MXPA98003316A - Procedure for preparing a secretagogo de hormona de crecimie - Google Patents

Abstract

The present invention is directed to a novel process for the preparation of the compound N- [1 (R) - [(1,2-dihydro-1-methanesulfonyl-spiro- [3H-i ndol-3,4-pipe-ridin] - 1'il) carbonyl] -2-phenylmethyl- or xy) ethyl] -2-amino-2-methyl-propanamide, and salts thereof, having structure (1) and having the ability to stimulate hormone release of natural or endogenous growth, this compound can be used to treat conditions that require the stimulation of production or secretion of growth hormone such as in humans with a deficiency of natural growth hormone or in animals it can be used for food production or wool where stimulation of growth hormone results in a larger and more productive animal

Description

PROCEDURE FOR PREPARING A SECRETAGOGUE OF GROWTH HORMONE BACKGROUND OF THE INVENTION Growth hormone, which is secreted from the pituitary, stimulates the growth of all the tissues in the body that are able to grow. In addition, it is known that growth hormone has the following basic effects on the metabolic processes of the body: (1) increased speed of protein synthesis in all cells of the body; (2) decreased speed of carbohydrate utilization in body cells; (3) increased mobilization of free fatty acids and use of fatty acids to obtain energy. A deficiency in the secretion of growth hormone can result in several medical disorders, such as dwarfism. Several pathways that release growth hormone are known. For example, chemical compounds such as arginine, L-3,4-dihydroxyphenylalanine (L-DOPA), glucagon, vasopressin and insulin induced by hypoglycemia, as well as activities such as sleep and exercise, indirectly cause the hormone of growth is released from the pituitary acting in some way on the hypothalamus, perhaps to decrease the secretion of somatostatin or to increase the secretion of the known secretagogue releasing factor of growth hormone (GRF) or an unknown hormone releasing hormone endogenous growth, or all of them. In cases where increased levels of growth hormone were desired, the problem was usually solved by providing exogenous growth hormone or by administering GRF or a peptide compound that stimulated the production and / or release of growth hormone. In any case, the peptide nature of the compound required that it be administered by injection. Initially, the source of growth hormone was the removal of the pituitary glands from corpses. This resulted in a very expensive product and brought with it the risk that a disease associated with the source of the pituitary gland could be transmitted to the growth hormone receptor. The recombinant growth hormone has now been obtained and, although it does not entail any further risk of disease transmission, it is still a very expensive product that must be administered by injection or by nasal spray. Other compounds that stimulate the release of endogenous growth hormone have been developed. In particular, certain spiro compounds are described in PCT patent publication WO 94/13696 and Proc. Natl.

Acad. Sci. USA. 92, 7001-7005 (July 1995) as non-pethidic secretagogues of growth hormone. These compounds have the ability to stimulate the release of natural or endogenous growth hormone, and can thus be used to treat conditions that require stimulation of the production or secretion of growth hormone, such as in humans with a growth hormone deficiency. natural, or in animals used for the production of food or wool, where the stimulation of growth hormone will result in a larger and more productive animal. Among the preferred compounds described in the present invention, there is spiro [3 H -indole-3,4'-piperidin] -l '-yl) carbonyl] -2- (phenylmethyloxy) ethyl] -2-amino-2-methylpropanamide, which has the structure: PCT patent publication WO 94/13696 describes methods for preparing this compound (see Examples 18, 19 and 55). However, the synthesis of the compound was achieved using the very expensive amino acid coupling agent EDC ($ 1100 / kg); the use of numerous equivalents of trifluoroacetic acid as a catalyst for deprotections of the BOC group; and extensive chromatographic purifications; and resulted in "thickening" of the final product. The advantages of the present invention include: a 6-step high yield non-insulating process that provides > 99.9% purity; reduced cost due to the use of DCC [$ 40 / kg] instead of EDC [$ 1100 / kg]; environmental impact reduced by the use of methanesulfonic acid in place of trifluoroacetic acid as a catalyst (as well as minor equivalent of catalyst) in the deprotections; and ease of isolation of the final product.

BRIEF DESCRIPTION OF THE INVENTION The present invention is directed to a process for preparing the compound N- [1 (R) - [(1,2-dihydro-l-methanesulfonyl-pyrro [3 H -indol-3,4'-piperidin] -l'-yl) carboni l] -2- (f-enylmethyloxy) ethyl] -2-amino-2-methyl-propanamide, which has the structure: and salts thereof, in particular, the methanesulfonate salt. This compound has the ability to stimulate the release of natural or endogenous growth hormone, and can thus be used to treat conditions that require the stimulation of the production or secretion of growth hormone, such as in humans with a growth hormone deficiency. natural, or in animals used for the production of food or wool, where the stimulation of growth hormone will result in a larger and more productive animal.

DESCRIPTION OF THE INVENTION The present invention is directed to a novel process for preparing the compound N- [1 (R) - [(1,2-dihydro-l-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -l'- il) carbonyl] -2- (phenylethyloxy) ethyl] -2-amino-2-methyl-propanamide, which has the structure: and salts thereof, in particular, the methanesulfonate salt. The present process provides the desired compound from inexpensive, readily available and environmentally acceptable starting materials, reagents and solvents. The procedure does not require the use of any chromatographic purification, and it is possible to obtain the final product from the intermediate spiroindoline sulfonamide without isolation of any of the intermediates. The individual procedures within the general procedure are summarized as follows: SCHEME I and SCHEME I (CONTINUED) Within this general procedure, a first method relates to the preparation of a compound of the formula I: wherein L is a protecting group of amino group, by coupling an amino acid of the formula: Ph '. ^ N / CO2H N-L H with a compound of the formula: in the presence of an acid activating agent in an inert solvent in the presence of a catalytic agent, to give the compound of formula I. Acid activating agents suitable for this process include: DCC, EDC, ECAC and BOP, wherein the preferred acid activating agent is DCC (N, N'-dicyclohexylcarbodiimide). Catalytic agents suitable for this process include: HOBT and the like, in which a preferred catalytic agent is HOBT (hydroxybenzotriazole). Suitable inert solvents for these processes include: acetonitrile; isopropyl acetate; ethyl acetate; propionitrile; Water; chlorinated hydrocarbons such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene, ortho-dichlorobenzene; benzene; toluene; xylenes; and the like; and mixtures thereof, wherein the preferred solvent is acetonitrile or isopropyl acetate and water.

