AU2013202641B2 - Stable sns-595 compositions and methods of preparation - Google Patents

Abstract

Methods of preparing substantially pure SNS-595 substance are disclosed. Also provided are compositions comprising SNS-595 substance that are substantially pure and essentially free of visible particles. Eg o4 -i E 0.00% 0.05% 0.10% 0.15% 0.20% 0.25% Compound 4 (% w/w) in Voreloxin Drug Substance

Description

STABLE SNS-595 COMPOSITIONS AND METHODS OF PREPARATION [0001] The entire disclosure in the complete specification of our Australian Patent Application No. 2010289337 is by this cross-reference incorporated into the present specification. 10002J Methods are provided for preparing substantially pure 4 +dihydro [(3,48)3mehoxy~4~(meyamino)-1 yrro lidinyi.4~oxo- (24hiazoliy)~8 naphthyridincearboxylic acid, together with eomposi tion s conmprising tis substance. 2. BACKGROUND) j0003) Thbe compound ( 1 i4dihydroz74(3S4S)3methxyw-mthylamino- I pyrroidinyl4oxo- I (2thiazolyl)- ne o acid, has the faIowing cheuncal structure; COO H- CHN* N N N S, N ACLOC This compound is also known as SNS-595 or AG- 352 Tbhe United States Adopted Names Conl (USANC) has assied the name "vosaroxin to ,Is con poud, [00041 SNS-595 is known for it ant-tumoracti wee Tsuzuk ci et aJ ed C he 47,2209 06, 2004 and Tomita et al MJ. Che n4555645575, 2002) Treatment of vaniousa cancers with SNS-95 has been proposed in the literature and SNS-59 5 has shown prechical actxity agamatIvarious cancer ceJifes and x nografls Various dosuig regimens for the use of this compound have been reported. For example.see US Patent Application Pub. Nos 20050203 120 A1 2005-21 S3 AL and 2006 0025437 A I, all of which are incorporated herein by reference in their entireties: SNS 595 is presently being tested in inica trials to assess safety and efficacy in human cancer patients and has demonstrated cliniaactivity in the treatment of acute myveloid tekemia and ovarian. cancer. 41- [0005] SNS-595 can be prepared using techniiques known tO those of ordinary skill in the art Se. tor xampileRS5 Patent No, 5,817,669. issued October 6, 1998 lpanese Patent Appldation No IHei 10-173986, published une 26, 1998; WO 200746335, published December 21, 2007; Tsuzuki et al J Me. (Them, 47:2097210t 2004; and Tomita etal j MedCan,45:5564557552002, alo whiCi are incorporated herein by reference in their entireties: [00061 Intemational Patent A pplication No. WO 2007/146335 describes an exemplary process tor the synthesis of SNS 595. As shown in Scheme 1, this synthesis proceeds via intermediate , which is reacted with Compound 2 in the presence of a base,sucha triethylamine or N N-iisopropylethylane to form Compound 3, The ester group of Compound 3 is subsequently hydrolyzed to afford SNS-r595 3N~s The route de inScheme I hor yi ds idesrabeO of impuitis ien y-roducs of the reactons. hich arecif, dtt deplete orremoe from the SNS~595 drgsubstance and SNS~595 drgprodu [00071 Athuheranb rdutcaeisinN-95peatinreu gte amount of the-se impurit ies in. the-, drug substance c and the final dt-gl product is"- hygport-art. Since cancer patients u-ndergo significant hentrayand radiation therapy and er thereforehvecmroie imne systems, it is beneficia[ o ddliver to these aiet drug product that is characterized by high ptyFurther-, for intravenous orinareia admnisratonof a druIg prodet, the pudty and physL datrsics of the drug product are important because the drug product ente-rs dietyinto th blodst!,re-.am 100081 Thus, there remains a need for improved methods for pre.parin SNS -5 subtaniaHy fRee of Contaninants, therebly providing adrgsbaneiasutntly Pure fomt decib ed i Scfo 1.hor formulation into pharmaceutical p cts fort r of caner wihou ths need for laborious. o puarifiiiopnnd threorhae omrmiedimue ysemitisbeeicalo eivr o hsepaiet I. SUMMARY [00091 Processes are provided that can ield a substantial pure SNS-595 substance, In addition, the processes can be scaled upzto commercianufacturin of substantially pure SNS9-5 substance. 10010] In some enilbodiments, processes are Provided for preparing a substantial pure SNS-595 substance comprising: (a) reacting Cornpound 3 02E MVeHN-/-N N S MeNO V4 3 with a first aqueous base folowed by utaliig to obtain a primary SNS~ 595 hydrate: (b) dehydrating the primaryN S595 hydrate fromri step (a) and reacting the dehydrated product with a second aqueous base followed by neutralizing, to obtain a secondary SNS-595 hydrate; and (e) dehydrating the secondary SNS595 hydrate obtained in step (a to obtain the substantay nure SNS595 st stacoe [00111 In some cases it may be desirable to react the substantially pure SNS 595 substance obtained in step ) with a further aqueous bas5e neutralizes tn dtehydrate to furthey improve the purity of the substanially pur SNS-595 product. The first aqueous base, the second aquous base, and the further aqueous base(s) can be the same or diterent Likewise the acds employed during the neutralinion steps may be the same or different. Recycltng of the substantially pure SNS-595 substance to gh the described steps of treatnen vith aqueous base, neurzation mand dhydratonay be performed a pluraity of times to sequentially further purify the SNS-595 substance until a substantially pure SNS-595 substance of a doesred levo purf obtained. f0021 In. some embodiments processes ar e provided forpreparig a substantial ly pure SN S15 substance, co. uprising: (a) reacting Compound 1 C.. t .N N with Conpound 2 Me, NHMe 2TsOH N H 2 in the presence of N-diisopropyletai and water to obtain substantially pure Conpouud 3 XCOEt N N N MeHN.(Y r 'N N S Med G 3, and (b) reacting Compound 3 vib an atueous base to fbrr te substantiug pure SNS-59$ substane, [00131 In som. embodirnets. provided herein are comlposions comprising substanially pure SNS-S95 substanc, wherein the substantially pure SNS-595 substance compnscs about 0 to 001% Compound 4 OHO S N 4 at the tine of production of the compositions, based on total weight of t sustantian pure SNSW95 substance.

[00141 In some embodiments, provided herein are compositions comprising substantially pure SNS-595 substance, wherein the substantially pure SN ~595 substance comprises about 0 to 0.0.2% Compound 5 0 co CCA }i T 4'OH N N at the time of production of the compositions based on totalweight of the substantially pure SNS-595 substane.: (00151 In some erbodinents. compositions are provided that comprise SNS-595 and water, wherein bout 100 mng of SNS-595 is present for every 10 mL of fihe compositions, wherein the compositions are essentially f-ee of visible particles, and wherein the compositions arc stable at for example, 3, 6, 9. 12, or 24 months after production 100161 In some embodiments, compositions are provided that comprise SNS~$95 and water, wherein about 100 mg of SNS-595 is present for every I10 mL of the compositions, wherein the compositions are essentially free of sub-visible particles, and wherein. the compositions arc stable at, for example 3, 6, 9, 12, or 24 months after production, [001,71 In some embodiments the processes described herein are performed on a kilogram seale. 4. BRIEF DE SCRIPTION OF ThE DRAWINGS [0018.1 Figure 1 illustrates the observed relationship between amounts of Compound 6 in SNS-595 bulk drg product solutions formulated from drug substances having different amounts of Compound 4, 100191 Figure 2 illustrates the observed relationship between the amount of Compound 4 in SNS~595 drug substance and the amoun of residual Compound in Iompound 3 used to prepare the drug substance. t00201 Figure 3 illustrates the observed relationship between the amount of Compound 5 in SN-595 drug substance and the amount of residual Compound I in Compound 3 used to prepare the drg substance -5- 5. DEAM t.El !LD )ESCIU PTION 4.1 DEFINITIONS [00211 Unless defined oderwis, all technical and scientific 'run used herein have the same meaning as is commonly understood by one of ordinary skill in, the art, All patents, applicatins published applcauons and other publications menltCed herein are nleorporated by r nene in their entirety. 100221 As used herein, SNS-595 reers to (i)I4-dihm metnhy laminopyrrohdminy]4~oxo S( thioly 18 naIphthyrdne& Caboxylic acid, as well as any ionic forn. sals so\vates og, hydrates or other forms of that compound, inchiding mixtures thereof, Thms ornpositon~s comipising SNS-595 iincIide (t) 44 dihydro-4(3$,47)-ethoxy &methyhuanin.o1 .pyrrolidinyflp4eoo4(-thuazolyl) 1;8 n -hhvii -~carbosv ie acid or, in sole erubodhneents, an ionic form throf sa solvate, e.g hydrate. polymorph pseudomrph tor other foor ofthe compound, ]n some enbodiments. SNS495 is provided as a pharmaceutical acceptable salt SN S95 is also referred to as AG-7352. voreloxin. and. vosaroxin. 100231 As used herein. "substantially pure SNS-595 substance" refers to a composition conisting essentially of SNS-595, .. \comprisfig less than about 1 % ofa an other individual cionmpound (or impurity based on total weight of the compositionfAt%) For examiple in some embodiments, such compostions comprise about 0 to 0 5% about 0 to 0, 1% about 0 to W05%. about 0 to 0 03% about 0 to 0.02%, or about 0 to 0 0 Compound. 4 based on total weight of the composition. OH 0 i i Cr IN 4 In. some ernbodirnents such compositions eonstessentially of 0 to 03%, 0 to 0. 1. 0 to 0,05% 0 to 0O3% 0 to or02%KOr 0 to 0.0 Compound 4 based on total weight of the K''0 to 0 %9to Q..'Xt 0 to comiposition. Insom enmbodinmentis.such copstin ave .. to 0 5%0 o 1 t 005%, 0 to 0,03%, 0 to X0,2% or 0 to 00 Compound 4 based on total weight of the 6~ composition. In some embodinments. the comlpOsiOns have _0 02% of Compoun d 4 based on total weight of the Comnposi tion In some embodients suchcompositions comprse abota 0 to 0.04%, about 0 to 0,03%. or about 0 to 0,02% Compound 5 based on total weight of the composition. 0 0 CK N 'N N \=/ In some embodiments, such compositions consist essentially of 0 to 0:04% 0 to 0.03% or 0 to 0.02% Compou-nd 5 based on total weight of the composition. In some embodiments, such compositions have of 0 to 0,04%, 0 to 0 3% or 0 to 0.2% Compound 5 based on total weight of the composition, i some embodiments, the compositions have $0.15% of Compound 5 based on total weight of the conosition Other compositions that arc substantially pure SNS-595 substances are described herein In some emibodimentsuch compositions comprise about 0 to 0 out abou - 0 to 01%, about 0 to 005%, about 0 to L03% about 0 to 0.02% or about 0 to 0,01% Compound 6 based on total weight of the composition, OH OH Ci N 1 0N N- Cl SC N N N 8 VJ 6. in some embodiments, such. coropositions consist essentially of about 0 to o.%, about 0 to 0. 1%,abt out 0 to 0.05%. about 0 to 0 ao r ow0.01% Compound 6 based on totai weight of the composition, in some embodinents such compositons have about 0 to 03 % about 0 to 0. 1%, about 0 to 0.05%. about 0 to 0.03%. about 0 to 0.02%, or about 0 to 0,01% Compound 6 based on total weight of the composition.

