2) United States Patent Witham et al ($4) METHODS AND COMPOSITIONS OF STABLE PHENYLEPHRINE FORMULATIONS (71) Applicant: Paragon BioTeek, Inc, Portland, OR ws) (72) Inveators: Patrick HL Witham, Pugene, OR ( Sailaja Machiraju, Beaverton, OR (OS); Lauren Mackensle-Clark Bluett, Milwaukie, OR (US) (73) Assignee: Paragon Bioteck, In. Portland, OR (Us) (4) Notice: Suijct to any disclaimer, the tem of this patent is extended of adjusted under 35 USC. 1540) by O days (21) Appl. Nos 14/080,771 (22) Filed: Now. 14,2013 6) Inc. AGIK 31/137 (2006.01) (2) US.CL cP AGIK 31/137 (2013.01) usPc ‘514/658 (58) Field of Classification Search None See application file for complete search history. 66) References Cited USS. PATENT DOCUMENTS 420,600 8 * OTHER PUBLICATIONS 41981, Valle Akon, le, package inert for phenylephrine hydrochloride solo tion drops a hp dailyrped nlmaih gow dalymed Took. fin eid SeS1Sb-b500-4 77-9455 {Ele AYA, revised S 'US008859623B1 (10) Patent No. 4s) Date of Patent: US 8,859,623 BI Oct. 14, 2014 Bown, et aly “Activities of sereviomers of entrenosepors LShibin, ta. “The Stability of Phenylephrine Past The Rate of Degradation of the Amino Group Atmeimilfrschuns. Ape 1909:196.676-8. ELShibini e¢ al “The Stability of Phenpleptvine—Put 2: The lscoluration reaction and the intvence of rome fons on the rate of sstopamine ant synephrine Br. Pharmacol, (1988), 93, tlradation of the dug” Arencimitelforehung. May 19655195) 828-9, ELShibiai, eal. “The Stability of Phenyephvie—Past 3: The racenistion reaction” Aeraeimitelforchung. Sep. 1909,199) 1613-4 ian eta, "The Stability of Aqueous Solutions of Phenylephine st Elevated Temperatures: Lent fenton ofthe Decomposition Pro tits” 1 Phanm, Phim, 1973.25. Sopp. 24P-31P, “Rept of the International Workshop on Vitro Methods for Assessing Acu Systemic Tis.” Ress ofa Ineratonal Work shop Orpaized by the latragency Coouinaing Commits onthe Valuation of Alteratve Methods (ICCVAM) an the National To cology Progam (NTP) Interagency Ceater for the Evaluation of ‘ternative Toxicological Methods (NICEATM), pp. 1370. ‘Zaczek, eal, “Te fect of phenylephrine on pain and fare inten. ‘ily in_oyee_ with iis” Acta Ophthalmol Scand, Ot 20078575105. * cited by examiner Primary Examiner — David J Blanchard Assistant Examiner — Barbs Fraziee (74) Attorney, Agent, or Firms — Wilson Sonsini: Goodrich S Rosati on ABSTRACT The invention is directed to methods and compositions of stabilizing phenylephrine fommations, The composition has cod time-dependent stability at low temperature and has 10 ‘change in its ourward appearance even after having been sored a least 6 months. 13 Claims, 4 Drawing SheetsUS 8,859,623 BI Sheet 1 of 4 2014 Oct. 14, US. Patent LaansiyUS 8,859,623 BI Sheet 2 of 4 Oct. 14, 2014 US. Patent ony oe eee ore ereinneepnennrnneaeeenioery 8 enszemr 9 peed ata Saalabec sinc ots sero “~ _& 'Zz0000e@0E).1 IvIbGNE!.L9¥) ov obaweu Bocenbts “a Lava v oppor PS 1€66 ‘ware vrs. Zaansiy @ % £66 = 22 ajdwes payspug AjpesuewenueugUS 8,859,623 BI Sheet 3 of 4 Oct. 14, 2014 US. Patent PE pS es Bo % €°66 = 08 aunyxq Brug eapoy eaundiyUS 8,859,623 BI Sheet 4 of 4 Oct. 14, 2014 US. Patent pound % 978 =88 ULIO}-S fqrpengooHenet LavwHa02! 1 9y) OO DOB4eY BOse=BIs 2 LavEUS 8,859,623 BI 1 METHODS AND COMPOSITIONS OF STABLE PHENYLEPHRINE FORMULATIONS BACKGROUND OF THE INVENTION Phenylephrine isa selective el-adrenergic receptor ago nist used primarily as a decongestant, as an szen toate the pupil, and to inerease blood pressure. Phenylephrine is mar- kote! asa substitute for the decongestant pseudoephedrine, though lineal studies differ regarding phenylephrine’s epinine>(+)-norepineph- Fine-(+)-epinephrine>(+)-phenylephine. ‘Two ophthalmic formulations, formulated 10% Phenyl phrine hydrochloride (S-isomer) an the