The preferred scale of the reaction temperature is between -40 and 150 ° C, and the most preferred scale is between 20 and 35 ° C. Suitable protecting groups of amino group include: benzyl, benzyloxymethyl, benzyloxycarbonyl (carbobenzyloxy), benzylsulfonyl, 2-bromo-ethyloxycarbonyl, t-butoxycarbonyl, 2-chloro-benzyloxycarbonyl, 2-chloroethyloxycarbonyl, di-t-amyloxycarbonyl, 9-f luoroenyl-methyloxycarbonyl, isopropoxy carbonyl, 4-methoxy-benzyloxycarbonyl, -nitrobenzyloxycarbonyl, 2-nitrophenyl-sulfonyl, phthaloyl, 2,2,2-trichloro-t-butyloxycarbonyl, trifluoroacetyl, triphenyl ethane, allyloxycarbonyl and vinyloxycarbonyl, and the like, wherein the most preferred include benzyloxycarbonyl (carbobenzyloxy) and t- groups butoxycarbonyl, and in which the most preferred is the t-butoxy carbonyl lo group. As far as efficiency is concerned, it is preferred that this coupling be carried out in situ without isolating the compound of formula I, after its preparation by the aforementioned process. Within this general procedure, a second process relates to the preparation of a compound of formula II: which comprises reacting a compound of the formula I: I wherein L is an amino group protecting group, with an amino group deprotecting agent to give the compound of the formula II. Suitable protecting groups of amino group include: benzyl, benzyloxy ethyl, benzyloxycarbonyl (carbobenzyloxy), benzylsulfonyl, 2-bromo-ethyloxycarbonyl, t-butoxycarbonyl, 2-chloro-benzyloxycarbonyl, 2-chloroethyloxycarbonyl, di-t-amyloxycarbonyl, 9-fluoroenil -methyloxycarbonyl, isopropoxycarbonyl, 4-methoxy-benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 2-nitrophenylsulfonyl, phthaloyl, 2,2,2-trichloro-t-butyloxycarbonyl, trifluoroacetyl, triphenylmethane, allyloxycarbonyl and vinyloxycarbonyl, and the like, in the that the most preferred include benzyloxycarbonyl (carbobenzyloxy) and t-butoxycarbonyl groups, and in which the t-butoxycarbonyl group is most preferred. In this process, the removal of the amino group protecting group can be achieved by the use of an appropriate catalytic agent. The removal of a t-butoxycarbonyl protecting group can be carried out in a solvent such as methanol, ethanol, methylene chloride, ethyl acetate or isopropyl acetate, with a strong acid. Such strong acids include methanesulfonic acid, trifluoroacetic acid, hydrochloric acid, hydrogen chloride gas, hydrogen bromide; hydrogen iodide; trifluoro-ethanesulfonic acid; camphor sulfonic acid; sulfuric acid; phosphoric acid; and an arylsulfonic acid such as benzenesulfonic acid, p-toluenesulfonic acid and p-chlorobenzenesulfonic acid. Preferred catalytic agents include: trifluoroacetic acid; methanesulfonic acid; camphor sulfonic acid; benzenesulfonic acid; p-toluenesulfonic acid; and p-chlorobenzenesulfonic acid. The most preferred catalytic agent is methanesulfonic acid. The preferred solvent is methanol or ethanol, and the most preferred solvent is ethanol. The preferred scale of the reaction temperature is between -40 and 150 ° C, and the most preferred scale is between 10 and 40 ° C. The removal of a benzyloxycarbonyl group (carbobenzyloxy) can be achieved by various methods, for example, catalytic hydrogenation with hydrogen in the presence of a noble metal or its oxide, such as palladium on activated carbon in a protic solvent such as ethanol. In cases where the catalytic hydrogenation is contraindicated by the presence of another potentially reactive functionality, the removal of the benzyloxycarbonyl group (carbobenzyloxy) can also be achieved by treatment with a solution of hydrogen bromide in acetic acid, or by treatment with a mixture of TFA and dimethyl sulfide. As far as efficiency is concerned, it is preferred that this acid-catalysed deprotection be carried out in situ without isolating the compound of the formula II after its preparation by the aforementioned process. Within this general procedure, a third method relates to the preparation of a compound of formula III: wherein L is a protecting group of amino group, by coupling an amino acid of the formula: wherein L is an amino group protecting group, with a compound of formula II: II in the presence of an acid activating agent in an inert solvent in the presence of a catalytic agent, to give the compound of formula III. Acid activating agents suitable for this process include: DCC, EDC, ECAC and BOP, wherein the preferred acid activating agent is DCC (N, N'-dicyclohexylcarbodiimide). Catalytic agents suitable for this process include: HOBT and the like, in which a preferred catalytic agent is HOBT (hydroxybenzotriazole). Suitable inert solvents for these processes include: acetonitrile; isopropyl acetate; ethyl acetate; propionitrile; Water; chlorinated hydrocarbons such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene, ortho-dichlorobenzene; benzene; toluene; xylenes; and the like; and mixtures thereof, wherein the preferred solvent is a mixture of isopropyl acetate and water, preferably in a ratio of about 40:60 to 60:40 (by volume), and more preferably in a ratio of about 50: 50 (in volume). The preferred scale of the reaction temperature is between -40 and 150 ° C, and the most preferred scale is between 20 and 50 ° C. Suitable protecting groups of amino group include: benzyl, benzyloxymethyl, benzyloxycarbonyl (carbobenzyloxy), benzylsulphonyl, 2-bromo-ethyloxycarbonyl, t-butoxycarbonyl, 2-chloro-benzyloxycarbonyl, 2-chloroethyloxycarbonyl, di-t-amyloxycarbonyl, 9-fluoro- ethyloxycarbonyl, isopropoxycarbonyl, 4-methoxy-benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 2-nitrophenylsulphonyl, phthaloyl, 2,2,2-trichloro-t-butyloxycarbonyl, trifluoroacetyl, tri-phenylmethane, allyloxycarbonyl and vinyloxycarbonyl, and the like, in which most preferred include benzyloxycarbonyl (c-rbobenzyloxy) and t-butoxycarbonyl groups, and in which the t-butoxycarbonyl group is most preferred. As far as efficiency is concerned, it is preferred that this coupling be carried out in situ without isolating the compound of formula III, after its preparation by the aforementioned process. Alternatively, the compound of formula III can be isolated as a defined intermediate. Within this general procedure, a fourth method relates to the preparation of a compound of formula IV, or a pharmaceutically acceptable salt thereof: IV comprising reacting a compound of formula III wherein L is an amino group protecting group, with an amino group deprotecting agent to give the compound of the formula IV. Suitable protecting groups of amino group include: benzyl, benzyloxymethyl, benzyloxycarbonyl (carbobenzyloxy), benzylsulfonyl, 2-bromo-ethyloxycarbonyl, t-butoxycarbonyl, 2-chloro-benzyloxycarbonyl, 2-chloroethyloxycarbonyl, di-t-amyloxycarbonyl, 9-f luoroenyl-ethyloxycarbonyl, isopropoxycarbonyl, 4-methoxy-benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl , 2-nitrophenyl-sulfonyl, phthaloyl, 2,2,2-trichloro-t-butyloxycarbonyl, trifluoroacetyl, triphenylmethane, allyloxycarbonyl and vinyloxycarbonyl, and the like, wherein the most preferred include benzyloxycarbonyl (carbobenzyloxy) and t-butoxycarbonyl groups, and wherein the t-butoxycarbonyl group is most preferred. In this process, the removal of the amino group protecting group can be achieved by the use of an appropriate catalytic agent. The removal of a t-butoxycarbonyl protecting group can be carried out in a solvent such as methanol, ethanol, methylene chloride, ethyl acetate or isopropyl acetate, with a strong acid. Such strong acids include methanesulfonic acid, trifluoroacetic acid, hydrochloric acid, hydrogen chloride gas, hydrogen bromide; hydrogen iodide; trifluoro-ethanesulfonic acid; camphor sulfonic acid; sulfuric acid; phosphoric acid; and an arylsulfonic acid such as benzenesulfonic acid, p-toluenesulfonic acid and p-chlorobenzenesulfonic acid. Preferred catalytic agents include: trifluoroacetic acid; methanesulfonic acid; camphor sulfonic acid; benzenesulfonic acid; p-toluenesulfonic acid; and p-chlorobenzenesulfonic acid. The most preferred catalytic agent is methanesulfonic acid. It is preferred that the compound of the formula V be isolated in the form of the methanesulfonate salt. The preferred solvent is methanol or ethanol, and the most preferred solvent is ethanol. The preferred scale of the reaction temperature is between -40 and 150 ° C, and the most preferred scale is between 10 and 40 ° C. The removal of a benzyloxycarbonyl group (carbobenzyloxy) can be achieved by various methods, for example, catalytic hydrogenation with hydrogen in the presence of a noble metal or its oxide, such as palladium on activated carbon in a protic solvent such as ethanol. In cases where the catalytic hydrogenation is contraindicated by the presence of another potentially reactive functionality, the removal of the benzyloxycarbonyl group (carbobenzyloxy) can also be achieved by treatment with a solution of hydrogen bromide in acetic acid, or by treatment with a mixture of TFA and dimethyl sulfide. As far as efficiency is concerned, it is preferred that this acid-catalysed deprotection be carried out in situ without isolating the compound of the formula II after its preparation by the aforementioned process. Within this general procedure, a fifth process relates to the preparation of a pharmaceutically acceptable salt of a compound of the formula IV, in particular, the methanesulfonate salt, that is, a compound of the formula V: which comprises reacting a compound of formula IV: with an acid, preferably methanesulfonic acid, to give the compound of formula V. It is preferred that the compound of formula V be isolated in the form of the methanesulfonate salt. The preferred solvent comprises ethyl acetate and ethanol, and the most preferred solvent is a mixture of ethyl acetate and ethanol.