in some embodients, such compositions comprise about 0 to 0.5% about 0 to 0. 1% about O to .05%, about 0 to 0 03%, about 0 to 002%or about 0 to 0.01% Compound 7 based on total weight of the composition HQ N N -J 7. In some embodimients, such compositions consist essentially of about 0 to 0.5% about 0 to 0.1%, about 0 to (105%, about 0 to 0133, about 0 to 0.02% or about 0 t 0,01% Compound 7 based on total weight of the composition. In some embodiments, such conposinons have about 0 to 0:5%, about 0 to 0.1% about 0 to 0.05% about 0 to 003% about 0 to 0.02%, or about 0 to 0,01% Compound 7 based on total weight of the composition. In some embodiments, the compositions have CO0 1 5% of Compound 7 based on total weight of the composition [00241 As used herein. SNS95 substance" refers to a composian comprising SNS 595 and one or more other compounds .In some enbodiments the S:NS-595 substance comprises SNS-595 and Compound 4 andor Comrpound 5 j00251 As used herein and unless otherwise indicated "about" refers to up to plus or minus 10% of the indicated value. For example, aboutt 0.01 %" refers to 0.009% to 001.1% "about25 IC" rebrs to 225 'C to 27Y "C, and "about 0 6 M" refers to 0.54 M to 0.66 MY. In some embodiments. about refers to up to plus or minus 9, h. 7, 6, 5, 4, 3. 2. or 1% of the indicated value. Similarly, for a range of values, use of "about" refers to both the upper limit and th cdower limit of the stated range. [00261 The tem"about" wih respect to a pH value is intended to mean that the acceptable error fotr that pH1 value is no greaterthan 0.1, 0.02. 0(13 0.04. 005 0,0 007 0.08, 0.9, or 01 pH unit. In certain embodiments, the error for a pH value is no greater than 0.02 pH unit (see, Method 791 in USP XXVI (2003) incorporated herein by reference in its entretvy 100271 As used herein "aqueous base' refers to any aqueous sorion o One or more bases, which, in some embodiments, are one or more strong bases (pKa ' 13) Examples of -8strong bases include.without limitation. hydroxides of alkali and alkaine earth metals or ammonium. hydroxide Aqueous bases may be aqueous solutions of organic or imorganic bases. i some embodiments the base is provided as an aqueous solution of poTassium hydrox ided ithium hydroxide, sodium hydroxide, or ammonium hydroxide, In some embodiments. the aqueous base has a. molar ooncentrationot about O-1. . M. In some embodimnts, the aqueous base is an aqueous solution of a hydroxide hwing a molar concentration of about 0,6, 0,7, 0,85 0,9, L 0 or about 1,1 Mi In some embodiments, the base may be provided in solid fon. in. some embodiments, the solid form is a pellet or a powder, [00281] As used herein and unless otherwise indicated, the term "hydrate" means a compound or salt thereof. further including a stoichiometric or non stoichiometric amount of water bound by non-covaien t intermolecularforees. As used herein and unless otherwise indicated, the term "solvate" means a solvate formed from the association of oer more solvent rnoleeus to a compound provided herein The term "soIvate" includes hydrates (e~g., monohydrate, dihydrate, trihydrate, tetrahydrate, and the ike). The solvates of SNS 595 can be crystalline or non-crystaline. 100291 As used herein. "SNS595 hydrate" refers to SNS-595 having astoiimetri or non-stoichiometric amount of water bound by non-covalent intermolecular tore in some embodiments, the SNS-595 hydrate is crystalline or non-ervstaliine. In some embodiments, the SNS595 hydrate comprises about 00to 12 molar equivalentsof water per mole of SNS-595. in some embodiments, the SNS-595 bdrae comprises about I molar equivalent of water per mole of SNS-595. [00301 As used herein "dehydrating" refers to removing a:ir bound to the SN$-595 of a SNS-595 hydrate, Dehydrating metbods are known to those of ordinary skill in the art, In somenbodients, dehydraung is accomplished by contactin the SNS9 hydra with a compound capable of removing water bound to the SNS-595 of the SNS-595 hydrate, Such compounds include dehydrion solvents, In sone embodimentsthe solvent ishygroscopic and/or proie. Exemplary solvents inclhide, without limitation, methanol. ethanol, isopropanol, acetone or others apparent to those of ordinary skill in the art. In some embodiments, the solvent i~s anhydro us. In some embodiments, th~e solvent is anhydrous ethanol. In some embodiments, e anhydrous ethanol comprises less than 0.5% water; in a -9~ particular embodiment, dehydrating is accomplished by contacting SNS-595 hydrate with anhydrous ethanol at about 25-0 "C, about 40-80 'Q about 6~80 *C' or about 80 WC. I some cases, dehydration may be accomplished by thermal means in the absence of a solvent, [0031] The amount of water in a hydrate may be anal.ed using a number of techi piues as understood in the art For example the amount ot water may be determined based on the observed weight loss atheroravietric analysis G rogran In e exhaust from a TGA furnace na be coupled to an instrument of chemical analysis.such as a mass spec trometry instruIent or an infrared spectroscopy instrument, to conmfir the chemical purity of the water vapor emited upon heating, Water loss may also be quantified by direct gravimetric means such as "Loss on Dying <73I" as described in the US, Pharmacopoeia, th e entirety of which is incorporated herein by reference, Karl Fischer KF) analysis may be used to analyze the water content of a hydrate smeCoulometric KF analysis for water determinatJon may be performed using a Mettler Toledo DL39 Karl Fisher titrator or otner equipment. In one method; approximately 1402 mg of a sample is placed in a K tiration vessel contang H-YDRANA,, @S ouloa t AD reagent fbr coulometrie RE titration and mixed for 60 seconds to ensure dissolution The sample is then titrated by means of agenerator electrode. which produces iodine by electrochemical oxidkationThe analyss is repeated to reproducibility of th [00321 As used herein -neutralizinf or "neutralization refers to the process of adjusting the pH of a sohtion to neutrality or approximate neutral ity; e-. p-I of from 6.0 to 8,0, 7.0 to 8.0, or 73 to 71. 100331 As used herein, "particulate matter refers to any matter formed as a resu of by products of a synthetic preparation of SNS-595 that is insohible or partially soluble in water or an aqueous mixture In some embodiments, the particulate matter comprises visible particles. Other impurities including articulates such as lint, glass, metal, and the like may be present in the SNS-595 compositions at or below levels permitted for administration to human subjects in the treatment of disease. [00341 As used herein visible particles" refers to insoluble or partially soluble solids in a iquid sounton. e.g. an aqueous solution, that are visible to a h umnan ee, In some embodhments, the visible particles are visible under natural sunlight white-light, fluorescent 10ightin. or incandescent lighting. in further embodioents, the visible particles are visible when observed under lighting h a n s of 600-000 lux, 900-4000 lu, 850-4650 lx, or 2000-3750 Ix n some embodiments, the particles are visiie when inspected for about160, 1-30, 15 1-10, i5, or 5 seconds, [03] In a patclrebdmn;te visible pardicles are visible when tested according to the method described by European Pharmacopeia 5.0, Section 192.20, the entirety of which is incorporateid hrein by reference In this method, an apparatus having a viewing station comprising () a matt black panel of appropriate size held in a verical position; (2) a non-ghre whine p-ane ot appropriac size held in a vertical positionext to the black panel; and (3) an adjustable ampholder fitted with a suitable. shaded, w hite-light source and with a suitable light diffuser (e ga viewing illuminator containing two 13 W fluorescent tubes, each 525 mm in length) is used The intensity of illumination at the viewing point is maintained between 2000 lu and 3750 la Higher veues may be used for glass and plastic contaies Adherentabels are removed from the containercontaining the sample to be tested The outside of the container is washed and dried, The container is genly swirled or inverted while ensuring that air bbles are not introduce, adc the container is observed for about 5 seconds in front of the white panel to determine whether visible particles ae presn The container iS then observed for about seconds in front of the black pane to de termine whether visible particles are present. In some embodiments, the visible paticles are visible aaimst a white panel In some embodiments, the visible particles are visible against a black panel. [00361 In some embodiments the visible particles have an average diameter of at least 50 gim, at least 75 am at least 100 pm at least 150 pm., or at least 200 pm In some embodiments the visible particles have an average diameter of about 50-500 am. about 50 300 pn, about 100-500 pn, or about 100-300 urn 100371 As used herein. "sub-visible particles" refers to particulate matter detectable by theight Obscuration Particle Count Test or Microscopic Particle Count Test described in the U S. Pharmacopoeia, -78> Particulate Matter in Injections, which is incorporated herein bv reference in its entirety 100381 In some embodiments, the visibIe and/or sub-visible particles cornprise matter frm the production process of a drug substance composition, including, without imitation, -1lP the synthesis or formulation of the drug substance, or the packaging of the composition In some embodiments, the visibLe parties compose metaL glass, lint or the like 100391 In certain embodmen s. the visie andor subvisible particks comprise one or more of Compound 5 o a NL IKF OH C I N N N \=/ 5, and/or Compound. 6 OH OH S-N N S ' C ' N' NI 0 m NI N CI NS 6 and/or Comnound 7 0n0 7 [0040] As used herein essentially free of visible particles" refers to a iquid, e'g., aqueous, composition comprising SNS-595 that does not contain visible panicles In some embodiments, te composition docs not contain visible particles when adjudged byhe methods described by European Pharrnacopeia 5 0, Section 220 discussed above [00411 As used herein essentiallyy free of sub-visible particles" refers to a liquid, etg, aqieonis, composition comprising SNS-595 that does not contain subvisible particles 100421 As used herein. "stabWI" refers to a composition comprising SNS-595 that, when stored in a container or vial can remain essentially free of vsible or sub-visible parties or can maintain a specified amount of visible and:or sub-visible particles, for a spccilled 12period ottime. eg. 1, 3, 6 or 9 months. For example. an aqueous composition that is essentially free o visible particles and stable for 6 months with respect to visible pardles refers to an apueous composition that is judged to be essentially free of visible particles at any point in the time during the period starting from a referenced staring point. e,g., the time when the composition was produced and/or added to a container) to 6 months after the composition was prepared. In addition, if the stable composition s a bulk composition, then thatcomposition is capable of being distriauted into a lot of containers, e g., vials, wherein any container containing the distributed composition that is essentiailly free of visible particles at the tine of production of the lo1 is capable of remaining assent of visible particles for the specified period of time eg. 1g 3, 6 or 9 months. in some embodiments, the composition is stable for at least ! 2. 4. 6. , 10, 12, 15, 20. or 25 days In some e1bodiments.the composition is stable for at least , 3. 6 9, 12, 1$8 24. 36, 42, or 48 months. in some embodiments, the composition is stable starting from the time of prodction. [00431 As used herein "time of production refers to the point in tnie when the SNS% 595 conmosinon or cont ainer comprising SNS-595 is produced, nsome embodiments, "tine of productions ins the time when the desired amounts of substantially pure 5NS 595 substance sorbitol water and iethanesulfonic acid are tmixed. In other embodiments the "time of production" is the time when the desired amounts of substantialy pure SNS-595 substance sorbito water, and nethanestlfonic acid are mixed and added to a container c.g. vial. In some embodiments, the time is less than 7 days 6 days, 5 days, 4 daxys 3 days, 2 days, or I day after the composition or contains SNS-595 is produced, In some embodiments the time isless than. 20, 16 12. 8, 4, or 2 hours after the composition or container comprising NS-595 is produced. 100441 As used herein and unless otherwise indicated, the. term harmaceuticaily acceptable salt" include\ but is not limited to. a salt of an acidic or basic group that can be present in the cmpotndR provided herein. Under certainacidic conditions, the cornpound can form a wide variy of saNl with various inorganic and organs acids The acids that cao beu used to prepare phmnnaceutically aceptible salts of such basic compounds are those that form salts comprising pharmacologically acceptable anions include ng, but not limited to, acetate, benzenesulfonate. benzoate bicarbonate, bitartrate, bromide, calcium edetate, 13camsylate, carbonate, chloride bromide, iodide, citrate dihydrochloride, edetate, edisylate. estolate, esylate fumarate; glcetate gluconate, glutamate; g'lycnlliarsanilate, hexyiresorciate hdrabamin' hydroxynaphthoie, isethinnate lactate, lactobionate, malate, maleate mandate, mehanestilfonate (mesylate metiylsulfate, muscate. napsylate, nia pantothente phosphate/diphosphate polygaiactronate, salicylate stearate, succinate, sulatet e a tartrate eelate, triethiodie and pamoateU nder certain asic conditions the compound can form base saks with various pharmacoiogicaliy acptable canons. Noniiting examples of such salts include alkali metal or alkaline earth metal salts and particularly; ca, n; magnesinin, sodium lithium, zinc potassim, and iron salts. [00451 As used herein reprocessing refers to subiecting the SNS-59 (typically obtained fon hsaponicaon Of Con pound 3) o saponification conditions a second ne or more in some embodiments the SNS595 is reprocessed more than one time In some emnbodients the saponification conditions utilize aqueous sodin hydroxide ethanol and acetic acid in some embodments, the saponificadon conditions comprise reacting Compound 3 with an aqueous base followed by netralizing to fbrni. SNS-595 hydrate and dehydrating the SNS-595 hydrate. [00461 As used herein substantially pure Cornpound 3" refers a composition consisting essentially of Compound 3 CO E, MNH N N S Aad V 3, In some embodiments, substantially pure Compound 3 compositions may comprise 0 to 0 3, 0 to 0.25% 0 to 0,% 0 to 0 .1%, to 0,)5%. or 0 to 0,01 iCompound 1, bas'd on total eight of the composition, 14.' Cogi CK N K N N In some enbodimens; ur compound 3 compositions may tOnsiSi essenually of 0 to 03% 0 to 2%, O toO2%, O T 40 005%. orG to 0.01% Compound I, based on tota weight of the conpositon. In som embodmnts substan ialy pu re Compound 3 composnons nmay have 0 to 0 3 o 0 to 025 Oto 0.2% 0 to 0. 1It. 0 to 0 05% 0 r 0 to 0 0 Compound 1, based on total weigh of the copositiOn. In some enbodiments, such caompositios comprise 0 to 0, 1%, or 0 to 0,05%, Conpound based on total weight of th- cnapositioni [00471 The purity of substantially pure SNS95 substance or substantially pure Compound 3 p 4rovidd herein, as well as the amounts of other compounds mentioned herein, can be determined by standard methods of analysis sed by hose (o orinary sk-i in the art sucha - id chromatography (HPLC In tis applicaion reference to a competition having " of any component nans that at least the meacurd amount of that compon ient i ow-r than the limit of detection using such anatc methods or is 0.001 w Or Iess. 4.2 METHODS OF PREPARATION 10048] international Patent Application No. WO 2007/146335 and US Provisional Application No. 61 141856 describe exeamnlary processes known in the art for the synthesis of the SNS 595 compound, As shown in Sheen 2, this synthesis proceeds via IntermiateI which is reacted Ait( Copound 2 in the presence of tretylamne or N N diisopropylethvlarmine to form Compound 3 '. ester group of Conpound 3 is subsequntly hydrolyzed to afford SNS-595 Scheme S to I. b"Can, M a N-' N" ' N S's~ v f~C' < ------------------------------- =J L .) [00491 Bowever, when this process is followed, several impurities can be formed. Specifically, as shown in Scheme 3. following saponification of Compound 3 in basic conditions, approxinnaiely )04% ofi Compomund 4 and. <,5% of'ompound 5 can he observed in the reaction product, in addition to SNS-595. N NN i00501 W thout belingimited to any theory Corpound 5 rnay be' caused by the presence of Compotnd I during the saponification step, indeed, as shown in Scheme 4. the product isolated from the reaction of Compounds I and. 2 in the presence of trietliamine or NN-disopropylethyiamne can contain approximately 0~.3-0.6%? by weight residual Conmound 1 316- Scheme 4 Cl N N compound 2 NE 3or "PNEt IO C C MeHN + CN N NN MeOi 3 1 residua~ (02 3 0,6%) NaCH AcCH EtCH COH C kiH., N N.uO~ N H f' -' N. j l, NN s CL ~O CO2 N''N MeO NNS s N SN5T 4 -004% 0 0 051 Withotein boud to any theory. Cornpoumd 4 may he the result of a reaction between Compound I an hydroxide, possibly through a I A'-addition of water Indeed. when Compound I is treated wih aqueous sodi un hdroxde Compound 4 can hie obtained. in addition to Compound 5, illustrated in Scherme 5 17~' Scheme 5