exemplary invention ‘composition, 10% Phenylephrine hydrochloride (R-isomer)US 8,859,623 BI 1 were tested for their ocular activity in NZW rabbits. It was ‘observed that formulated isomer showed minimal dilation, responded to light exposure and constricted sliyhily more slomiy thn the untreated eye, where as the exemplary inven- tion composition, 10% Phenylephrine hydrochloride showed ‘maximal dation with in 15 min of dosing and the pupil di not espond 1 ight and remained dilated for 4 hrs. “According to the above study it could be postlated tha, ‘when an ophthalmic solution of phenylephrine hydrochlo- ride, (R-isomer) containing S-isomer as an impurity is used Jor dilation of pupil, the s-isomer may cause te saturation of the wadrenengic revepiors resulting in the decrease in the response ofthe cg after its administration (taehyphylsis). Furthermore, the presence of S-isomer in the ophthalmic solution may lead to pooridelayed dilation of the popil In some embodiments provide methods of dilating the pupil comprising administering a composition comprising R-phenvlephrine hydrochloride topically to a mammal ‘wherein the composition substantially maintains the initial chiral purty of Rephenylepbrine hydrochloride fora least 6 ‘months. I is evident from the literature that the pharmaco- Jogieal evaltation of both R & S-Phealepirine hydrochlo- Fide is not same. R-Phenylephrine is referenced as wscfl synthetic adrenergic drug Uveitis Uveitis is, broadly, inflammation of the uvea. The wvea ‘consists of the mide, pigmented, vascular structures ofthe ‘exe and includes theirs, ciliary body, and choroid. Uveitis reajires an urgent referral and thorough examination by an ‘ophthalmologist or Optometrist and urgent neatment to con= trol the inflammation, Anterior wits (iii) afects the front portion of the eve, intermediate uveitis (eyes) affects the Ciliary body. and posterior uveitis (choos) affects the back portion ofthe wea, Diffuse uveitis alfeets all portions of the uvea. Anterior uveitis commonly occurs in conjunction with juvenile rheumatoid aris, but does not manifest inall juvenile anhrts patients. Uveitis is most ikely #o be present ‘njuvenile arthritis patients with pauciaricuae disease (Feiee than five joints involved), a positive anti-nuclear antibody test, and a negative rheumatoid factor test Tt has been dem- ‘onstrated that ater phenylephrine hydrochloride ophthalmic solution instillation, flare intensity and pain were signifi ‘cantly decreased only in eyes with irdocyelitis and without ‘Abrinoid maetion (FR). The decreasing level of are intensity, ‘and paralysis of the pupil aller phenylephrine instillation seem (alleviate pain in those eyes. See eg. Zaezek, et.al. Acta Ophihalmol Scand. 2000 October, 73):516-8. In some embodiments provide methods of eating Uveitis, jn a subject comprising administering a composition com- prising R-phenylephrine hydrochloride 10 sad subject, ‘wherein the composition substantially maintains the initial chiral purity of Rcphenylephrine hydrochloride fort least 6 months, Tn some embodiments provide methods of performing eer- ‘win ocular testing such as ultsonograpiy, provocative ‘closed angle glaucoma test, Retinoscopy, compromised cir- culation (.., blanching est), Refraction, fundus examination ‘comprising administering a composition comprising R-phe- rylephrine hydrochloride, wherein the composition subst tially maintains te inital chiral purty of R-phenylephrine hydrochloride for at last 6 months Tn some embodiments provide methods of aiding surgical procedires requiring visualization ofthe posterior chamber ‘comprising administering a eommposition comprising R-phe- raylephrine hydrochloride, wherein the composition substan- tally maintains the inital chiral purty of R-phenylephrine