As far as efficiency is concerned, it is preferred that the formation of the salt be carried out in situ without isolating the compound of the formula V after its preparation by the aforementioned process. In a preferred embodiment of the present invention, the individual procedures within the general procedure are described as follows: SCHEME II SCHEME II (CONTINUED) basic basic In this preferred embodiment, the CBZ-spiroindoline 1 is treated with Darco (20% by weight) before hydrogenation. The hydrogenation is carried out in ethanol at 65 ° C over 10% Pd / C with vigorous stirring. A solution of Ib in isopropyl acetate and water is coupled with commercially available N-BOC-O-benzyl-D-serine in the presence of dicyclohexylcarbodiimide (DCC) and 1-hydroxybenzotriazole (HOBt). After filtering the by-product of dicyclohexylurea (DCU), the phase 2 filtrate is separated, and the organic layer is washed successively with aqueous sodium hydroxide solution at MM, 0.5 M aqueous hydrochloric acid and finally with sodium hydrogen carbonate. watery saturated. Better results are achieved in this coupling when the solution of the free amino group in iPrOAc / H2? it is treated with DCC, HOBT followed by addition of the amino acid at room temperature, and followed by reaction for 3 to 5 hours. The batch is then concentrated in vacuo, and the solvent is changed from isopropyl acetate to ethanol. In general, this change of solvent proceeds quickly by "loading and extracting" three volumes per batch to remove the isopropyl acetate. The BOC group of 11 is removed by treatment with methanesulfonic acid (MsOH) (3eq) in ethanol at 35-40 ° C. The separation between isopropyl acetate and aqueous sodium hydroxide solution at ÍM produces 12. The coupling of 12 with N-BOC-α-aminoisobutyl acid is best performed in a two-phase solvent system, isopropyl acetate / water ( 1: 1) in the presence of DCC and HOBt (1.1 eq each). Removal of the DCU by filtration, separation of the layers, and washing the organic layer successively with aqueous sodium hydroxide to MM, 0.5 M aqueous hydrochloric acid and saturated aqueous sodium hydrogen carbonate, yields 14. The mixture is solvent changed by ethanol for the subsequent breakdown of methane sulfonic acid from the Boc group. The deprotection of 14 is more difficult than that of 11, and requires a concentrated solution of ethanol / methanesulfonic acid and heating at 35-40 ° C. After the extractable preparation (EtOAc-NaOH), free amine 15 is isolated.