A

2 Et H CHO- O .. Nc'4N' NA O +H( C N N 04 N N Si N & .4 4.2(a) Reprocessing Method [00521 In ceda i odimets. poss are provided fit preparing substantadv pure SNS-59§ substance by reprocssi a SNS-595 substance. [00531 As noted Th reaction of Conpound 1 with Compound 2 can result in the formation of a mixture of Compound 3 and residual Compound 1. Subsequent t reatment of this mixture with dqueOUs b N sapontation condition can result in the formation of a mixture of SN-595; Compou'nd 4 and Cornipound 5; However when fti mixture is agOin subjected to the aponfcaon conditions, ,e reprocssed the resulting product can contain SNS 95 wt lesser amounts of Coinpound 4 as shown in Sclheme 6. Schemeo 6 - COEt Cr N N N S 1 compound 2 NEt. or trzNEt O 0 0 NC.Et COEt MeHNN N C 'N N N S N S residual (9) 6%) AcOH EtOH OH 0 CHO 32 MeHNt- < ( C N N O c N 2 N S MedO / N 8 N S 04%005% NaOH AcOH EtOH H s.'. &0OH -N N 8 N N NMerHN-"i c- "N' "N' NO c.K N N w6 N'l~ Wls SNS-595 .4 94- Additional experiments show that several cycles of such reprocessing may be used to sequentially further reduce the residual amount of Compound 4 in the composition , In some embodIments. the reprocessing is performed a number of times sfei.cent to reduce the level of Compound 4 to below the limit of detection by conventional methods, such as those disclosed hereim or in he art. [00541 In certain embodiments, reprocessing is accomplished by solubiizing SNS-595 hydrate with an aqueous base fot towed by neutralnizig with acid, followed by dehydration. 100551 in certain embodinments. provided herein are processUs for preparing sibstantially pure SNS-595 substance composing: (a) reacting Compound 3 C veNN N'N N S MeC 3 with a first aqueous base followed by neutralizing to obtain a primary SNS 595 hydrate; (b dehydrating the primary SNS-595 hydrate fro step (a) and reacting the dehydrated product with a second aqueous base followed by neutraidig to obtain a secondary SNS5-395 hydrate; and (c) dehydrating the secondary SS95 hydrate obtained in step (b)to obtain substantially pure SNS-595 substance [00561 The first and second aqueous bases may be the same or different. In some embodments, the first and second aqueous bases of? steps (a) and (l) are each independentypotass iun hydroxide, sodium hydroxide, or lithium hydroxide In some emnbodiment the first and second aqueous bases of steps (a) and (b) are each. independently, sodium hydroxide or lithium hydroxide. In some embodiments, the first and second aqueous bases are each sodium hydroxide. In some embodimentsthe fst or second aqueus base each independently, has a molar concentration of about 0 i- I A M. In some embodiments the first or second aqueous base is an aqueous solution of a hydroxide. -20wherein the first or second aqueous base each independently has a molar concentration of about 0.6. 0.76 0.8, 0.9, 1.09 or abo 1.1 M 100571 The dehydration steps, independently, may be performed by treatment with dehydration solvents known to those of ordinary skill in the art. For example, dehydration may be acconphsned by treating the material to be dehydrated with a oscopic and/or protic sobvent, Exemplary solvents include without limitation ethanol methanol isopropano. and acetone. in some embodensthe Oehydra tonsoivent is anhydrous. 100581 in some enbodinients; the SN-595 hydrate of step (b) or (e Iis dehydrated with ethanol tnome embodinents the ethanol is anhydrous< In particular embodiments, the anhydrous ethanol comprises less than 0.5% water, In some embodiments, the secondary SNS-595 hydrate of step () is dehydrated with anhydrous ethanol at a temperature of about 25-80 *C about 4040 C about 600 N, or about 80 W 100591 Neutralization steps independently, may be accomplished with any acid known to those of ordinary skil in the art. inorganic acids organic acids, or combinations thereof may be utilized. Acids may also he aqueous. Exemplary acids include, without limtation. acetic aid hydrochloric acid, hydrobronic acid, sulfuric acid citric acid, carbonic acids phosphoric acid, oxalic acid, or nitric acid. In some enmbodnents, neutralization is accomplihed with acetic acid. In some emnbodimnents, the neutral izationi step adj usts the pH to 6.0 to 80 I certain embodiments; pH is adjusted to 70 to 8.0. In certain enmbodiments. pH is adjusted to about .3 to 7. 00601 In certain embodiments, the first and second aqueous bases of steps (a) nd (b) are each sodium hydroxide; pH is adjusted to about 73 to 7.7 with acetic acid during neutralization; stepsta) and (b) are performed in the presence of ethanol; an.d dehydration steps are accomplished with anhydrous ethanol at a temperature between about 25-80 "C, t00611 The aqueous base reaction and neutralization may be perbrned in the presence of organic solvents known to those of ordinary skill in the art in certain embodiments the solvent is capable of dissolving SNS595-hdrate when treated with aqueous base and, at the same time capable of pecipitatng NS-595 hydrate alter subsequent neuntrazation. In. some embodiments stp (a) ot step (b) is performed in the presence of ethanol or methanol In some embodiments, step (a) or step (b) is performed in the presence of ethanol It some embodiments, step (a) and step (b) are each performed in the presence of ethanol. The -21volume of ethanol or methanol suitable to carry out steps (a) or (b) may be readily determined by those ot ordinary skiin the art in certain embodiments, step (a)or step (b) is perfoned in the presence of ethanol or methanol wherein the aqueous base is about 1 to about 20% ethanol or methanol by volume, In particular embodiments, the aqueous base is about 3, 5, 0 or 15% ethaiol by voume [00621 In some embodiments, the substantialy pure SNS595 substanceobtaied from step comnprises about 0 to 0.03% Compound 4 based on totalweight of substantially pure SNS-595 substance, In some embodunents, the substantially pure SNS-595 substance obtained from Atep (c) comprises about 0 to 0.02% Compound 4 based on total weight of substantiaWy pure SNS-595 substance. i some embodinents, the substanti pure SNS 595 substance obtained from step (c) comprises about 0 to 0,01% Compound 4 based on total weight of. substantially pure SNS-595 substance. in some embodiments, the substantially pure SNS-595 substance obtained from step (c) comprises about 0 to O.03%$ to 0 02% or 0 to 0.01% Compound 4 based on total. weight of substantially piure SNS-595 substance. [00631 Compound 3 can be obtained Uy the reaction of Compound 1. C' (Ilk N N with Compound 2 MeQ NHMe 2 3TsCH N [(I64j Compound I can be prepared or obtained byany source or method deemed suitable by those of ordinary skill in the art Exemplary methods are described in WO 20 14635 incorporated herein by retrence in its entirety, [00651 in some embodiments; Compound 3 is present as a mixture that comprises about 0 to 0V',3% of Cormpound I based on total weight of the mixture. In some embodininis -22- Compound 3 is present as a mixture that comprises about 0 to 0.6% of Compound 1, based on total weigh of the mixture. In some embodiments, Compound. 3 is present as a mixture that comprises about 0 to 0.3% of Compound 1 based on total weight of the mixture. n some embodiments, Compound 3 is present as a mixture that has 0 to 0.3%, 0 to 0 .%, or 0 to 0 3% Compound 1 based ons total weight of he mixture. In some embodiments, Compound 3 is present as a mixture that comprises 0 to 07%, 0 to 06%, or 0 to 0.3% Compound I based on total weight of the mixture. In some embodiments, Compound 3 is present as a mixture that consists essentiady of 0 to 0 to 0,6% or 0 to 03% Compound based on total weight of the mixture, 4 2(b) Wet NNflisopropylethylamine Method [00661 Inb certain embodiments, Compound 1 ca be reacted witi Compound 2 in the p oN dsoprop y ehylanine(DIPEA) and warin ace itrile to obtain substantially pure Compound 3 which can be subsequently reacted with an aquecous base to obtain substantially pure SNS595 substance. [00671 When Compound I is reacted with Compound 2 in the presence of DIPEA and water in acetonitriicit can be observed that Compound 3 my be obtakied with lower levels of Compound I impurity compared to when the reaction is done in the absence of water As shown in Scheme 7, subsequentaponitaton with aqueous base can provde SNS-95 having lower level of both Compound 4 and Compound 5 compared to reaction of Compound I with Compound 2 in DI PEA in the absence of waten.