hydrochloride for at least 6 months 0 o 8 After preseatation of R-phenylephirine hydrochloride oph- thalimie solution 2.5% or 10% to the ocular surface, a broad variation inthe delay of onset of dilation is widely reported, varying between 20-10-30 minutes and as mach as up to 60 ‘mines, Whilea number af coneiburorsto this delay of onset have been theorized, the absence of phenylephrine kydro- chloride's pharmacologic activity inthe eye dve to the pees- fence of S-pheaylephrine may in fact be the explanation for such delay Dropper Bottle or Storage Bottle Conventional dropper botles for administering oph- ‘halle Hud are well awa inthe prior art, The basic com- mercial design of such dropper bottles has remained fairly ‘unchanged over the last several decades: a squeezable eo tainer is provided with tapered dispenser that terminates in discharge aperture, To administer ophihalasie Hid, the di charge aperture is aligned above a taruet eye andthe bot is squcezed tounge outa drop or dose ofthe Hai, Alternatively, liquid dispensers have been developed in whieh the formulation is supplied from a stomge botle through a dropper, for example (dropper bottles or FIDO- COphiiols), The aqueous formation usually ows out of the tdmpper opening a a result of manual pressire being applic to the compressible storage botle. Insome embodiments, the composition described herein is stored ina plastic or glass botle. In certain embodiments, the plastic bottle is a low-density polyethylene bottle. In certain ‘embodiments, the composition deseribed herein is stored ina lass bottle with or without a liquid dispenser. In certain ‘embodiments, the plastic or glass hotle is opaque, ‘Additionally, the compositions described herein are ether packaged for single use o for multiple uses with or without a preservative Certain Pharmaceutical aad Medial Terminology The term “acceptable” with respect toa formulation, com- position or ingredient, as used herein, means having no per- sistent detrimental eect on the general health ofthe subject being treated. ‘The tem “cari as sed herein refers to relatively aon ‘oxie chemical compounds or agents that fseltate the inéor portion af «compound into cells or tssues. The terms “co-adminstration” of the like, as used herein, are meant to encompass administration ofthe selected then peutic agents to a single patent, and are intended to inelude {reuiment regimens in which the agents are administered by the same o different route of administration orat the same oF illo time. ‘The tem “diluent refers to chemical componnds that are used to dilute the compound of interest prior to delivery. Dilients can also be used to stabilize compounds because they can provide a more stable environment. Salts dissolved in bufered solutions (which also ean provide pH control or ‘mainlenance) are utilized as diluents in the ar, including, but ‘ot limited to a phosphate bfered saline soltion "The tems” therapeutically effective refer to a sufficient amount of an agentoracompound being administered Which will rieveto some extent one oF more ofthe symptoms of the disease or condition being tested. The result can be reduetion andor alleviation of the signs, symptoms, or causes ofa disease, or ‘any other desired altertion of biological system. For texample, an “effective amount” for therapeutic uses is the Amount of the composition comprising 2 compound as di closed herein required to provide a clinically signifieant decrease in disease symploms. An appropriate “effective” ‘amount in ny indica case may he determined using toch- ques such as a dose escalation studyUS 8,859,623 BI 9 The terms “enhance” or “enhancing,” as used here means o ineease or prolong ether in potency or duration & ‘desired effect. Thus, in regard to enhancing the effect of