The organic layer is washed well with 1 N NaOH to ensure complete removal of methanesulfonic acid. The ethyl acetate solution of the free base is concentrated to low volume in vacuo, and treated azeotropically on dry (KF <500 mgml-1) by "loading and extracting" two volumes per 90/10 batch of sodium acetate. ethyl / ethanol followed by two volumes per batch of ethyl acetate. The resulting slightly turbid dry solution of the free base in ethyl acetate is treated with Darco G-60 (25% by weight) at room temperature for about 10 hours. Removal of the Darco by filtration with a filtration agent gives the free base 15. The formation of the methanesulfonic acid salt 16 from 15 is carried out in EtOac with 1.1 eq of MsOH at about 50"C. Free 15 is treated with 8% by volume of EtOH and 1 eq of H2O, and heated to 55 ° C until complete dissolution.Cooling at room temperature and stirring the resulting suspension for 4 hours, gives a crystalline material of 16, designated as crystal shape II [solubility in IPA = 12 mg / ml]. The conversion of form II to form I is achieved where the salt is formed in EtOAc-EtOH as described above, but instead of cooling the initial solution of the salt (at 55 ° C) to room temperature, it is cooled to 45 ° C. The crystals should start to appear at that temperature, and the suspension should become denser over time. The temperature is then raised to 51 ° C, and the suspension is aged overnight. The complete conversion of Form I of 16 should be expected. Preferably, the conversion of Form II to Form I is accomplished by adding seed crystals of Form I to a solution of the free base in EtOAc-EtOH at 50-55pC followed by aging. Accordingly, the free base 15 can be treated with 1.1 equivalents of methanesulfonic acid in 8% ethanol in ethyl acetate at 50-55 ° C. The batch is then seeded with approximately 2% by weight of the form I of the methanesulfonate salt 16, and then aged at 55 ° C overnight. The batch is cooled to room temperature, and aged for about 2 to 3 hours. The product is isolated by filtration at room temperature under a nitrogen atmosphere, dried at 35 ° C in vacuo, and sieved to give the methanesulfonate salt 16. The methanesulfonic acid salt 16 can also be formed by alternating the gradual addition of MsOH (1.1 eq) and seed crystals of the form I to a solution of the free base in EtOAc-EtOH at about 50 ° C, where the order of addition of the MsOH and the seed is not critical. Throughout the present application, the following abbreviations are used with the following meanings: Bu Butyl Bn Benzyl BOC, Boc t-butyloxycarbonyl BOP Benzotriazol-1-yloxy tris (dimethylamino) phosphonium hexafluorophosphate Cale. Calculated CBZ, Cbz Benzyloxycarbonyl DCC N, N'-dicyclohexy Icarbodiimide DIEA Di-isopropylethylamine DMF N, N-dimethylformamide DMAP 4-dimethylaminopi ridine EDC 1- (3-Dimethylaminopropyl) -3- eti Icarbodiimide EDAC Ethyl-3- ( 3-dimethylamino) -propylcarbodiimide EI-MS Mass spectroscopy - electron ion Et Etyl Eq. Equivalent (s) FAB-MS Rapid atom bombardment - Mass spectroscopy h, hr. Hours HOBT, HOBt Hid roxybenzot riazole CLAP Chromatography of iPrOAc pressure fluids Isopropyl acetate KHMDS Bis (trimethylsilyl) potassium amide LAH Lithium-aluminum hydride LHMDS Lithium bis (trimethylsilyl) amide Methyl MF Molecular formula MHz Megahertz MPLC Chromatography of medium pressure liquids MsOH Methanesulfonic acid NMM N-methylmorpholine NMR Nuclear magnetic resonance Ph Phenyl Proline Prep. Prepared TFA Trifluoroacetic acid THF Tetrahydrofuran TLC Thin layer chromatography TMS Tetramethylsilane In the above structural formula, and throughout the present specification, the following terms have the indicated meanings: The phrase "peptide coupling reaction", as used in the present invention, means the coupling of a carboxylic acid with an amine using an acid activating agent such as EDC, DCC and BOP in an inert solvent in the presence of a catalyst such as HOBT. Suitable inert solvents for said couplings include: acetonitrile; isopropyl acetate; ethyl acetate; propionitrile; Water; chlorinated hydrocarbons such as dichloromethane, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene, orthodichlorobenzene; benzene; toluene; xylene; and combinations thereof; and similar. The variable "L" and the term "amino group protecting group" indicate the presence of a suitable protecting group of the amino group, such as those described in Greene, T.W., Wuts, P.G.M. Protective Groups in Organic Synthesis, 2a. ed., John Wiley & Sons, Inc. New York, 1991. An appropriate protective group will be able to withstand the reaction conditions of the intermediate procedures, before being removed, as desired. The amino group protecting group is independently selected for each procedure within the overall procedures. Suitable protecting groups of amino group include: benzyl, Benzyloxymethyl, benzyloxycarbonyl (carbobenzyloxy), benzylsulfonyl, 2-bromo-ethyloxycarbonyl, t-butoxycarbonyl, 2-chloro-benzyloxycarbonyl, 2-cloroetiloxicarbonilo, di-t-amyloxycarbonyl, 9-fluoroenil- ethyloxycarbonyl, isopropoxy carboni I, 4-methoxy- benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 2-nitrophenyl-sulfonyl, phthaloyl, 2,2,2-trichloro-t-butyloxycarbonyl, trifluoroacetyl, triphenylmethane, allyloxycarbonyl and vinyloxycarbonyl, and the like, in which more preferred include benzyloxycarbonyl (carbobenzyloxy) and t-butoxycarbonyl, and in which the most preferred is the t-butoxy carbonyl group. The removal of the amino group protecting group can be achieved by the use of an appropriate catalytic agent. The removal of a t-butoxycarbonyl protecting group can be carried out in a solvent such as methanol, ethanol, methylene chloride, ethyl acetate or isopropyl acetate, with a strong acid. Such strong acids include methanesulfonic acid, trifluoroacetic acid, hydrochloric acid, hydrogen chloride gas, hydrogen bromide; hydrogen iodide; trifluoromethanesulfonic acid; camphor sulfonic acid; sulfuric acid; phosphoric acid; and an arylsulfonic acid such as benzenesulfonic acid, p-toluenesulfonic acid and p-chlorobenzenesulfonic acid. Preferred catalytic agents include: trifluoroacetic acid; methanesulfonic acid; camphor sulfonic acid; benzenesulfonic acid; p-toluenesulfonic acid; and p-chlorobenzenesulfonic acid. The most preferred catalytic agent is methanesulfonic acid. The preferred solvent is methanol or ethanol. The removal of a benzyloxycarbonyl protecting group (carbobenzyloxy) can be achieved by various methods, for example, catalytic hydrogenation with hydrogen in the presence of a noble metal or its oxide, such as palladium on activated carbon in a protic solvent such as ethanol. In cases where the catalytic hydrogenation is contraindicated by the presence of another potentially reactive functionality, the removal of the benzyloxycarbonyl group (carbobenzyloxy) can also be achieved by treatment with a solution of hydrogen bromide in acetic acid, or by treatment with a mixture of TFA and dimethyl sulfide. The amine compounds used as starting materials for the process of the present invention may be present as their acid salts, such as the salts derived from the use of inorganic and organic acids. Examples of such acids are hydrochloric, nitric, sulfuric, phosphoric, formic, acetic, trifluoroacetic, propionic, maleic, succinic, malonic, methanesulfonic, and the like. Likewise, the compounds produced by the methods of the present invention can be isolated in the form of their pharmaceutically acceptable acid salts. In addition, certain compounds containing an acid function such as carboxy can be in the form of their inorganic salt, wherein the counterion can be selected from sodium, potassium, lithium, calcium, magnesium, and the like, as well as from organic bases. The preparation of compounds with the process of the present invention can be carried out in sequential or convergent synthetic ways. It is noted that, in some cases, the order to carry out the above reaction schemes can be varied to facilitate the reaction or to avoid inconvenient reaction products. In general, the process of the present invention is carried out sequentially as set forth in the present invention. Many of the starting materials are commercially available, or are known in the literature, and others can be prepared following methods described in the literature for analogous compounds. The skills that are required to carry out the reaction and purification of the resulting reaction products are known to those skilled in the art. Purification procedures include crystallization and reverse phase or normal phase chromatography. The following examples are provided solely for the purpose of improving the illustration, without pretending to be limiting of the disclosed invention.

EXAMPLE 1 N-benzyl carbamate of isonipecotic acid (3) Materials Isonipecotic acid (2) T.C.I. 4.02 kg (31.1 moles) Benzyl chloroformate (Schweitzerhall) 6.91 Kg (40.5 moles) K2CO3 10.1 kg (72.9 moles) Water 40.2 1 Isonipecotic acid (2) and K2CO3 were dissolved in 40.2 1 of water in a 4-neck flask of 100 1, with mechanical stirring under N2, and the solution was cooled to 10 * C. Benzyl chlorofomethyl was added, maintaining the temperature between 9 and 14 ° C, and the mixture was heated to 22 ° C after the addition was finished, and was aged for 58 h. The addition was concluded at 4 h, at which point the pH was 9.0. After aging for 58 h, there was no change in pH. The reaction mixture was transferred to a 200 1 extractor, and washed with 3 x 13 kg (15 1) of IPAC and 1 x 12 1 of EtOAc. The aqueous layer was extracted with 8 1 of toluene. After the washings, the benzyl alcohol content was reduced from 3.8% to 1.4% by CLAP analysis. Analytical CLAP: Zorbax Dupont 25 cm RXC8 column with 1.5 ml / min flow and detection at 254 nm; isocratic mixture with 35% MeCN, 65% aqueous H3PO4 at 0.1%; Retention times: 3 »= 6.9 min, benzyl alcohol = 3.3. min, toluene = 17.3 min. The aqueous phase was acidified with 37% aqueous HCl to pH 1.8. Carbon dioxide was produced during the addition of HCl, but gas production was easily controlled. The addition of HCl took < 1 h, and required 10 1 of concentrated HCl. The aqueous phase was extracted with 3 x 6.6 1 of toluene. The toluene extracts were dried with 2 kg of sodium sulfate, and filtered through a pad of Solka-floc ™. The combined filtrates weighed 17.8 kg. The crude yield of carbamate 3 was 7.89 kg (97%) (as obtained by evaporation of heavy aliquots of the filtrates to dryness). The filtrates were transferred through a lOμ in-line filter to a 100 1 flask. The extracts were concentrated at 10 mbar at < 25 ° C to a volume of 18 1. The final concentration of carbamate 3 was 440 g / 1. The concentration of the toluene filtrate served to azeotropically remove the final traces of water (final KF = 170 mg / l). The product was 99.1% pure in area with 0.9% in benzyl alcohol area as the only impurity.