Scheme: / 0 %N N NXS compound 2 Pr 2 NEt water M-H N N N SNS 31 0 K 10,II % !saponlificatior M.HN + C N' N SNS-59545 00681 Insome emnbodimentsC u 3 G EI MeHN-/''N N jN Me<0 is prepaed bj racpingfCmpoum4 0 OH2A C.. t , N N wit Compound. 2 MeO, NHMe 02 TsOH, H 2 in DIPEA and water to obtain Compound 3. 100691 Inu soye einbdii'nts, 0.5% to J.0% water in Inetonitrile is used in some cmbodient.%. to 8% Water in acetonitrle is used In sone mbodients 4% to 6% water in acetonitrile is used, [04701 In some enbodiments, water can be added at the beginning during or at thleenid of the reaction, 0071] In some emubodinznts, the process is performed at about 25 'C. In sole enibodiments the reaction mixture may be heatd to effect consumption of Compound 1, in some mbodiments the procs is performed i y a a temperature of about 25 'C and subsequently ataa higher tempature. I some embodiments, t hig hrtemperature is below relu xtemperaure. In a particular embo ditmt the temperature at the start of the reach tion is 25 C s'ch temperature is maintained for a desied ine after which the temperature is raied to about 40-0 C and matitained at about 40 412C for a second desired period. hn further embodiments, the process is performed initially at tmerature of about 25 KC for about 12 hours h) and subsequently at a temperature of about 40-45 "C for about 3-5 hr 100721 In some embodiments, the substantially pure Compound 3 obtained comprises about 0 to 0. about 0 to 0:05%. or about 0 to 03% Commpound -25- .N N s-" N based on total weight of substantially pure Conipoumd 3. In some emibodiments. the substantially pu Compound 3 obtained comprises 0 to 0 1%,0 to 0.05% or 0 to 0.01% Compound 1. [0073I isome embodiments, provided herein are pces preparing substantialy pure SN%-595 substance composing: (a) reacting Conipound 1 C'sa s A K Qt -A N N" c.N veith Compotmd 2 MaQ NHMe 2 T0"' N H 2 in DIPEA and water to obtain substantially pure Compound 3 N N MeO 3 and (b) reacting the substantnaIy p re Cormpound 3 i th an, aqueous base followed by nieutralizing to obtain arnary SNS3595 hydrate; and -26- (e) dehydrating the primary SNS-595 hydrate obtained in step (a) to obtain the substantially pure SNS-595 substance. [00741 In some ernbodinents. step (a) is performed in the presence of actitrj Ie, 00751 some ebodimens, 0,%50 10%water in acetonitrile is used, lasome emi odniN. 2% to 8%waterin acetonitrile is used, Insome emnodimnts. 4% to 6% water in acetone is used. [00761 In some embodiments, water can be added at the beginning, during, or at the end of the reaction, [00771 n some embodiments, the process is perfbmed at about 25 *C, in some embodiments. the reaction mixture may be heated to effect consumption of Compound 1. In some embodiments, the process is performed initially at a temperature of about 25 *C and subsequendy at a higher temperature. in some embodiments, the higher temperature is below reflux temperature. In a particular embodiment, the temperature at the start of the reaction is 25/. s temperature is maintained Qr a despite ime . after which the temperature is raised to about 40-50 C(7 and maintained at about 40-504C fo0tr a second desiid peiod In some embodiments, step (a) is perfTxn'ed tnitialy at about25 'C and subsequently at about 4045 1 In a particular embodiment, the step (a) is perfrned nitally at about 25 7 for about 1,2 hr and subsequently at about 40-45 C ifr about 3-5 hr, In some embodiments the step (a) is performed at about 25 *C. [fO78] In some embodiments, the substantialy pure Compound 3 obtained comprises about 0 to 0. 1%, about 0 to 0.05%. or about 0 to 0,03% Compound I COE based on totahweight of the substantially pure Compound 3, In some embodiments, the substantially pure Compound 3 obtained comnprises 0 to 0. 1%, 0 to 0.05%. or 0 to 0,0 Compound 1. 1(10791 In some embodiments, the aqueous base is potassin hydroxide, sodium hydroxide, or lithium. hydroxide, In some embodiments,the aqueous bse issodiu hydroxide or lithium hydroxide, in some embodiments, the aqueous base is sodium hydroxide. In some emnbodiments, the aqueous base has a mnolar concentration ot about 06 IA MI In , some embodiments, the aqueous base is an aqueous solution of a hydroxide having a molar concentration of about 0.6, 0 ,08 0.9, 1.0 or about I M, [00801 In some embodiments, step (b) is performed in the presence of ethanoI or methanol. In some embodiments, step (bi is performed in the presence of ethanol The volume of ethanol or methanol suitable to carry out step (b) may be readily deternined by those of ordinary skill in the art In certairembodiments, step (b) is performed in the presence of ethanol or methanol, wherein the aqueous base is about I to about 20% ethanol or methanol by volume, in particular embodiments. the aqueous base is about 3, 5, 10. or bout 15% ethanol by volume. ff0811 In some embodiments, an. acid is added in step (b) after reacting Compound 3 with the aqueous base to adjust the pH to 6.O to 8.0. Suitable acids include, but are not limited to acetic acid, hydrochloric acid, sulfuric acid and the like i certain embodiments. the acid is acetic acid, In certain embodiments, pH is adjusted to about 7.3 to 7,7. [00821 In certain eibodimients, the aqueous base of step (b) is sodium hydroxide;r 1 is adjusted to about 7i3 to 7.7 with acetic acid after Compound 3 is reacted with the sodium hydroxide in step (b and step (b is performed in the presence of ethanol. [00831 In some embodiments, the primary SNS-595 hydrate f step (c is dehydrated with ethanol in sonic embodiments, the ethanol is anhydrous n particular embodiments the anbyvdratus ethanol comprises less than 0.5% water. In some embodiments, the primary SN S-595 hydrate of step (c) is dehydrated with anhydrous ethanol at a temperature of about 25480 "W, about 40-80 'C, about 60-80 or about 80'C, [00841 In certain embodiments, the substantially pure SNS-595 substance obtained from step (b) comprises about 0 to 0.02% or about 0 to 0.1% Compound 4 OH 0 C1 N' S N 4 -28based on totl weight of substan tially pure SNS-595 ubstanece. In some enbodiments. ,the substantially pure SN595 substance obtained frome (b)has 0 to OQ02%O r 0 to 0,01% Compound 4. In some embodiments, the substantialy pure SNS-595 substance obtained from; step (b) consists essentially of 0 to 0.02%,or 0 to 0.01% Compound 4. In some ernbodirentsthe substantially pure SNS-595 substance obtained from step (b) comprises 0 to 002%or 0 to P1% Compound 4, [0086I In some embodiments the substantially pure SNS-59 5 substance obtained. from step (b) comprises about 0 to 002% Compound 5 0 0 'OH Cl N' N S N based on total weight of substantialy pure SNS-$95 substance, Insomne embodiments, the substantially pure SNS-595 substance obtained from step (b) has 0 to 0,02% Compound 5. Income enbodments, the substanially pure SNS-595 substance obtained fom step (b) consists essentially of 0 to 0.02% Compound 5. In some embodiments, the substantially pure SNS-595 substance obtained from step(b) comprises 0 to 0.02% Compound 5. 100861 In some embodiments. the substantially pure SNS-595 substance obtained in step (b) is reprocessed in some enhodm ents, provided herein are processes for preparing substantially pure SNS-595 substane comnprising (a) reacting Compound I with Compoondt 2 -29- MeQO NHMe *2TsOH H 2 in the presence of DPEA and water to obtain substantial pure Compound 3 MeHN AON S 3 (b) reacting substantiay pure Connound3 with a first aqueous base followed by neutralizing to obtain a primary SNS-595 h\yrate (e) dehydralin te primary SNS-595 hydrate obtained in stp (b) to ftrm a SNS-95 substance; (d) reacting he SNS-$95 substance om step (c) with a second aqueous base followed by neutraHzing to tain a seondary SNS -595 hydrate; and (e) dehydrating the sceondar SNS195 hydrate obtained in step() to obtain substantially pure SNS-595 substance 4.3 COMPOSITIONS [00871 Compostions are provided that comprise substantially pue SNS'9-55 substance, Compositions are also provided that consist essentialy of subsamtially pure SNS-595 substance. Compositions are aso provided that consist of substantially pure SNS-595 substance. [00881 In somew embodiments, conpositIons are provided vherein te substantialy pure SNS-595 substance comprises about 0 to 003% Compound 4 OH C 06 N'O 0 7 &N -30based on totl weight of substantially pure SNS-595 ubstance. In some embodiments, the substantially pure SNS-59 substance comprises about 0 to 0,02% or about 0 to 0.01% Compound 4. In some embodiments, the sostantially pure SN595 substance eompnses 0 to 0.03% 0 to 0.02%. or 0 to 0 %01 Compound 4. [OOS9] In some embodiments, substantially:pure SNS59 substances are provided that consist essentially of SN S55 and about 0.03 wtk% or less, about 002 wt%or less, or about 001 w% or less Compound 4, In some embodimentsthe coiposiion consists essentially of SNS-595 and 003 w or less, 0;02 wtor less, or 001 wt% or less Compoumd 4, In some embodiments, the composition consists of SNS-595 and 0.03 wi% or less, 0.02 wt% or less or 0.1 wt% or less Compound 4, [00901 In some embodiments, the substantially pure SNS 95 substance comes about 0 to 0.03%, about 0 to 0.2%,or about 0 to 0,01% Compound 4. at the time of production. In some embodiments, the conposition comprises 0 to 0.03% 0 to 0102% or 0 to 001% Conpound 4 at the time of production [0091] in some embodiments; substantially pure SNS-$95 substances are provided that consistssta~y of SNS~595out 022 wt% or lessor about 0.01 wt% or less Compound 4. at the time of production. In some embodiments, the composing consists essentialy of SNS-595 and 0.03 wt% or less, 0.02 wO% or less, or 0.1 wt% or ess Compound 4, at the time of production. Income enmbodients the composition consists of SNS095 and .3 wt% or less, 0.02 wt% or less Compound 44 at the time ot production. 100921 In some en bodimerts, composhions are provided wherein the substantially pure SNS~595 substance comprses about 0 to 0.04% Compound 5 0 0 N' N based on total weight of substantially pure SNS-595 substance in some embodiments, the substantially pure SNS-595 substance comprises about 0 to 0.03%, or about 0 to 0.02% 31- Compound 5. In some embodiments, the substantially pure SNS-595 substance comprises 0 to 0.04%. 0 to 0,03%. or 0 to 0 02% Compouind 5, {00931 In some embodiment substantial pure SNS595 substances are provided that consist essCntially of SNS-595 and about 0,04 wt.% or less, about 0,03 wt%or less, or about 0.02 wt or less Compound 5. In some emboimens, he compositon consists essentially ofSNS-595 and 0.04 wt% or less, O.03 wt% or lesso 0.02 wt% or less Conpound 5. In some embodiment te composiion consists of SNS-595 and 0.04 wt% or less, 0.03 wt% or less. or 002 wt% or less Copound 5, [0094 nsome eimb imts, the substantially pur SNS-595 substance comprses about 0 to 0,04%. about 0 to 0.03%, or about 0 to 002% Compound 5, at the time of production. In some embodiments the composition comprises 0 to 0.04%.0 to 0.03% or 0 to002% Compound 5 at the time of production. 100951 In some m e he substantially pure SNS-595 substance consists cssen.ialV of SNS-595 and about 0.04 wt% or less, about 0.03 wt% or less, or about 0.02 wt% or less Compound 5, at the time of production. In some enibodimentsthe composition consists essentialyof SNS-595 and 0.04 wt% or less, 0.03 wt% or less, or 0.02 wt% or less Compound 5, at the time of production In son enbodiments. the composition consists of SNS095 and 0.04 wt% or less;0.03 wt% or less, or 0.02 wtor less Compound 5, he time of production. [0096] in some embodiments, substanti ally pmre SNS-595 substances are provided. wherein the substantial ly ptu SNS-595 substance comprises about 0 to 0.07% total Compound 4 and Compound 5 combined based on total weight of the compositions. In some embodimentshe composition comprises about 0 to 0,05% total Compound 4 and Compound 5 combined In some embodiments, the substantial composition comprises about 0 to 0,03% total Compound 4 and Compound 5 combined. In some embodunents, the composition comprises 0 to 0.07%., 0 to 0105%, or 0 to 0.03% total Compound 4 and Compound 5 combined. [0097] In some embodiments, substantallypure SNS-595 substances are provided that consist essentially of SNS-595 and about 0.07 w% or less, about 0.05 wt%or lessa or about 0.03 wt% or less total Compound 4 and CoInound 5 combined. In some embodiments the composition consists essentially of SNS 595 and 0.07 wt% or less, 005 wt% or less, or 0.03 -32wt% or less total Compound 4 and Conpound 5 combined In some embodiments, the composton consists of SNS495 and 0,07 wt% or less, 0.05 wt% or less or 0.3 wt% or less total Compound 4 and Compound 5 combed [00981 In some pure SNS-595 substance comprises about 0 to 0.07% about 0 to 0.50%, or about 0 to 0% tota.Comnpom 4 and Compound 5 combined, at the time of production. In some embodiments, the composition comprises 0 to 0.07%, 0 to 0.3%, or 0 to 0'03% total Compound 4 or Compound 5 combined, at the time of production. [00991 I soni embodiments substantially pure SNS5595 substances are provided that consist essentially of SNS-595 and about 0.07 wt% or less about 0.0 wt% or less, or about 0.03 wi% or less total Compound 4 and Compound 5 combined, atte time of production. In some embodiments. the composition consists essentially of SNS595 and 0.07 wt% or less.0 05 wi% or less, or 0.03 wt% or less total Conpound 4 and Compound 5 combined, at the time of production some embodiments, the composition consists of. SNS-595 and .07 w:% or less 0.05 wt% or less, or 0.03 wt% or less total Compound 4 and Compound 5 conbnied at the ime of production. 1001001 The presence of Compound S in a solution comprising SNS95 and water can result in sub-visible microscopic paieses: The presence ot microscopic or subisible particles can be determined by any technique deemed suitable by one of ordinary skill in the art For instance the number of particles can be determined by the obscuration method specied in USP~NF General Chapter 7 8Which is incorporated herein by reference in its entirety and described below. Alematively, flow imaging telmiques (such as that available from Brightwell Technologies, c-nay be used to determine the particulate matter content of a composition. t[0l101} In some embodiments, compositions are provided wherein abou 100 mg of subtanialy pure SS595 substance is present fbr evcry 10 mL of the composition; and wherein the composition has not more than 80. not more than 70, not more than 60, not more tihan 50 not more than 40 not more than 30, not morehan 20, not more than.15, not more than 10, or not nore than 5 particles 25 microns per 10 ni of ther coposition 00102 j In some embodiments, compo sitio ns are provided wherein about 100 mg of SN S-595 is present for every 10 niL of the composition; and wherein the composition has .0,not more than 3000, not more than 2500 not more than 2000 not more than 1500, not more than 1000, not more than 800. not, more than 645. not more than 600, not more than 300, or not more than 100 parties 10 microns per 10 mL of the compositon, 1001031 In some enibodiments, comnositions are provided that comprise SNS5'95 and 0 3 mg or less 0.2 mo or ,or 0.1 m or less Compound 4 per gram of SNSK5IQ. In some embodiments, the co positioWs are aqueous solutions of a substantiallypure SN. 595 substance, eg, 3 mg or less Compound. 4 per 10 gn of the substantially pure SNS-95 substance per 100 niL solution, In some embodiments, the compositions further compose sorbioit In some embodiments. the sortol present in ananount providing a 4. 5% aqueous soludon of sorbitol, In some embodiments, the compositons fiflhe comprise methanesultnic acid In some embodiments, the methanesuhfnic acid is present in an amount to provide the solution a pH of 5. 