therapeutic agents, the term “enhancing” eles tothe ability twinerease or prolong, cher in potency or duration, the ellot ‘of other therapeutic agents on a system. An “enhancing ‘effective amount,” 36 used herein, refers to an amount adequate to enhanee the effect of another therapeutic agent in a desired system, The term “subject” or “patient” encompasses mammals Examples of mammals include, but are not limited (0, any member of the Mammalian class: humans, non-human pri- mates scl as chimpanzees, and other apes and monkey spe- ‘ies: farm animals uch as cattle, horses sheep, goats, swine: ‘domestic animals such as rabbits, dows, and eas; laboratory animalsincluding rodents, suchas ats, nice and guinea pigs ‘andthe like. In one embodiment, the mana is « human. aileastonesymnp= tom of a disease or condition, preventing additonal symp- toms, inhibiting the disease or condition, ea, agresting the devclopment ofthe disease or condition, eicving the disease ‘or condition, causing regression of the disease or condition, relicving a canition caused by the disease or condition, of sopping the symptoms of the disease or condition either prophylactically and/or therapeutically ‘Allof the various embodiments or options described herein ‘can be combined in any and all variations. The following Examples seeve only to illsteat the invention and ae not 10 be constried in anyay to Fimit the invention, EXAMPLES Example 1 Exemplary Phenylephrine HCI Ophthal Formulation R-Pheayleprine Hydrochloride Ophthalmic Solution, USP 2.5% oF 10% isa sel, clear, colorless to light yellow, ‘opical mydriatie agent for ophthalmic use. The chemical name is (R)-3-hydroxy-<{(methylamino)methylJbenzen- ‘emethanol hydrochloride. R-Phenylephrine hydrochloride is represented by the following structural formula \ sc ts Phenylephrine hydrochloride has a molecular weight of 208.67 and an empirical formula of C,H, .NO, HCL Fach mL of R-Phenylephrine Hydrochioride Ophthalmic Solution, 2.5% contains: ACTIVE: phenylephrine hydro- chloride 25 mg (2.5%); INACTIVES: sodium phosphate ‘monobasic, sodium phosphate dibase; bore acid, water for injection. Hydrochloric acid and/or sodium hydroxide may beaded ro adjust pH (6.0 10 6.4). The solution has a tonicity ‘0500 mOsm/kg; PRESERVATIVE: henzalkonium chloride 0.01%, Fach ml. of R-Phenylephrine Hydrochloride Ophthalmic Solution, 10% contains: ACTIVE: Rephenylepbrine hydro- chloride’ 100 mg (10%): INACTIVES: sodium phosphate monobasic, sodium phosphate dibasic; water for injection. 0 o 10 Hydrochloric acid andor sodium hydroxide may be aed to adjust pH (63 (0 6.7). The solution has a tonicity of 1000 ‘mOsm/kg: PRESERVATIVE: benvalkonium chloride 0.01%, ‘Tae composition of Phesylephrine HC] Ophthalimie Sokt- tion, 2.8¥% and 10% is iste in Table 1 ‘Table 1: Phenylephrine HICI Ophthalmic Solution, 2.5% and 10% Quantitative Composition 210 Qusty “Qussy ust oan Been) ows) Funston Suaatd Soham 08 6k Std igo Asmat Amend pause Example 2 Stability (Impurity) Test and Results Stability studies of 2.5% and 10% Phenylephrine FICL solutions prepared as in Example 2 were conducted at 210 8° for 12 months ‘While the resting performed during the historical stability ‘analysis limited, those parameters evaluated show excellent results, For the 3 batches of 25% formulation evalusted, the inka assay averaged 101.2% of label claim (range 99.8%. 