EXAMPLE 2 N-benzyl carbamate of isonipecotic acid chloride (4) Materials: N-benzyl carbamate of isonipecotic acid (3) 7.89 kg (30.0 moles) in in toluene (P.M. = 263.30) 17.9 1 Oxalyl chloride (P.M. = 126.93) 3.94 kg (31.0 moles) DMF (P.M. = 73.10) 10 ml Toluene 12 1 To the toluene solution of benzyl carbamate 3 from the previous step, 5 ml of DMF and 10 1 of toluene were added. The oxalyl chloride was added over a period of 20 min. The reaction mixture was aged for 16 h at 18 ° C under a slow stream of nitrogen. The CLAP analysis of the reaction mixture showed that 1.3% of the unreacted carboxylic acid 3 remained. The reaction mixture was heated to 26 ° C, and 5 ml of DMF was added. The mixture was aged for 2.5 hours. A 1.0 ml aliquot of the reaction mixture was warmed with 5.0 ml of tert-butylamine, and analyzed after evaporation by CLAP: 25 cm RXC8 column Zorbax Dupont at 50 ° C with 1 ml / min flow and detection 220 nm; Isocratic MeCN at 42%, 58% aqueous H3PO4 at 0.1%. This method showed that < 0.05% of acid 3 (estimated by A), and showed > 3% in area of B (> 1 mol% (C0C1) 2).

A B The mixture was concentrated at 10 mbar and at a temperature of 20 to 25 ° C until 5 1 of the solvent had been removed. The typical CLAP profile of the concentrated toluene solution after annealing with t-BuNH2, described above, is as follows: Retention time% in area Identity (min) 2.1 < 0.5% carboxylic acid 3 7.8 < 0.5% Benzyl chloride 11.0 > 99% Cbz-t-butylcarboxamide A 12. 1 NA Toluene 12.7 < 0.5% Diter-butyloxamide B EXAMPLE 3 4-carboxyaldehyde-1-benzyl carbamate of piperidine (5) Materials: N-benzyl carbamate of isonipecotic acid chloride (4) 3.38 kg (12.0 moles) in toluene (p.m. = 281.74) in 5.54 kg DIEA (KF = 18 mg / l) 1.55 kg (15.0 moles) Pd / C at 10% (KF <20 mg / g) 101 g Thioanisole (p.m. = 124.21, d = 1.058) 0.56 g DIEA and thioanisole were added to the solution of (4) in toluene from the previous step, and the catalyst was suspended in this mixture. The mixture was immediately placed in the 18.925 1 autoclave, and hydrogenated at 20 ° C and 2.812 kg / cm2 of H2. After 18 hours, the reaction had captured 70% of the theoretical amount of hydrogen, and the CLAP analysis of an aliquot that was quenched with tert-butylamine indicated that 14.2% remained in the area of acid chloride 2. CLAP conditions They were identical as described above. Retention time: 5 = 8.1 min. A second load of catalyst (101 g) and thioanisole (0.54 g) was added to the hydrogenator as a suspension in 1375 ml of toluene. After 23 hours of CLAP analysis of an aliquot that was quenched with tert-butylamine, it indicated that 1.8% remained in the area of acid chloride 2. The mixture was purged with nitrogen, and the precipitated catalyst and DIEA »HC1 were removed by filtration through Solka-floc ™. The filter cake was washed with 10 1 of toluene. The filtrates were transferred through a 10 μl in-line filter to a 50 1 extractor, and washed with 2 x 7.2 1 of 1 M aqueous HCl and 2 x 7.2 1 of water. The mixture was concentrated at 10 mbar, and at a temperature of 25 to 30 ° C until 5 liters of residue remained.

Retention time% in area Identity (min) 2.1 < 2 carboxylic acid 3 6.6 < 1 dimer 21 8.1 > 95 aldehyde 5 The test yield of aldehyde 3 was 94% by CLAP analysis.

EXAMPLE 4 CBZ-espi roindolina (9) Materials: 4-carboaldehyde-1-benzyl carbamate piperidine (5) 1.71 kg (6.89 moles) in toluene solution in 21.4 kg Phenylhydrazine 900ml, 981g (9.15 moles) Trifluoroacetic acid (TFA) 2.201, 3.26kg (28.6 moles) NaBH * 300g, (7.93 moles) Toluene 34.4 kg MeCN 7.0 1 MeOH 7.0 1 The crude solution of the aldehyde 5 from the previous step was transferred through a 10 μl in-line filter to a 100 1 reactor equipped with Teflon-coated copper coils for cooling or heating and a mechanical stirrer. Toluene (34.4 kg) and MeCN (7 1) were added, and the resulting solution was cooled to 0 ° C. The phenylhydrazine was added in portions, and the temperature was maintained at -1 to 3 ° C, while nitrogen was continuously bubbled through the reaction mixture. Phenylhydrazine was added until the TLC and CLAP analysis indicated complete consumption of aldehyde 5 and the appearance of a slight excess (< 5%) of phenylhydrazine. TLC conditions: Silica, Kieselgel G60 F254 0.25 mm E. Merck; diethyl ether / pentane (4/1); and development agent, 0.5% serum sulfate, 14% ammonium molybdate in 10% aqueous sulfuric acid, and subsequent heating; Rf: aldehyde 5 = 0.52, phenylhydrazone 7 = 0.61, phenylhydrazine 6 = 0.21. CLAP conditions: Zorbax Dupont 25 cm RXC8 column at 30 ° C with flow at 1.0 ml / min, and detection at 254 nm; Gradient program: Time (min) acetonitrile: water 0 57:43 10 65:35 15 75:25 18 75:25 retention times: phenylhydrazine 6 = 4.5 min, toluene = 7.2 min, phenylhydrazone 7 = 11.4 min. The reaction mixture was aged for 30 minutes at 0 ° C at 2 ° C, and TFA was added maintaining the temperature between 2 and 7 ° C. The reaction mixture was heated at 50 ° C for 30 minutes, and kept for 17 hours. The nitrogen mist was stopped through the reaction mixture, and a slow stream of nitrogen was maintained during the reaction mixture. During the first hour at 5 ° C, the color gradually darkened to a dark green, and a relatively small amount of a crystalline white precipitate formed (ammonium trifluoroacetate). After 17 hours, the CLAP analysis (identical conditions as described above) indicated that the reaction mixture contained 91.6% area of indolenin 8, and that 1.5% of unreacted phenylhydrazone remained. Aging the mixture for longer periods did not increase the test yield of indolenin 8. The reaction mixture was cooled to 12 ° C, and 7.0 1 of MeOH was added. NaBH <was added; 4 in small portions (< 20 g), keeping the temperature below 15 ° C. The addition took 30 minutes. Moderate hydrogen production was observed during the addition, but was easily controlled, and virtually no foam formation occurred. Near the end of the addition, the color changed rapidly from green to brown, and then to bright orange. A small amount (< 200 ml) of a heavier phase had separated (possibly aqueous salts). The CLAP analysis (identical conditions as described above) indicated that all indolenine 8 had been consumed (90.4% in CBZ-indoline 9 area); retention times: indolenin 8 = 7.5 min, indoline 9 = 8.2 min. TLC: ethyl ether as a solvent, ceric sulfate dye-ammonium molybdate or 1% anisaldehyde dye; Retention factors: indolenine 8 = 0.18, CBZ-indoline 9 = 0.33. The color change from green to orange corresponds very closely to the end point of the reaction. The amount of NaBH * needed to conclude the reaction depends largely on the temperature and rate of addition of NaBH *, but the yield and quality of the product are virtually unaffected, provided the reaction is over. The reaction mixture was cooled to 5 ° C over a period of 30 minutes. Then, 8 1 of 3% aqueous NH 3 OH was added to bring the pH of the aqueous phase to 7.4, and the mixture was stirred and allowed to settle. The temperature rose to 15 ° C. The lower aqueous phase was turbid yellow colored. The organic phase was washed with 4 1 of 3% aqueous NH 4 OH, 2 x 4 1 of water, and 2 x 4 1 of brine. The weight of the organic phase after washing was 53.5 kg, and the test yield was 94%. The washed toluene solution was combined with the washed organic phases of two other reactions processed in the same way. The total amount of aldehyde used in the three reactions was 5.06 kg (20.5 moles). The total weight of CBZ-indoline 9 tested in the combined organic phases was 5.91 kg (18.3 moles, 90% test yield). The combined organic phases were dried with 5 kg of sodium sulfate, treated with 250 g of Darco G60 carbon for 30 minutes, and filtered through Solka-floc ™. The filtrates were concentrated in vacuo at 10 bars at < 25 ° C until the residue was close to drying. The change of solvent was concluded very slowly by extracting 30% IPAC, and reconcentrating up to 14 1 to 200 mbar at 50-60 ° C. The mixture was heated to reflux to obtain a clear homogenous, bright orange solution. The 1 H NMR analysis indicated that the solution contained approximately 6 mole% residual toluene after solvent exchange. The solution was cooled to 68 ° C, and seeded with 4 g of crystalline CBZ-indoline 9. The solution was allowed to gradually cool to 26 ° C for 6 hours, and was aged for 9 hours at 20-26 ° C. The suspension was cooled to 2 ° C for 1 hour, and aged at 2 ° C for 1 hour. The product was isolated by filtration, and the filter cake was washed with 2 x 2 1 IPAC at 5 ° C, and 2 x 2 1 MTBE at 5'C. The product was dried in the vacuum oven at 30 ° C under a nitrogen charge to give 4.37 kg (74%) of the title compound 9 as a light brown crystalline powder. The CLAP analysis of the product indicated 99.5% in area of purity. The stock solution (11 1) and the washings contained 1.15 kg (19%) of the additional product 9, and approximately 3% Cbz-phenylhydrazide of isonipecotic acid (retention time = 4.8 min).

