 C. 5 "'.,, 1.5 2 1.0 t Cll 18 0.5 0 :.:::y:::-<::::,:.::f'.::~1::::yZ.j:::.:c:::;:,.:~·-·/i:yt:l:•f:~:•1:~;..,;:;::{:::,.:::•1:~;--i=::~~%1 1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 2 3 4 5 Days Post-Immunization Weeks Post-Immunization Fig.