102.9%).and after 12 months of storageat the labeled storage condition (2-8 C.) the average potency was 99.7% of label claim (range 97.0%%-103.4%). All other parameters evaluated (ppearance, preservative effectiveness, sterility) conformed to specifications. For the 3 batches of 10% formulation evaluated, the initia assay averaged 100.4% of label claim (range 99.8% 101.626), and afer 12 months of storageat the labeled storage condition (28° C.) the average potency was 99.8% of label claim (range 98.8%-101 0%). All other parameters evaluated (eppearance, preservative effectiveness, sterility) conformed to specifications. Example 3 (Chieal HPLC Analysis ‘The following are non-limited exemplary chiral columns and relevant mobile phases in the methods for analyzing chiral pority of R-phenylephrine Coftimn-O1-RH (150x6) mm, $m, Flow: Im min ‘Mobile Phase: Methanol, Column Temp: 25° C., Detection ‘wavelength: 270 nm. Column-OJ-RH (1503.6) mm, 5m, Plow: 0.8 mL. min ‘Mobile Phase 0.05% Fthylenediamine in Methanol, Columa “Temp: 25° C, Detection wavelength: 270 nz. Cofumn-O1-RH (1504.6) mm, Flow: | ml min-1, Mobile Phase: 0.05% Ethylenediamine in Methanol, Colma Temp: bient, Detection wavelength: 270,US 8,859,623 BI u Column-OJ-RH (150x446) mm, 5 wm, Flow: Lm min, Mobile Phase 0.05% Ethylenediamine in Methanol, Cluma Temp: 25° C., Detection wavelength: 270 nm. CColumn-O1-RH (150x446) mam, 5 um, Flow: 1 ml min Mobile Phase: 0.05 Ethylenediamine in Meduanol, Column “Temp: 25° C., Detection wavelength: 270 am, Cofumn-OJ-RH (150%4,6) mm, 5 im, Flow: 1m min, Mobile Phase: 0.05% Flhylenediamine in Water (05%y Methano! (95), Column Temp: 25°C, Detection wavelength: 270 Column-OJ-RH (150x446) mm, 5 wm, Flow: 1m min, Mobile Phase 0.05% Ethylenediamine in Methanol, Columa “Temp: 25° C., Detection wavelength 270 am, Column-O1-RH (150x4.6) mm. 5 um, Flow: Lm min “Morile Phase 0.05% Ethylenediamine in Methanol, Columa Temp: 25° C., Detection wavelength: 270 am. Cofumn-OJ-RH (1504.6) mim, 5 un, Flow: 0.5 mi min, Mobile Phase: Acetonitrile: 0.05% Fihyleneiamine in water (30:70) Columa Temp. 25° C., Detection wavelength: 270 Column-OJ-RH (150%4.6) mm, Sym, Flow: 0 ml mil Mobile Phase: Acetonitrile: 005% Ethylenesdiamine in water (40:60) Column Temp.: 25, Detection wavelength: 270 am, Cohumn-Chiralpak IC-3(150%4.6) mm, 3 um, Plow: 1. Oma min-1, Mobile Phase: 0.1% Fthylenediamine in n-Hexane (85%): Ethanol (15%), Column Temp: 25° C., Detection ‘wavelength: 270 um; re 600 nm, Cohumn-Chiralpak C-3 (1504.6) mm, 3 ym, Flow: 12m min-1, Mobile Phase: 0.1% Ethiylenediamine in n-Hexane (50%): IPA (50%), Column Temp: 25° C., Detection wave- Jength: 270 nm, Columm-O1-RH (150x4.6) mm, Flow: 06 ml mil Mobile Phase 0.05% Ftkylenediamine in Methanol, Column “Temp: 25° C; Detection wavelength: 270 nm. 4.0mg sample in mL-ethanolwas analyzed. The injection volume to HPLC §s 3.0 ul. The HPLC chromatogram is shown in FIG. 1 The HPLC chromatogram clearly show separation of race mic sample. Chiral HPLC method was this established 10 ‘analyze Phenylephrine Pxample 4 Determination of Chiral Purity ater 6 Months Storage at Low Temperature R-Phenylephrine Hydrochloride Opthalmie Solution, 2.S¥oand 10% prepared as in Example | Wer stored at 210 8° ©. The chiral purity of Sample I (10% solution) was assessed before low temperature stability test, The HPLC chromato- ram is shown in FIG. 2. “The chiral purty of R-Phenylephrine Hydrochloride was determined by the method and conditions as showin in Example 3, The result showed 99.3% ce ‘After 6 months of low tomperaturestorage (ie. 2008°C:}, the chiral purty of R-Phenylephrine Hydrochloride in the solution was detemined to be 99.3% ee. The HPLC ehro- Toconfinn the “impurity” shown in the chromatogram, the “impurity” was purified and determined bythe same method. ‘The “impurity” (i.e, S-Phenylephrine Hydrochloride) was determined to possess 82.4% ee oF Storm. The HPLC chro- matogram is shown in FIG. 4 ‘Thus, itis leary shown that the Solution remain substan- tially maintains the inital chiral purty of R-phenylephrine hydrochloride for at least 6 months 0 o 12 Example 5 Dilation Assay of S Form Phenylephrine Solution Both R and S