EXAMPLE 5 CBZ-espi roindol ina methanesulfonamide (1) Mate rials: CBZ-espi roindolina (9) 1.69 kg (5.23 moles) Methanesulfonyl chloride 599 g (5.23 moles) Et3N (KF = 151) 635 g (6.27 moles) THF (KF = 41) 12 1 A 22 1 flask was charged with solid CBZ-spiro roindolin 9, and then 11.5 1 THF and Et3N were transferred from the flask through a 10 μ filter in line. The resulting homogeneous solution was cooled to 0 ° C. A 1 1 dropping funnel was charged with the methanesulfonyl chloride and 500 ml of THF. The solution of the MsCl in THF was added to the reaction mixture maintaining the temperature between 0 and 4 ° C. The addition took 5 hours, and was exothermic. A white precipitate, probably triethylammonium hydrochloride, was formed during the addition. The CLAP analysis indicated that the reaction ended at the end of the addition (9 was undetectable). CLAP conditions: Zorbax Dupont 25 cm RXC8 column with 1.5 ml / min flow and detection at 254 nm. Gradient program: Time (min) Hs P0 aqueous at 0.1%: MeCN 0 70:30 3 70:30 12 20:80 25 20:80 Retention times: 9 = 7.6 min, 1 = 13.6 min.

After the addition was complete, the reaction mixture was heated to 18 ° C and aged for 16 hours. There was no change in the appearance of the reaction mixture and the CLAP profile between the end of the addition and after 16 hours of aging. The reaction mixture was transferred slowly for 1 hour in a vigorously stirred solution of 30 1 of water and 200 ml of aqueous HCl at 37% in a 50-ml flask. 1. The temperature in the 50 1 flask was raised from 22 to 28 ° C.

The product separated as a pale tan gummy solid which changed to a granular solid. The aqueous suspension was cooled to 22 ° C and aged for 1 hour. The suspension was filtered, and the filter cake was washed with 2 x 4 1 MeOH / water (50/50). The CLAP analysis indicated that < 0.1% of CBZ-espi roindoline methanesulfonamide 1 was in the stock solution. The filter cake was washed with 4 1 MeOH / water (50/50) to which 50 ml of 28% aqueous NH 0 H had been added. The filter cake was washed with 2 x 4 1 MeOH / water (50/50), and the solid was dried in the vacuum oven at 50 ° C under nitrogen loading to give 2.03 kg (97%) of the product 1 of the title as a white powder. The CLAP analysis of solids indicated 93.7% in area of 1.

EXAMPLE 6 Optional procedure for the isolation of intermediary CBZ-espi roindolenin (8) Materials: 4-carboxyaldehyde-l-benzyl 12.37 g (0.050 moles) piperidine carbamate (5) Phenylhydrazine 5.41 g (0.050 moles) Roacetic t-fl uo fl uid acid (TFA) 11.56 ml, 17.10 g (0.150 mol) Methylene chloride 500 ml The CBZ-aldehyde 5 was dissolved in dichloromethane in a flask of 11 equipped with magnetic stirring bar coated with Teflon. The resulting solution was cooled to 0 ° C. Phenylhydrazine was added via a heavy syringe for 5 minutes, and the temperature was maintained at -1 to 3 ° C, while nitrogen was continuously bubbled through the reaction mixture. The analysis of TLC and CLAP indicated the complete consumption of CBZ-aldehyde 5, and the appearance of a slight excess (< 2%) of phenylhydrazine. TLC conditions: Silica, Kieselgel G60 F254 0.25 mm E. Merck; diethyl ether / pentane (4/1); and development agent, 0.5% ceric sulfate, 14% ammonium molybdate in 10% aqueous sulfuric acid, and subsequent heating; Rf: aldehyde 5 = 0.52, phenylhydrazone 7 = 0.61, phenylhydrazine 6 = 0. 21. CLAP conditions: Zorbax Dupont 25 cm RXC8 column at 30 ° C with flow of 1.0 ml / min, and detection at 254 nm; Gradient program: Time (min) acetonitrile: water 0 57:43 10 65:35 15 75:25 18 75:25 retention times: phenylhydrazine 6 = 4.5 min, toluene = 7.2 min, phenylhydrazone 7 = 11.4 min.