1001041 In some embodiments the conposition is an aqueous solution consisting essentially' of? (a) 10 g of a substantially pure SNS-595 substance consisting of SNS 595 and Conpound 4, wherein the substantial pure SNS~595 substance conias 03 mg or less Compound 4 per grarm; (b) 45 g of sorbitol: and (c) sufficient methanesulfonic acid to provide a pH of 2.5; per 100 mL. of the solution Also provided are products comprising a container containing aun auot of such solution eg.1.0 m of suc sohtion [00105} A composition comprising SNS-595 and 04mg or less 0.3 mg or less, or 02 mg or less of Compound per gram of SNS-595. In some embodiments, the composition arc aqueous solutions of a substantially pure SNS-595 substance, e g.. 0.4 mg or less of Compound 5 pert gm of thep t a pure SNS495 substance per 100 mL. solution. In some embodiments the compositions further comprise sorbitol In some embodiments the sorbitol is present in an amount providing a 43% aqueous solution of sorbitol. In some ermbodimenusthe compositio.ns further comprise methanesultone acid. 3nsome embodiments, the methanesulfonic acid is present in an amount to provide the solution a plH of 2.5. -34- [001061 nso embodiments the composition is an aqueoussolution consisting essentially of: (a) 10 g of a substantially pure SNS-595 substance consistin.g of SNTS~ 595 and Compound 5wherein the substantially pure SNS-595 substance contns -4 (g or less Compound 5 per grand; (b) 45 g of sorbitol: and (c) sufficient rethanesufwoni acid to provide a pH of 2,5; per .100 ml. of the solution. Also provided are products comprising a eontainercontining an aliquot of such solution, e g.) 0 mL of sucht solution. [001071 A composition comprising SNS%595 and 0,7 mug or less total Compound 4 and Compoumd 5 per gram of SNS-595. A composition comising SNS-595 and 04 mg or less, 0.3 mg or lessor 0.2 mg or less Compound 5 per gram of SNS-595, k some embodinents. the compositions are aqueous solutions of a substantlidy pure SNS-.595 substance eg. 4 mg or less of Compound 5 pen 0 gm of the substanialy pure SNS-595 substance per 100 rmi solution, In some embodiments, the compositions further compose sorbitol In some embodiments, the sorbitol is presn in an amount providing a 4.5% aqueous sohtion of sobitol In some embodimentsthe compositions further compose nmethanesulfonic acid, in some enmbodinrents; the methanesulfonic acid is present i an amount to provide the solution a p- of 2.5, [fO1O8 in some embodiments the composition is an aqueous solution consisting esseutialigy of. (a) 10 g of a substantially pure SNS-595 substance consisting of SNS 595. Cornpoun.d 4, and Corpound 5 wherein the substantially pure SNS-595 substance contains 07 mg or less total Compound 4 and Compound 5 combined per gram; (b) 4,5 g ofsorbitol and (c) sufficient methauesulfonic acid to provide a pH of 2.5; per 100 mi> of te solution. Also provided are products comprising a container containing an aliquot of such solutioneg.10 ml of such solution. -35- [001091 Further, the presence of Compound 4 in a solution comprising SNS-595 and water can resuk in the formaton of vNsible partics. Without being limited to any theory, the visible particles can compriseapound 6 OH OH Ch CA L I 1CC N;N C I S IN N S \=I \= 6, which can be derived from Compound 4 Without being limited to any theoryCompound 4. when exposed to formaldehyde, can react to forn Compound 6, indeed, when Compound 4 treated with formaldehyde. the formation of Compound 6 has been observed, [001101 In some embodiments, presses are providdthatwhen compared to processes known in. the art. produce substantially pure N%~595 substance inchiding lower amounts of Con,,rounds 4 and 5. an,-W th-us lowevr aonsof partichles, e~g,- C'ompound 6, when such substantiaiy pure SNS-595 substance is provided a aqueus solution [00l111 In some embodiments, processes are provided that when compared to processes known in the art, produce substantially pure SNS-595 substance including lower amounts of compound 7 Cad [001121 In soene enibodimernts composions conlprising substantially pure SNS-595 substance are provided wherein the compositions are essentially free of vsibl parties and are stable over tirne in some enbodiments the composition is stable for 1, 2 6 8. 10 12, 15, 20, or 25 day(s) In some embodiments, the composition is stable for 1, 3 6 9 1 2, 18, 24, 36, or 4 monthss, In some embodiments. the composition is stable starting from -36the tite of prodIetior. lit some embodiments the comosition is stable when contacted with fonnal dehyde. In scae embodiments the composition o is stable vhen contacted with a compound capable of transforming Compound 4 OH 0 H S N to Compound 6 OH OH C' h CK N N C) OC N N ^C S 'N N S 6. [001131 As discussed. visible parties may have, in some embodiments. an average diameter of at least 50 pm, at least 75 pm. at least 100 pim, at least 150 pn or at least 200 p. In some embodiments the visible particles have an average diamStOr ofabout 50500 pm, about 50-300 am, about 1.00-500 pnn. or about 100-300 prn. 1001141 The crystallinity and crystalline habit may be determined using methods known to those of ordinary skil in the art. [or example 'rystallinity and crystaihne habit nay be assessed by polarized light microscopy. I some embodiments, the visible parties are crystalline powders. in someenmodiments, the crystalline habit of the visible parties is that of plates about 45 pn to about 150 p1M. or about 50 pm to about 100 4 m, [001151 The detection of visible particles in a composion can be determined by any technique deemed suitable by one of ordinary skill in the art. For instance visible particles may be detected by the method specified in European Pharmacopeia 50 section 2.9.20, which is incorporated herein by referenced in its entirety, Certain exemplary techniques are described ifl grea ter de tat below [001161 i some embodiments, visible particle presence is determined at an illumnation nternsit between about 2000 and 3750 lux. -37- [00117 In some embodiments the visible parties comprise compound 6, [00-181 In son embodiments the visible particles comprise compound 7. f00-191 In some ernbodinnents. processes ae provided that, when compared to processes known in the art, produce substantially pure SNS-595 substance inchmding lower amounts of sub~visible particles as detected by thefLigh t Obscuration Particle Count Test as described i the tS. Pharmacopoeia 88> Particulate Matter in Injections the entirety ofwhich is incorporated herein by reference. In some embodiments; the sub-vsible parties comprise one or more of compounds 6. and/or 7, [001201 isome ebod iments, provided herein are compositions comprising substantially pure SNS4595 substance. wherein the compositions contain not more than 6000 sbvisible particles i un per vial, not more than 3000 sub-vsibk parties 10 pm. per vial, or not more than 1000 suhisible particles 1 0 pm per vial as measured by light obscuration. In some embodiments, provided herein are compositi ons comprising substantially pure SNS-595 substance, wherein the compositions contain not more than 600 sub-visible particles particles R 25 pm per vial, not more than 300 sub-visible particles parties :25 pm per val, or not more than 100 sIubvisibe patices .25 tmi per vial by light obscuration. In some embodiments, provided here in are conmositions comprising substantially pure SNS-595 substance wherein the compositions contain not more Itan 3000 sub-isible particles - 10 pm per vial, not more than 1500 subvisible particles > 10 pvm per vial, or not more than 300 sub-visible particles > 10 am per vial as measured by microscopic evaluation. In some embodiments, provided herein are compositions comprising substantially pure SNS-595 substance wherein the compositions contain not more than 300 sub-isible particles 25 m per vial, not more than 150 sub-visible particls R! 25 tm per vial, or not more than 30sub-isible particles it 25 tpm per vial as measured by microscopic evaluation.. 1001211 In some etnbodiments, he composition comprises sorbitoLI. hifrther mbodi ments the composition comprises methanesulfonic acid, In further embodinents, the sorbitol is present in an amount providing a 4,51% aqueous solution oftsorbitol In some embodiments the mezhancsulfoniC acid is present in an amount to provide a pH of .5, in oe embodiments, the composition is stable. 38~ [001221 insome embodiments, the composition consists essentially of 100 mg substantially pure SNS-595 substance, 450 mg of iwsorbitol, water, and methanesul1onic ac di d herein the meIhaneslfonic acid is present in amount to provide a pH of 2S, and wherein the water is present in an amount to praide a total composition volume of 10 nt In some cmbodiments, the compositon is stable [001231 in some embodiments, the composition consists essentially of water, substantially pure SNS-595 substance, sorbitol, and methanesulfonic acid, n some embodiments, the composition is stable. jI1241 hn some embodiments compositins consisting essentially of SNS-59 and 0.03 wt% or less Compound 4 are provided, OH 0 S; 1N , N 4 1001251 In sorme embodiments, composidons consisting essentially of SNS-595 and 0,04 w% or less Compound 5 are provided 0 0 TOH C I N N S N (001261 In somec emibodiments, compositions consisting essentially of SNS-595 and00 wnor ess toa n mound 4 and. Company 5r rovided~ 0 H 0 0 4a, ~j ( N We"tHN"'" s N S N 4 5. -39- [00127 In somne enbodiments. eonpositiOns comprising SNS§95 and 0,3 mg or less Compound 4 per graim of SNS~595 are provided. OH 0 01 ' N ,?0 SK I'< N \ / 4 [00.128 In some embodim.entscom.npositdons comprising SNS~595 and 0A mg or less Compound 5 per grain of SN8%595 are provided 0 0 OH c I N N S N [1)01291 hi some eimets, composi tons comprising SNS~595 and 0,7 mg or less total Compound 4 and Compound 5 per gram of SNSr95 are provided. H OH s N s N 4 5 1001301 In some: enbodiments compositions consisting essentially of: (a) 10 g of a substanialy pure SNS-595 substance consisting of SNS-595 and Compound 4 wherein the substantially pure SNS595 substance contains 3 ug or ess Comipoi4d 4 per gram OH 0 014 S'N 4 (b) 45 g of sorbitao and (C) sufficient methanesulfo.nic acid to provide pH or 2.5 per 106 mL. of the solution are provided In sonie embodiments a product conpising a container containing 10 mL of such solution is provided. (O01311 In some embodinvents; an aqueous soution consisting essentially of: (a) 10 g of a substantIaly pure SNS-595 substance con ising of SNS 95 and Compound 5 0 0 5 OH or N Nt wherein the substantialNy pure SNS-595 substance contains 4 mg or less Compound 5 per grant (b) 4, g of sorbi tol: and (c) sufficient methanesuhfnic acid to provide a PH of 2 per 100 mitL of the solution is provided, In some enibodimcnts a product comprising a cotiner containing 10 mL of such solution is provided 1001321 In some cmbodnnts an aqueous sohuion consisting essentially of: (a) 10 g of a substantially pure SNS-595 substance consisting of SNS lCom pound 4 and Compound 5 -41- OH 0 0 0 Von An10 cl N FN- N7j NXc S N S N 4 5 whetrein thle substantialy p ure SNS-595 s t c ains 7 mg or esst Compound 4 and Coompound 5 combined per gram; b) 45 g of s5Wbi toand (c) sufficient methanesuifonic acid to provide a pH of 2.5; per 100 mL of the solon is provided. nsome embodiments a product comprising a container containing 30 nmL of such solution is provided, [001331 Also provided herein is a compound of formula 6 OH OH C1 N N 0 C VI N Q S N N S 6 or a pharmaceutically acceptable salt thereof. 100134j Also provided herein is a compound of formula 7 c0wi or aphrmetcalaceabeslthro 10013 51 Cena in em b odiment of the cla0imed. subj.ct m atter are fi ,ustrated by the0 -42- 101 361 As not edhoe reaction of Compound 1 with Compound 2 can eu in the forntim of a rmix ure of Compound 3 and residue Compound I. Subsecqut trarict of this mixturC with aqueous base, L wponifin 00dAonWT lpor e CaqUCo sodim hydrdxide. can result in the frmation of a nixture of SNS-593, Compound 4 and Compound 5 Hoaeverwhen thi\ mixture is again subjected to the saponfication conditions Le, processed, the resihng product can contain SNS-595 with lower atnontinis of Compound 4. as seen above in cheme 6, [V! 71 hxperinents were performed to assess the efectof soh reprocessing of SNS 595 substance on the rsidual levels of Compound 4. The results of these experiences are presented in Tabe 1 Tahk educion ( (>rnou4d byr Compn d Coipound 4 atv'r 4 sp ked eprocafegr (actual Entreatment 2 06 ( None 0 2 4 I09 0,06 None QU6 ND) 3 6 s g 03 NA 001 . 3) 4 7 25 012 NA 0005(00 5 8 25 g 0. NA 002 0 00 6 9 40 g ( 02 NA ( 004) 7 000 g 0 02 NA (0 005 14 Tkg ( 0. N < 91 2 2 NA SNS51 with 0 2% Conpound 4 s used NA = not avail NDb = N) Non detcd A3- [001381 Experinent : To a solution of sodium hydroxid gIin water (5 ml), SNs595 (098g) Compounid 4 (0/010 g) adl ob(EtOH) (1 ml) were added. The mixture was filtered and tv' He PDC analysis of the filtrate showed Compound 4 as about 0 7% (by area) The pH of the filtrate was adjusted 6 by sdow add iton of acetic acid and the solution was heated to precipitat SNS-595 hydrate The slur Iwas cooled and flted to gi' SN-595 hydrate. HPLC analysis of the solid showed it to have 006% (by area) of Compoutind 4, dic ating that a ARM-old reduction in Compound 4 had occurd, ihe solid was resubjected to reprocessing using aqueous sodium hydroxide (0 8 g in 6 ml. water) and EtON (1 mI followed by pH adjustment. with aqueous acetic acid. After heating. the solid was filtered to give SNS-595 hydrate having less than 0 01% Compound 4, demonstrating at least a further sixlold reduction in the amomt of the con taninant 1001391 Ejurjnenth SNS-595 g) was spiked with I% Conmpound 4 HI PLC analysis of tin spiked sample folning eprocessing conditions, t retmnt with NaOH, showed 0i9%K Co'mpound 4 by area This mate rawas then p dusted and the solid isolated as above, HPLC analysis showed 0 06% Comnpound 4 by area, indicating that a 40fold reduction i Coipound 4 occurred onsistent with the frst expernmet aboe. This material, when. again subjected to reprocessing co ndiions,provided a SNS-595 substance wit no detectable Compound 4. demonstrating at least a sixfbld reduction in the amount of that contaminant; 100140J ScaLb _Eip its Eperiments 3 9 Were performed to evaluate the scalability of this process. The procedure for Experment 8 is provided as an exemplary procedure R.esuls for Experiments 3-are sumize~td below and o Tabl 1 [00 1411 Exeruay Procedure for atge Sca> ReprocessiRg Erxperment 8)k To a sotion of NOH (092 kg) in water (4.8 kg S - 1N595 (4 kg}having Compound 4 (~ 24% by weight was added 'To the tnixfure" EOH1 (013 kg) was added. The solution was fStered and the pH of the ftrate was adjusted to 73.7y slow addition of aqueous aretic acid. The mixture was heated to 55- 05C cooled and filtered Th fit cake was washed with water, EilH and dried under vacum to give a SNS&595 hydrate (1.36 kg) having. 0,013% by weight) Compound 4. The hydrate (1,36 kg) was dehydrated by -44scurrying in DOH (23 kg) at 67-78 ' Afer cooling, the mixture was filtered, washed with EiOH and dried at 65-75 Cl under vacuum to give a SNS-595 substance (1.0 kg) having icss than 0.005% Compound 4. 1001421 M S man of Scal li >;e iy En rrments [001431 In Experiment 3, 25 g of SNS~595 having 0.3% Compound 4 was sthjected to reprocessing conditions, ie sodium hydroxide. followed by pH ajstmentand dehydration. PLC analysis of resulting SNS-595 showed 0 a1% Coipound 4 by area, indicating that a 30 fold reduction in Compound 4 had occurred. [001 44j in Experiment 4, 25 g of SNS~5% having 02% Caopound 4 was subjected to reprocessing conditions, Le,, sodium hydroxide foIllowed by pH adjustment and dehydration HPLC analysis of resting SNS-595 showed 0,005% Compound 4 by area, indicating that a -40 fold reduction in Compound 4 had occurred. 1001451 In Experiment 5, 25 g of SN S-95 having 0 1% Compound 4 was subjected to processing conditions ie, sodium hydroxide, fllo wed by pH adjustment and dehydration, HPLC analysis of resulting SN-S55 showed 0. 002% Compound 4 by area, indicating that a fold reduction Compound 4 had occurred. 1001461 In Experiment 6, 40 g of SNS-595 having 0.02% Compound 4 was subjected to reprocess condetionse sodium hydroxide followed by pH djuuenan dehydration. HPLC analysis of resulting SNS-595S showed 0.004% Compound 4 by area, indicating that a -5-fold reduction in Compound 4 had occurred. 1001471 In Expriment 7, 200 g of SNS-595 having 002% Compound 4 was subjected to reprocessing conditions, i sodium hydroxide, followed by p1-I adjustment and dehydration. HPII. analysis of resulting SNS-595 showed less than 0.005% Com pound 4 by area, indicating that a 4-fold reduction in Compound 4 had occurred. [00 1481 In. Experiment 8, 1 49 kg of SNS-595 hang 0.02% Compound 4 was subjected to reprocessing conditions, ".sodium hydroxide, followed by pH adjustment, HiPLC analysis showed < 0005%4 Compound 4 by area, indicating that a -4 fbld reduction in Compound 4 occurred [001491 in Expriment 9, 12 kg of SN15-5.95 having 0.02% Compound 4 was subjected to reprocessing conditions. i e sodiun hydroxide, followed by pl adjustment and -45'dehydration HPLC analysis of resulting StNSJ95 showed 0:05% Compound 4 by area, indicating that a>24fold reduction in Compound 4 bad occurred, Example 2i Reduction of amount of Com found 6 1001501 As discussed, the presence of Compound 4 in a SNS-595 solution may be attended by the formnation of visible particles which, without being limited to any theory, compose Compound 6. Experimentsere performed to assess the effect of reducing Compound 4 levels on the formation of Compound 6 over time. (001511 Four different sample drug product solutions were prepared from the drug substances havig. varying I evls of Compound 4 outlined, in ble 2, below, Sample used the repimeessed product obtained fromEperiment 6 (described in Example 1) and had <0.005% Compound 4, Whenever extend addition of Compound 4 was made. Compound 4 was first dissolved in a 5 mM aqueous NaOH- sohnion A total of 2% of the aqueous NaMH solution was added to each of the drug product Samples for consistency purposes [001521 The various drug product Samles were filered twice through a 0.22 pm PVDF SieriCup fiIters. These pre-autoclaved, filtered drug pmduct Samples were analyzed for their Compound 4 content six days fW low prepan'ation. and storageat room temperature (R') shielded from light. Figure 1 and Table 2 show that the amount of Compound 6 in the drug product correlated directly with the level of Compound 4 in the SNS-595 drug substance, The filtered dmg product Samples were then filled in 30 nL Schott glass vials and stoppered Vial fiing and stoppering were performed in a laminar flov hood, Vials and stoppers used in this study were rinsed with filtered water and allowed to dry in the laminar flow hood before use. Vials were not depyrtgenated and stoppers were not sterilized before use, Filled and stoppered vials were crimspcaled, autoclaved, and stored upright and shielded from light at 40 '/75% native humidity (RHin After 65 days of storage the vials were inspected for the presence of visible particles and analyzed by HPLC for Compound 6 content Analysis of the Samples having less than 0 01% oonpound 4 (entry 1) showed that the resulting drug product had no visible particles and no detectable amount of Compound 6 after 65 days. Experiments with Sampe having L0:02% Compound 4 showed signfcant formation of Compound 6 and visible particles during the sae time perod (Samples 2-41 The amount of Compound 6 after 65 days correlatd directly with the nidai amount of Compound 4 in the SNS-595 substance Saple Nomnin 4 Compound 6 Copound 6 Coinp(mnd 4 in (,i gal) ( Qg/va. ) Dlg Substance After - day After 65 days 2 0 . 