form solutions (10% solution prepared asin Example 1) were test for dilation on rabbits. The first test rabbit received 3 drops of the S form formulation and the second test rabbit received 3 drops ofthe R form solution. “The resulls wore as follows Test Rabbit No, 1: Minimal Dilation, within 15 minutes of| ation the pupil was only slightly more dilated than the untreated eye. The treated eye responded to light exposure and constricted slowly. The control eye constricted rapidly as Was expected “Test Rabbit No 2: Maximal dilation within 15 minutes of dosing. The pupil did not respond to light exposure and romsined fully dilated for 4 hours thea regressed. ‘These results clearly show that an ophthalmic solution of phenylephrine containing S-isomer does not dilate the rabbit pupil as it is achieved with an ophthalmic solution of phe- rylephrine containing R isomer. Ths itis evident that main- ‘aining te chiral purity ofthe ophthalmic solution i erucal to keep drug potency. ‘While preferred embodiments of the present invention hhave boon shown and described herein, it will be obvious 10 those skilled in the aft tht such embodiments are provided by ‘way of example only. Numerous variations, changes, and substinutions will nw occur to those skilled in the art without Sparing from the invention, It should be understood that various alternatives 10 the embodiments of the invention scribed herein may be employed in practicing the iaven- ‘ion, Its intended thatthe following claims define the seope of the invention and that methods aad stractures within the scope of these claims and their equivalents be covered thereby. ‘What is claimed is 1. A method of wsing an ophthalmic composition for pupil ilation, the composition comprising R-pheaylephine faydrochloride having an initial chiral purty ofa least 95% and an aqueous butter, wheroin the chiral purty of R-phe- sylephrine hydrochloride i at least 95% ofthe inital chiral purity afer 6 months, the method comprising: ‘administering the composition into an eye of an individual in need thereof, wherein the composition is stored between =10 to 10 degree Celsius prior to administra tion, and wherein the composition comprises R-phe- nylephrine hydrochloride having a chiral purity of at least 95% when administered afer storage. 2. The method of claim 1, wherein the composition is allowed to be stored between 2 10 8 degree Celsius 3. The method of claim 1, wherein the composition com- prises R-phenylephrine hydrochloride having an intial chiral purity of atleast 99% 4. The method of claim 1, wherein the composition com- prises R-phenylephrine hydrochloride having an intial chiral purity of at least 99.3%. '3, The method of claim 1, wherein the chiral purity of phenylephrine hydroclorde is at least 979% ofthe initial chic purity alter 6 months. 6. The method of elaim 1, wherein the chiral purity of phenylephrine hydrochloride i at least 992% ofthe initial chiral parity after 6 months "7. The method of claim 1, wherein the chiral purity of phenylephrine hydrochloride is atleast 99.5% ofthe int chiral purity after 6 monthsUS 8,859,623 BI 13 8, The method of claim 1, wherein the composition co prises 2.5% wiv of 10% w/v Rephenylephrine hydrochloride by weight '9. The method of claim 1, wherein the composition is packaged in a 1-15 mi plastic or glass bottle 10. The method of claim 9, wherein the pockage identifies storing the compositionata temperature between = 10,0 10C. 11. Themethod of claim 10, wherein the package identifies storing the composition at a temperature between 2 108 C 12, The method of claim 9, wherein the composition isin & plastic or glass bottle of about 2m, about 3m, about Sm, ‘about 10 ml or about 15 ma 13, The method of claim 9, wherein the plastic or glass boat is opaque 14