The reaction mixture was aged for 10 minutes at 0 to 2 ° C, and TFA was added by syringe keeping the temperature between 2 and 7 ° C. The reaction mixture was heated at 35 ° C for 30 minutes, and it was maintained for 17 hours. The nitrogen mist through the reaction mixture was stopped, and a slow stream of nitrogen was maintained on the reaction mixture. During the first hour at 35 ° C, the color gradually darkened to a pink color and then to a deep green color, and a relatively small amount of a white crystalline precipitate (ammonium trifluoroacetate) was formed. After aging for 17 hours, the CLAP analysis (identical conditions as described above) indicated that the reaction mixture contained 93% in indolenin 8 area and that it remained <0.5% unreacted phenylhydrazone. Aging the mixture for longer periods did not increase the test yield of indolenin 8. The reaction mixture was cooled to 10 ° C, and a mixture containing 60 ml of ammonium hydroxide at 28 ° C was added with vigorous stirring. 30%, 90 ml of water and 150 g of crushed ice. The color of the mixture changed to a salmon color. The organic phase was separated and washed twice with 400 ml of water and then with 100 ml of saturated aqueous NaCl. The organic phase was dried over magnesium sulfate, and filtered through a plug of 5 g of silica. The filtrate was evaporated to give 15.84 g (99%) of indolenine 8 as a pale orange oil.

EXAMPLE 7 Procedure for the preparation of CBZ-Spiroindoline-methanesulfonamide (1) without isolation of intermediate CBZ- Espi roindolina (9) Step 1: CBZ-Espi roindoline (9) Materials: Piperidine-4-carboxaldehyde-1-benzyl carbamate 49.5 g (020 moles) Phenylhydrazine (Aldrich) 23.7 g (0.22 moles) Trifluoroacetic acid (TFA) 75.4 g (0.66 moles) Toluene (KF <250 mg / l 654 ml MeCN (KF <250 mg / l 13.3 ml NaBH4 11.3 g, (0.30 moles) Toluene 20 ml MeOH 50 ml A 2% (by volume) solution of MeCN in toluene was made using 654 ml of toluene and 1.3 ml of MeCN. In a 2-liter 3-necked flask equipped with a mechanical stirrer, 617 ml of the above solution was degassed by passing a fine stream of nitrogen through the solution for 5 minutes. Phenylhydrazine and TFA were added to the mixture while degassing. The CBZ-aldehyde 5 was dissolved in the rest of the solution prepared above (50 ml) and degassed by bubbling nitrogen through the solution while it was in the addition funnel. The solution in the flask was heated to 35 ° C and the adehide solution was slowly added to the phenylhydrazine-TFA for 2 hours. The mixture was aged at 35 ° C for 16 hours. CLAR conditions: a Dupont Zorbax RXC8 column of 25 cm at 50 ° C with 1 ml / min flow and detection at 220 nm; 55% isocratic MeCN, 45% aqueous H3PO4 at 0.1%. Typical CLAR profile after 16 hours of aging: Retention time (min)% of area Identity 1.6 0.1-0.5 phenylhydrazine 6 4.1 < 0.1 dimer 21 4.7 < 0.1 aldehyde 5 5.0 NA spiroindoline 9 6.3 NA toluene 6.9 97 spiroinindole 20.3 < 0.2 pennylhydrazone 7 2-3 tot. other impurities < 0.2% c / u The mixture was cooled to -10 ° C and MeOH was added. A suspension of sodium borohydride in 20 ml of toluene was added in small portions (1 ml) for 30 minutes taking care that the temperature did not exceed -2 ° C.

Area% Identity 0.1-1 phenylhydrazine 6 85-90 CBZ-espi roindoline 9 < 0.1 CBZ-espi roindolenina 8 10-15 tot. Other impurities (< 3% c / u) The temperature was raised to 10 ° C for 1 hour, and 6% aqueous ammonia (200 ml) was added. The mixture was stirred for 10 minutes, allowed to stand for another 10 minutes and the lower aqueous phase was separated. Acetonitrile (20 ml) and MeOH (20 ml) were added to the organic phase and washed with 150 ml of 15% brine. It was found that the organic phase contained a test yield of 92% CBZ-spiro roindoline 9.

Step 2: CBZ-Spiroindoline-methanesulfonamine (1) Materials: CBZ-Espi roindoline (9 = (MW = 322.51) (0.184 moles) Methanesulfonyl chloride 21.1 g (0.184 moles) DIEA (KF = 150 mg / l) 29.7 g, 40.1 ml (0.230 mol) THF (KF = 41 mg / l) 150 ml The crude solution of the CBZ-spiro-roindoline solution 9 from step 1 above was concentrated in a flask of 3 necks of 1 1 (60-70 ° C, 150-200 Torr) until there were 250 grams of residue. The THF and DIEA were added, and the resulting homogeneous solution was cooled to 0 ° C. A 125 ml dropping funnel was charged with methanesulfonyl chloride and 50 ml of THF. The solution of MsCl in THF was added for 2 hours to the reaction mixture maintaining the temperature between 0 and 4 ° C and the mixture was aged for 2 hours at 5-8 ° C. The addition was slightly exothermic. A white precipitate, presumably DIEA hydrochloride, was formed during the addition. The conditions of CLAR were the same as before. The CLAR analysis indicated that the reaction was completed 1 hour after the end of the addition (9 was undetectable) and the test yield was 94% from 9. Retention time: 1 = 7.8 min. Typical CLAR profile of reaction mixture after 2 hours of aging: % Identity area < 0.1 CBZ-espi roindoline 9 90-92 CBZ-sulfonamide 1 8-10 tot. other impurities (< 2% each) The mixture was heated to 20 ° C and 200 ml of Aqueous HCl at 1 M. The mixture was heated to 50 ° C, and the aqueous phase was separated. The organic phase was washed sequentially with 100 ml of water, 100 ml of 5% aqueous sodium bicarbonate and 100 ml of water. The organic phase was transferred to a 1-liter 3-necked flask equipped for mechanical stirring and distillation. The mixture (approximately 400 ml) was distilled at atmospheric pressure until 150 ml of distilled product had been collected. The temperature of the distillation head reached 107 ° C; the temperature of the vessel was 110 ° C. The distillation was continued with continuous addition of n-propanol at a rate such that a constant volume (about 350 ml) was maintained in the vessel. The distillation was stopped when 525 ml of n-PrOH had been added and a total of 800 ml of distilled product had been collected. The temperature of both the distillation head and the vessel was raised from 94 ° C to 98 ° C during solvent exchange. Toluene and n-PrOH form an azeotrope that boils at 97.2 ° C composed of 47.5% toluene and 52.5% n-PrOH. The mixture was allowed to cool gradually to 20 ° C for 3 hours and aged for 12 hours. The stock solution was found to contain 2% toluene and 4 mg / ml sulfonamide. The solubility of the sulfonamide in various mixtures of toluene and n-PrOH has been determined by HPLC test:% of Toluene in n-PrOH Solubility of 1 in mg / ml 0 2.36 5 3.02 10 4.23 20 7.51 25 10.3 The crystalline suspension is leaked and washed with 3 x 100 ml of n-PrOH. The product was dried in a vacuum oven to 50 ° C with a nitrogen purge for 16 hours to produce 65. 5 g (82% aldehyde 5) of 6 as a tan solid with a purity of 93.5% by weight. Typical solid CLAR profile:% Identity Area < 0.1 CBZ-Espi roindolin 9 > 99 CBZ-sulfonamide 1 1 tot. other impurities (< 0.2% each) For further purification, 40.0 g of the sulfonamide sample crystallized with n-Pr-OH were dissolved in 134 ml of EtOAc at 60 ° C and treated with 8.0 g of Darco G-60 carbon for 1 hour at 60 * C. After the addition of 2.0 g of Solkafloe ™, the suspension was filtered through a 4.0 g pad of Solkafloe ™, and the pad was washed with 90 ml of EtOAc at 60 ° C. Before the carbon addition, the solution had a brown color. The filtration proceeded well without covering to give a filtered yellow gold product. The filtrate was distilled at atmospheric pressure in a 500 ml flask (vessel temperature 80-85 ° C) until 100 g of residue remained. The solution was allowed to cool to 35 ° C for 3 hours. During a period of 1 hour, 116 ml of hexane cycle was added with good agitation at 35 ° C. The mixture was cooled to 20 ° C for 1 hour and aged at 20 ° C for 12 hours. At 35 ° C much of the sulfonamide had crystallized and the mixture had thickened. The addition of cyclohexane at 20 ° C made agitation difficult. After the aging period, the supernatant was found to contain 2.5 mg 1 / g. The crystalline suspension was filtered and the cake was washed with 77 ml of 2: 1 cyclohexane-EtOAc and 2 x 77 ml of cyclohexane. The product was dried in a vacuum oven at 50 ° C with nitrogen purge for 16 hours to give 34.2 g of 1 (MW = 400.3) as a white crystalline solid (85% recovery of the crude 1, 70% of 5 with a purity of> 99.9 percent by weight).