0. .9 .3 0 04 214 V9 ___I___ O) -9 16 -- 4 Exapk3: edcton f ompuns an 5wvia Wet DPLA [001531 Experiments were performed to assess the effect of water on the reaction between Compound 1 and Compound 2 in the presence of DIPEA. Experiments were also performed to comparethe effect of adding water duing the beginning of the reaction with adding water at the end of the reaction. [001541 When Compound I and Compound 2 were reacted at 40-45 C in the absence of water analysis of the product showed between 026% and 03.1% residual Compound I by area, [001551 In a second series of experiments Compound I and Conpound 2 were reacted at RT? for 1.2 hr. and subsequently heated to 40-45* C fbr 3-5 hr. In these experiments water was added at the beginning of the reaction Analysis of the product showed between 0,0 % and 008% residual Compound I by area 1001561 In a third series of expimns Compound I and Compound 2 were reacted at RT for 12 br, and subsequently heated ro 45 C for 3 hr. In these expeiments water was added at the end of the reaction. Analysis of the product showed 0.03% to 0.09% residual Compound i by area The experiments above are sumnmarized in the table below -47 Tbe3Ecof in SNS 595sy oe _ Rxn Conditon Water Scale Residua Cominents (volume) Comound DIPEA 40 5 "C 50 g 026 S031 No water DIPFA, RTd 12 hr, 1 0.05 Water added at then ated to 40- 1 0 0 08 the beginning 50 000 4z C(. 3-5 hr 0,5 5o00 DPEA RT% 12 hr 1 g 0.03 \ Water added then. eated o 40- 05 0 g 005 at er 12 hr at 45 'c3 hr 50g 009 Anyzed at 275 nm1 [00157I The following is an. exenplary procedure for the experiments desenbed in the Table 3 above. (00158 To a sluny of Compound 2 (1.55 kg) in acetonitrile (ACN: 10 LV DIPEA (4 L) and water (0.5 L were added. To the sohition. Compound I (1 kg) and ACN (1 ) were added and the reaction was starred tor about 12 hr at RT Te reaction mixture was then heated to about 45 C for about 2-6 hr, After cooling, the product was filtered, washed with ACEN(4 14 and dried under vacuum o give Compound 3 (1. kg PLC analysts showed this materid to eontin< 0, 1% Compound 1, Experimens were cared out to determine the i4opact of residual Compound .1 levels on tei Compound 4 levels resulting rng the manufuetre of SNS-595. These experiments demonstrated a elation between Compound I levels in Compound 3 and Compound 4 levels in the resulting SNS-59 substance. Experiments at laboratory scale also denonstrated that Using Compound 3 substance having <0 01% Compound I provided a SNS-595 substance with 0.01% Compound 4. he results of these experiences are summarized below in Table 4 and Figure 2. -48 Table 4: I-npac; frs&v Co alpoitl 1, i Compound "3 on Compound 4devel in SN% 59 .try Sca. Compound. Compound 4 1-' tr , ;v 1015 kg A 0 04 2 0,75 kg 0 60 0,05 3 21 kg 0.64 0.04 4 5 kg 0.24 0,02 0 0.8 0,00T 6 50 g 009 04008 7 1 kg 0.01 < 005 1001591 The o] lowing is an exempar procedure for the experiments described in. Table 4 above. [001601 CoimpoInd 3 (0.9 ky havingg 0.1% Compound 1.) was added to a solution of sodium hydroxide (014 kg) in water 3 kg) and EtOH (0.16 L) The mixture was stiixed fOr about 12 hr and filtered to remove insoluble materials. The pH of the filtate was adjusted to 7.3-.7 with aqueous acetic acid, The resulting mixture was heated to about 60 C for24 hr cooled and fi.tered The lter cake was washed wh water, EtOH and dried under vacuum The resuting SNS-595 hydrate was shirried in. EtCH (20 L) at about 70 'C for about 4 hr, cooled and fitered, The filter cake was washed with [sOb and dried at 55 75 W under vacuum to give SN&-595 substance (0.66 kg) hayingless than 0. 00% Compound 4 (Table I entrv ;) [001611 in a sumiAr manner, experiments were carried out to determine the impact of residual Compound 1 levels on the Compound 5 levels in SNS-595. These experiments dem nstrated a correlation between Conpound I levels in Compound 3 and Compound 5 levels in SNS59$5 The resuUs of these experiments are summarized below in Table 5 and Figure 3. -49able 5: Impact of residual Compound I on .ompuId. 5 vels i SNS-59 Entry SoA C Compound I Compound 5 0,75 k 0.60 0106 2 2A k 0k64 O7 3 5 k 0,24 0028 4 50g 0g3 0 0 9 5 50 g 02 0016 6 50g 0 0011 7 50g 0g 001I 8 1kg 0 001 Example 4~ Pharmacweutkcal Composition Sutable for ni ection or intravenouos Infusion and Determination of Partid Lnrity [0f1621 An iluIstrtive example of a suitable SNS 95 phamiaceutical composition acmprisys: 0 mug of substantially pre SNS~595 drug substance per ml- o aqueous sohuio of 4% sorbitol that is adjusted to pH 2.5 with methanesufionic acid. One protocol for making such. a solution includes the following for making a 100 rg/10 mi presentation: 100 nmg ubstantially pure SNSZ595 substance (prepred following fi methods described herein) and 450 mg o-sorbitol are added to distilled water; th.e volure is brought up to a volUme of 10 mln> and the pH of the resting solution is adjusted to 2.5 with mcthanesulfonic acid. [:001631 The resulting SNS-595 composition will be essentially free of visible parties and stable [001641 As noted, the presence of particulate matter can be detemned using ay converdent technique, Fon exanmple the presence of visikfe particles can be determined according to the method described by European Pharialcopia 5.0, Section 2 9.the entirety of wh ich is incorporated herein by reference Spueiaaly an apparatus having a vi wing station comprising (1) a. mart black panel of appropriate size held in a vertical -50position; (2) a rnon-glare white panel of appropriate size held in a vertical position next to the black panel; and (3) an adjustable lanmpholder cited with a suiZable, shaded, white1ight source and with a suitable light dittuser (esg. a viewg in mInator containing two 13 W fluorescent tubes, each 525 nn in length) is used, The intensity of illumination at the viewing point is maintained between 200) lux and 3750 lux. Adherent labels are removed from the container, The outside of the container is washed and dried on the outside. The container is gently swirled or inverted while ensuring that air bubbles are not introduced and the container is observed for about 5 seconds in front of the white panel to determine the presence of visibk particles. The container is then observed for about seconds in foat of the black panel to determine the presence of visible particles, If visible particles are detected; from viewing in front of either panel the corresponding container is rejected [00651 FMther the light obscuration particle count test described in USP~NI' Genera. Chuter 788 may be used, which is incorporated herein by reference in its entirety: Speifeally a suitable appratus based on the principle of light blockage which allows an automatic determination of the size of particles and the number of particles according to SIitgsd. The apparatus is calibrated using dispersions of spherical particles of known sizes between 10 pm and 2j pn USP Particle Count Reference Standard. thesee standard particles are dispersed in particle-free waters Care is taken to avoid aggzregzation of particles during dispersion, [001661 The test is carried out under conditions limiting particulate matter, for example in a laminar-ow cabinet The glassware and filtration equipment used is very careffIg washed, except for the membrane filters, with a warm detergent solution and rinsed with abundant amounts of water to remove al traces of detergent. Immediately before use. the equipment is rinsed from top to bottom, outside and then inside, with particle-free water t00 167} Care is taken not to introduce air bubbles into tbe preparation to be examined, especialy when fractions of the preparation are being transferred to the eontainr in which the determination is to be carried out. To check that the environment is suitable fIr the test. that the glassware is properly cleaned and that the water tO be used is particle-ree. th.e BlAing test is carried out: determine the particulate matter in S samples of particle fee water each. of5 -n-L. according to the method described below. If the lumber of particles of 10 pul or greater size exceeds 25 for the combined 25 mU the precautions taken for the test -51are not sufficient, The predatory steps are repeated until the environment.glassware and water are suitable for the test. 100168 The contents of the sample are mixed by slowly inverting the container 20 times scessivel If necessary the sealing closure is cautiously removed. The outer surfaces of the container opening are Weaned using a je. of particle-ee water and the closure is removed avoiding any contamination of the contents, Gas bubbles are eliminated by pRaliow tg tin. appropriate measures such as aloigtesample to stand for 2 in or by sonication, (001691 For small-volume parenteral products less than 25 mL in volume. the contents of 10 or more units are combined in a deaned container to obtain a volme of not less than 25 mL; the test solution may be prepared by mixing the contents of a suitable number of vials and dihting to 25 ml. with particlefree water or with appropriate partice4free solvent when particlikee water is not suitable. Parc.nteral products having a vohune of 25 ml or more may be tested individually. The number of test specimens is such to afford atatistically sound a.ssessme;.For large~ volume parenterais or for sma -volume parenterals having a volume of 2$ mL or more. fewer than i 0 units may be testedbased on an appropriate sampling pan. 1001701 Four samples are removed from the specimen. each ofnot less'than 5 m." iand the particles having diameters equal. to or greater than each of 1.0 pim and 25 pa are counted. The result obtained for the first sample is disregarded and the mean number of parties for the specimen is calculated Examle :Synthesis of Comnound 4 H0 .. $N C - O 2 Et C 7HO C N N c0 N N 14N Sl S 4 [001 7.lj Compound - (00 g; 1,0 eq) was added to a solution of 25 g of' hiOH-HN) (0.2 5 eq) in 62$ mL. water and 125 mL EtOH. The resulting slurry was mixed was stirred at 25 30 "C overnight. The reaction mixture was then filtered and washed with 500 ml- water and 500 ml EtOH T he solids were then dried at 50 C overnight in acuto todrd o.poun.d 4, 'H NIR (400 MHz DISO-dj): 6 95 (s I H) 8.5 (d = 8.4 Hz. I H'h 7.95 (d. = 4 52- Hz, H), 7,88 (d,J= 32 Hz IDH) 7,46(dJ=8 88 HzH 1) 'C NMR (125 MHNz DSO d) 193 0, 1712, 1.62.0, 155.6. 153,7, .152,0, 140,9 137,99 1.24.4, 1.19,9, .110,9, 106,4; MS m/z 310 308 mdl) Lsamnple 6: Sx'nthesis of Comp ound 6 OH 'C, OH OH H -N O HO ,' N N SN 4 6 [00121 Comunpound 4 (0 g) was taken in 500 mL acetic acid rtesuling slurry was stirred for 15 minutes (uin) at RT under nitrogen atmosphere The slurry was then added to a 10 L. 3-ecked round bottom flask containing 4,5 L of 37% aqueous formaidrhyde solution. The resung sirry was then stirred at 6062 C overnight and. cooled to 25-30 'CThe reaction mxturc was then filtered, and the resulting white powder was dried under vacuum t 25-30 C overnight The solid was raesuspended in 150 m acet acid and tired for 3 days at RT. TIe resulting slury was then filered, and the solids were dried in a vacuum overight to afford Compound 6. H NR (400 MHz DMSO-d ) 8.38 (d= 8.8Hz, 2H), 797 (d.,J= 36 Hz, 2H1%7,89 (d, </= 36 Hz, 2H ), 7.6 (dU J8 Hz, 211), 3.89 (A 2); MS: n'z 571, 573, 575 (M+H)0. (001731 Compound 6 was analyze ed by polarized light microscopy to assess its crystalliniy and crystalline habit. Compound 6 appears to be a crysta lline powder under polarized light microsCope as it exhibits strong birefrjigence 1he crystaliine habit is that of pates 50400 pn. as isolated fro. acetic acid Enple 7: Preparation of Substandally Pure SNS-595 Substance from 26 Dichkoronicntinic Acid and N-Boc-3-'rroline [001741 Pgpaationofonpo'indifromZhicloroniggdniagi 100175j A solution of carbonyvdimmdazoie(CD l(64 kg) in tetrahydrofarani THF added to a slurry of 2.,6-dichloroicotnie acid (Conmpound A) (i? kg) in THE After about 21h, ethyl potassium in naonate (Elf ((02 CCC0K) (19.4 kg), triethylamrine (2.5.9 kg) and magnesium chloride (11.9 kg) were added and the reaction stirred for about 24 I. The reaction mixture was quenched wIth dihte FTC and extracted with ethyl acetate. The organic layer was concentratedashed with a mixture of aqueous NaG and NaHCO3. The organic aver was diluted with methylcyclonexane and dred by yacuum dtlalion, The sohition was treated with triethylrthoformate (17J kg)and acetic anhydride (59 kg) at about 90 to 10 *C, After the reaction was judged to be complete the excess ace tic anhydride was removed by distillations wth methyl.cyciohexane. TIe crude product was treated with a solution of 2eaminothiazole (v.2 kg) in T HF After about 2 hr. the reaction mixture was treated with potassium carbonate 03.6 kg) and the mixture stirred fr about 6 hr. The product wa prcpittd by te.addi isolated by filtration, washed iv~ , jth .d to ('nwun c CoI vit ACN-water, ACN and dried to giveC 1 1 kg) 0 NC N N CI N C1 N 1001761 Jr n, of aomond& NBoc -Yri 1001771 (ioU00romo-4hydrox)pyroidine caboxylie aeid rerrbtyi ester (2), Tarakedbon Asynuny hi12 (2002)29892997) [00178 NAqocKpyrrohncR (296 g. 135 moes) was added to a slrry of]3-dibromo 5,5-dimethylhdantoin (270 g 0.94 moles) in acetonitrile (ACN. 1800niL) and water (296 mE, while maintaining the zerperature of the vessel at 0 to 10 'C, Afer the addition, the reaction mixture was warmed to RT and stirred umti the reaction was judged to be complete (TEC or HPI&}) The reaction was quenched by the addition of .5% aqueous sodium thiosulfate solion (600 ru.) and the product was extracted with diehloromethane (2x 750 mL) The combined organic layer was washed with water (300 mL) and brine (200 mL, The organic lay was dried over anhydrous Na 2 SOi (75 g) and concentrated under reduced pressure to give Compound 8 (450 g) which was directly used in the next step. 1001791 ariezacerhoxvlaneacid. e crgbu esterCompond D An aqueous solution of sodium hydroxide (NaGH 155 IL, 2N) was added to Compound C (450 g, L69 moles) and the reaction was sired between for 2 hr at about RtT. The product was extracted with diebloromethane (2 x 1,25 L) and the conthined orgaie -54iayer was washed with waten (2 750 L) to neutral pH and then dried over anhydrous NaaSO. Evaporation under reduced pressure gave the epoxide D(9L0 go 1001801 (4~41vdox 4.gthylmilggymrlidine-1_-earggyfcid k ~1Jestar (Coame Aqueou s methyulaine solution (40% solution i ' mL, 38 mol) was added to the epoxide 1) (140 g, 0 65 unol) at RT and the reaction Was stirred until complete The excess rethylamine was removed by distilation;mder reduced pressure. To the residue obtained, diisopropyl ether (800 mL) was added and the mixture stirred fbr about 30 mBY The solid was filtered, washed with diisopropyl ether (200 l) then dried to give Compound E (135 g). [00181] ()Hvdroxy4-methylaminopyrroli dine- c-arboxy lie acid, tert-butyi ester (Compound E)X fronm Compound Ci [0182j Ten grams (10 g}of bromohydrin (Compound C) was treated with 40% aqueous netylamine (30 Ml>) and sodium bicarbonate (3.1g) at RT to givCompound E (8.g)4 butvl ester. usint -malie acid. TIhe aminoalcobol (Compound EA (100 ,a 0.46 moles) was dissolved in a mixture of aceton (600 m.) and water (1 mi ) at RiT The reaction mixture was heated to about 40 C and L+(->maie acid (62 g 0.48 moles) was added, The mixture was heated to about 50 to 55 *C to form a clear solution and then gradually cooled to RI' and then to 5 to 10 *C. The crystals formed were filtered, washed with acetone (2 a 70 mL), and dried under reduced pressure to give the imalate salt F (60 g, 37%)with purity by chiral EIPLC ratio ot S to R enantiomers (S;R 100:0, [001841 A small sample was analyzed for enantiomerie purity by conversion to Compound C and analyzing the resulting Compound G by chiral OP(Chiracel OD-H SC(522; mobile phase; hexane:IPA 95.5; I mL/min. The retention time for the 5 enantiomer is 7.7.25 min. 1001 85J Rtesolution of ft 3-Hyl droxv-4-metvlamnino-pvrroi dine- I carboxviec acid te uMyterusing()~Wyvrgglutanc d Reooution of Compound E (10 g) wi (L) ) pyroghutani acid (158 g in acetone (120 ml) and water (4 mi.) gave the pyroglutamat' salt (5.7 g) Crystallization from acetone-water gave 4.2 g of the PGA salt with 946 raio of diastereomers .An additional recrystalization from acetone-water gave the diastereomerically pure PGA salt (23 g, > 99% de). -55-.

[001861 Preparation of 3~(terFButoxycarbonyl-methy-anino)- roxy-pyrrohdine& carboxylic acid crbutyl ester (Compound G) from ( -)-malic acid salt (Compound F). [001871 To a rtaure of Compound F (57 g, 0 16 mois) in. methanol (MoOSl 220 mI) KC038(68 OA9t moles) was added at RT'. Boc anhydride (40 g, 0,18 moles) was added dropwise to the reacdon mixture over about 1 hr and the reaction mixture was strred until the reaction was complete (about 2 r1 Methanol was distilled off under reduced pressure at about 55 to 60 C, water (150 nmL.) was added to the reaction mixture and. the product was extracted with methyl erabutyl ether (MTBE 2 x 150 mid The combined organic layer was washed with water (200 ml) and brine 100 n L and then dried over anhydrous NaSO Concntration under reduced pressure gave Compound G as a white solid (52 g), [O1881 haryycaronlo-ymethy amno4q-nelhoxv-~pyroliinecrboxvlic acidjca ltxIewi n uiundHj 10441W S .A suspnsion of Conpound G (52 g, 0 16 ol) in THF (150 ml) was stirred at R for about 30 nin and cooled to 10 to ~15 0 C A solution of potassi mr hexamethyldisily amide (KHMDS. 40 solution in TFE, 144 mL, 0.256 mol) was slowly added while operature between-5 and 15 $ A fer 15 iidimnethyl sulfate (187 t, 1.20 mo l)was added dropwise to the reaction mixture while maintaining temperature between -10 and 0 'C and. the resulting reaction mixture was then stirred at this temperature for about 30 mi, The reaction mixture was quenched by the addition of water (1 00 ml.1 lowed by acetic acid (50 mn.), The product was extracted with methyl tert butyl ether (x 150 nml ) The combined organic layer was washed with water (100 nEl brine (50 ml) an dried over anhydrous NaSO 4 . Evaporation mder reduced pressure gave Compound H as an oil (54 g). [00190} )+ 4-Mthoxvaprroidin3vD-methl.amine (Cotmpound 2). renared using fal suld aid ( 1. To a solution of Compnound.U (540 , 0.163 mnoles) in THF (180 ml>) and MeOH (90 mi).p-toluenc sulfonic acid nmonohydrate (84 g, 0142 moles) was added and the reaction mixture was heated to 55460 C for about 5 hr, at which time the deprotection was complete. After cooling to about 4 0( 45

G(

1 , 0.2 g seed crystals of Compound 2 was added to the reaction mixture resulting in immediate rystalization. The slurry was ma-intained at 40-45 Chfr about 30 minutes and then gradualy cooled to 0 -5 *C, After agitating for 2 hr at 05C solids were filtered, washed with THF (2 x 50 omt), -56and dried to give the tosylate saIt Campound 2 as a white solid (66 g) with HPLC purity= 98.9%. [0191} The HPLC conditions were as followsColumn: Chiraiee& AD - H SC\523; nie phase: leptan: IPA (05%TPA) 85:15; flow rate: 1.0 mrLnm and runtime: 20 mnn f0 I92j Comtpound 2 has the rfention twe of I2. rin. Enantiomneri excess of iis material was greater than. 99%cee - .?. ........ -C E [001931 Prprto ofsbtniMueLN-9 usaceyarpoesn [00 194 1 To a slurry of Compounad 2 (8A kg) iN ACN at about 5 D PIEA N.8kg) is adde-d, After about 15 mtin, Campouind I (SA) kg) is added to the -reaction i:ixture, TheI recion mi ,-xtureis heated to ,bout 45 C for about 3 rs coole an theprdu tmrd The filtr cake is wsewin AC and deeJtd to gi1e ComOUnd 3, [00195 Tro a sohition of NaOH (0 8 kg! in water N1 5, k); Comapound 3 (5,5 kg) anid E-1OH (0.5kg) are, added The reaction mixture isfiltered and the fikateacdified to pH- 7-3-73 bay the addition of acetc acid. The mixture is then, heated to about, -,,5 -65 *C for6i about 2 br, After- cooting to amnbiewn mhure, the re ,actkinetr i, fdered. and wwshed wihvate~r and then with EtOEL T['he ikr ake is drevudracuum. T'he en-tde product is slu-tried in EtOHI at Aou 80C A. flrcogbFpouti (Urd vase with EDOH and dried to give a SNS4-"95 mixture 1001961 Nqext, to a solution of NaOH (0,2 k)in water (4 kg), the S NS-595 mnixturex obtained above (1 49 kg) and ERH (M 1.3 kg) a e, aded,Thmatonitresfhrd -57~ and the filtrate acidified to pH 73-7 by the addition of aqueous acetic acid (prepared from O9kg acetic acid and 2 9kg water) The mixture is then heated to about 55-65 'C for about 2 hr, After cooing to ambient tmperatie, the reaction mixture is filtered and washed with water and then with EtOH The filter cake is dried under vacuum. The crude product is scurried in EtOH at about 80 ! Ater coolI, the product is filtered, washed with EtOHI and died to give a substantiallypure SNS-55 substance [19 71 repair of suanalipure S -ancevia wv diisopylth I amine. [00198 To a surry ofCompound 2 (.55 kg) in acetonitrile (ACN; 10 L) diisopropylethylanine DIPEA; 4 ) and water (O.5 ) were added. To the solution Compound 1 ( kg) and acetonitril (I Q were added and the reaction was stirred for about 12 hr at RT The reaction mixture was then heated to about 45 TC, for about 2-6 hr After cooling, the product was filtered, washed with ACN (4 L and dried under vacuiun to give Compound 3 (1 kg) 1PLC analysis showed isimaterial to contain 01% Compound 1. [001991 To a solution of Na H (0135 kg) in water (3 3 I.substantially pure Compound 3 (09 kg) and EtCH are added. Aner hydrolysis was complete, the reaction mixture was filtered and the filtrate acidified to pH 73-7 by the addition of aqueous acetic acid, The mixture was then heated to about 55~65 *C7 tor about 2 hr. After cooln to ambient temperature the reaction mixture was filtered and washed with waer and then with EtOH The filer cake is dried under vacuunm The crude product was slurried in EtOH at about 80 After cooling, the product was filtered, washed with EtIHCl and cried to give a substantially pure SNS -595 substance (t 66 kg) [002001 The embodiments ofthe claimed subject matter described above are intended to be merely exemplary, and those skilled in the art will recognize or will be able to ascertain using no more than routine experimentation.nmerous equivalents of specific compounds, materials, and poeedures. All such equivalents are considered to be within the scope of the claimed suliec matter and are encompassed by the appended claims. -58- [00201] In the claims which follow and in the preceding description of the invention, except where the context requires otherwise due to express language or necessary implication, the word "comprise" or variations such as "comprises" or "comprising" is used in an inclusive sense, i.e. to specify the presence of the stated feature but not to preclude the presence or addition of further features in various embodiments of the invention. [00202] It is to be understood that, if any prior art publication is referred to herein, such reference does not constitute an admission that the publication forms a part of the common general knowledge in the art, in Australia or any other country. - 59 -

Claims (15)

1. A composition consisting of a substantially pure (+)-1,4-dihydro-7 [(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8 naphthyridine-3-carboxylic acid substance, wherein the substantially pure (+)-1,4 dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2 thiazolyl)-1,8-naphthyridine-3-carboxylic acid substance comprises (+)-1,4-dihydro

7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8 naphthyridine-3-carboxylic acid, or a salt or solvate thereof, and 0 to 0.02% Compound 4 OH 0 cl H N N 4 and less than 1.0% of other compounds based on total weight of the substantially pure (+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrolidinyl]-4-oxo-1-(2 thiazolyl)-1,8-naphthyridine-3-carboxylic acid substance. 2. The composition of claim 1, wherein the substantially pure (+)-1,4 dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-l-pyrrolidinyl]-4-oxo-1-(2 thiazolyl)-1,8-naphthyridine-3-carboxylic acid substance further comprises 0 to 0.02% Compound 5 0 0 GI XKYoH N N 5 based on total weight of the substantially pure (+)-1,4-dihydro-7-[(3S,4S)-3-methoxy 4-(methylamino)-1-pyrrolidinyl]-4-oxo-l-(2-thiazolyl)-1,8-naphthyridine-3 carboxylic acid substance. 3. The composition of claim 2, wherein the substantially pure (+)-1,4 dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2 thiazolyl)-1,8-naphthyridine-3-carboxylic acid substance comprises 0 to 0.03% of - 60 - Compound 4 and Compound 5 combined based on total weight of the substantially pure (+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo 1-(2-thiazolyl)-I,8-naphthyddine-3-carboxylic acid substance. 4. The composition of claim 1, wherein the substantially pure (+)-1,4 dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2 thiazolyl)-1,8-naphthyridine-3-carboxylic acid substance comprises 0 to 0.01% Compound 4 based on total weight of the substantially pure (+)-1,4-dihydro-7 [(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8 naphthyridine-3-carboxylic acid substance 5. An aqueous solution comprising the composition of claim 1. 6. The aqueous solution of claim 5, comprising methanesulfonic acid in an amount to provide the solution a pH of 2.5 7. The composition of claim 1, wherein the substantially pure (+)-1,4 dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2 thiazolyl)-1,8-naphthyridine-3-carboxylic acid substance further comprises 0 to 0.1% Compound 7 HOe 0 2 H NN 7 based on total weight of the substantially pure (+)-1,4-dihydro-7-[(3S,4S)-3-methoxy 4-(methylamino)-1-pyrrolidinyl]-4-oxo-l-(2-thiazolyl)-1,8-naphthyridine-3 carboxylic acid substance -61 -

8. An aqueous solution comprising the composition of claim 1, wherein the substantially pure (+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1 pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylic acid substance further comprises 0 to 0.02% Compound 6 OH OH CIl NN 0 N N CI SAN N'S 6 based on total weight of the substantially pure (+)-1,4-dihydro-7-[(3S,4S)-3-methoxy 4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3 carboxylic acid substance.

9. The aqueous solution of claim 8, wherein the substantially pure (+) 1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2 thiazolyl)-1,8-naphthyridine-3-carboxylic acid substance comprises 0 to 0.01% Compound 6 based on total weight of the substantially pure (+)-1,4-dihydro-7 [(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazoly)-1,8 naphthyridine-3-carboxylic acid substance.

10. The aqueous solution of claim 8, wherein the aqueous solution is essentially free of sub-visible and visible particles for at least one month starting from the time of production.

11. The composition of claim 1, wherein (+)-1,4-dihydro-7-[(3S,4S)-3 methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-I.-(2-thiazolyl)-l,8-naphthyridine-3 carboxylic acid is present as a hydrate.

12. An aqueous solution comprising the composition of claim 1, wherein about 100 mg of substantially pure (+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4 (methylamino)-1-pyrrolidinyl]-4-oxo-l-(2-thiazolyl)-1,8-naphthyridine-3-carboxylic acid substance, or a salt or solvate thereof, is present for every 10 mL of the composition, and wherein the aqueous solution is stable at 3 months after production.

13. The aqueous solution of claim 12, wherein the aqueous solution has not more than 1000 particles > 10 microns per 10 mL of the aqueous solution. - 62 -

14. An aqueous solution comprising the composition of claim 1, wherein the aqueous solution consists of 100 mg substantially pure (+)-1,4-dihydro-7 [(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8 naphthyridine-3-carboxylic acid substance, or a salt or solvate thereof, 450 mg sorbitol, water, and methanesulfonic acid, wherein the methanesulfonic acid is present in an amount to adjust the pH of the aqueous solution to about 2.5, and wherein the water is present in an amount to provide a total solution volume of 10 mL.

15. The composition of claim 1 wherein the substantially pure (+)-1,4 dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2 thiazolyl)-1,8-naphthyidine-3-carboxylic acid comprises 0 to 0.02 wt% Compound 4 OH 0 CI N H NN CI N N 0 4 and 0 to 0.1 wt% Compound 7 H02C 02M ~ H -0 N N C HS 7 based on total weight of the composition.

16. The composition of claim 1 wherein the substantially pure (+)-1,4 dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2 thiazolyl)-1,8-naphthyridine-3-carboxylic acid comprises 0 to 0.02 wt% Compound 4 OH 0 cl. H CIN N 0 SN 4, - 63 - 0 to 0.02 wt% Compound 5 0 0 cl YOH CIN N AN 5, and 0 to 0.1 wt% Compound 7 HO CO 2 H CH~ 7 based on total weight of the composition

17. An aqueous solution comprising the composition of claim 1, wherein the substantially pure (+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1 pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylic acid substance consists of (+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4 oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylic acid, or a salt or solvate thereof, 0 to 0.02 wt/o Compound 4 OH 0 ~ N H CI N 0 S N 4, o to 0.02 wt% Compound 5 0 0 OH N N 5, - 64 - and 0 to 0.1 wt% Compound 7 11O, Ci 02H1 H2 N N cH 7, based on total weight of the substantially pure (+)-1,4-dihydro-7-[(3S,4S)-3-methoxy 4-(methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3 carboxylic acid substance.

18. A substantially pure (+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4 (methylamino)-1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylic acid substance consisting of (+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino) 1-pyrrolidinyl]-4-oxo-1-(2-thiazolyl)-1,8-naphthyridine-3-carboxylic acid, or a salt or solvate thereof, and 0 to 0.02% Compound 4 OH 0 cl- H CIN N 0 .AN 4 and less than 1.0% of other compounds, based on total weight of the substantially pure (+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1-pyrrolidinyl]-4-oxo 1-(2-thiazolyl)-I,8-naphthyridine-3- carboxylic acid substance.

19. A composition according to any one of claims 1-4, 7, 11, 15 and 16, or a substantially pure (+)-1,4-dihydro-7-[(3S,4S)-3-methoxy-4-(methylamino)-1 pyrrolidinyl]-4-oxo-l-(2-thiazolyl)-1,8-naphthyridine-3-carboxylic acid substance according to claim 18, substantially as herein described or exemplified.

20. An aqueous solution according to any one of claims 5, 6, 8-10, 12-14 and 17, substantially as herein described or exemplified. - 65 -