EXAMPLE 8 Salt of spiroindoline-methanesulfonamide hydrochloride (la) Materials: CBZ-Spiroindolinesulfonamide (1) 941 g (2.35 moles) Pearlman Catalyst Pd (0H) 2 at 20% / C 188 g THF 8 1 MeOH 7 1 The catalyst was suspended in 7 1 MeOH and transferred to the autoclave of 18.9 1 followed by the solution of 1 in 8 1 THF. The mixture was hydrogenated at 25 ° C to 5.6 kg2 of H2. After 2.5 hours, the temperature was raised to 35 ° C for 30 minutes. The CLAR analysis indicated complete consumption of Cbz-spiroindoline-methanesulfonamide. CLAR conditions: with a Zorbax RXC8 25cm dupont with 1.5 ml / min flow and detection at 254 nm.

GRADIENT SCHEME: Time (mi n) H3 P04: 0.1% aqueous MeCN 0 70: 30 3 70. 30 12 20: 80 25 20: 80 Retention times: Espi roindoline = 7.6 min, Cbz -spyroindoline-methanesulfonamide = 13.6 min. The mixture was purged with nitrogen and the catalyst was removed by filtration through Solka-floc ™ while heating. The catalyst was washed with 4 1 THF and 2 1 MeOH. The pale yellow filtered products were concentrated to a thick oil at 10 bars and < 25ßC. The solvent change was completed by purging slowly in 15 1 of EtOAc and reconcentrating to dryness. The residue solidified to a hard whitish mass. MeOH (1.5 1) was added and the mixture was heated to 70 ° C to give a homogeneous solution. Although the solution was at 70 ° C, 10.5 1 EtOac was added at 20 ° C. The temperature dropped to 40 ° C, and the mixture remained homogeneous. Subsequent experiments suggested that it is more convenient to solvent change the MeOH-THF-filtered products to MeOH, concentrate to the desired volume and then add the EtOAc. This prevents the solidification of the residue under the concentration of the EtOAc solution. Diluted hydrochloric acid with approximately equal volume of nitrogen was passed to the solution. The temperature was raised to 60 ° C over the course of 15 minutes, and a white precipitate of the hydrochloride salt formed. Diluting the HCl with nitrogen only prevents the reaction mixture from being retrosulated and may not be necessary. The mixture was cooled in an ice bath and the addition of hydrochloric acid was continued for one hour. The temperature fell gradually to 20 ° C. The suspension was aged for 2 hours while the temperature was reduced to 10 ° C. The crystalline product was isolated by filtration, and the filter cake was washed with 3 1 EtOAc. It was dried in the vacuum oven at 35 ° C to give 1.18 kg (86%) of the title product as a whitish crystalline solid of% area purity. 99.5 by CLAR analysis. The conditions of CLAR were: a Duppont Zorbax RXC8 25 cm column with a flow of 1.5 ml / min and the detection at 230 n; 35% isocratic MeCN, 65% aqueous 0.1% ammonium acetate. Retention time: la = 5.4 min.

EXAMPLE 9 Spiroindoline-etansulfonamide (free base form) Ib) An aliquot of 250 ml of the filtrate from the hydrogenolysis of Cbz- containing 4.67 g of Ib (free base) was concentrated to approximately 10 ml. The residue was dissolved in 20 ml of EtOAc and the solution re-concentrated to approximately 10 ml. This was repeated one more time, and 10 mL of EtOAc was added to the residue. A crystalline precipitate began to form. MTBE (20 ml) was added in one portion. The additional crystalline solid was precipitated, but the supernatant still contained a substantial amount of dissolved product that did not precipitate during rest. Hexanes (70 ml) were added dropwise during 2 hours to the mixture with vigorous stirring. Slow addition of the hexanes is necessary to avoid separation of the amine. The stirred mixture was aged for 1 hour and filtered. The filter cake was washed with 20 ml of 1: 1 MTBE-hexanes and then with 20 ml of hexanes. The product was dried under a stream of nitrogen to give 3.86 g (82%) of the Ib-free amine as a whitish crystalline solid of% area purity. 99.5. The CLAR conditions were: Dupont Zorbax RXC8 25 cm column with a flow of 1.5 ml / min and detection at 230 nm; MechaN at 35% isocratic, 65% aqueous ammonium acetate at 0.1%. The retention time: Ib = 5.4 min.

EXAMPLE 10A Spiroindoline-etansulfonamide (free base form) (b) Materials: CBZ-espi roindoline-sulfonamide (1) 833.5gr (2.08 moles) Pd (0H) 2 / C (20% by weight of Pd (0H) 2) 124.5 (15%) THF 6.5 1 MeOH 19.5 1 NH4OH (conc) 60 ml Hydrogenation was performed 3 times due to equipment limitations; this procedure refers to a single operation. The CBZ-espi roindoline-sulfonamide 1 was dissolved in THF (6.5 1, KF = 53 μg / μl) and then MeOH (KF = 18 μg / gml, 4 1 was added followed by the addition of the catalyst and the suspension was transferred to a Autoclave 18.9 1. The rest of the MeOH (2.5 1) was used for rinsing.The mixture was heated at 40 ° C to 3.5 k / cm2 for 24 hours.The catalyst loading and the reaction time are a function of the purity of the starting material 1. This material was unique requiring> 15% catalyst and a long reaction time The purest lots of spiroindoline required only 5% catalyst and a reaction time of 4 to 6 hours. To complete (<0.1 A% 1 by LC) the mixture was filtered through Solka Floc ™ and the carbon cake was washed with MeOH (13 1) containing NH 4 OH (0.5%, 60 ml) The combined filtrates ( the test shows 1587 g of spiroindoline-amine Ib) were concentrated in vacuo and the resulting solids were divided between 40 1 (of toluene : THF (3: 1) and 0.5N NaOH (18 1). Although the layers separated easily, a strong precipitate could be seen in the aqueous layer. The aqueous suspension was therefore extracted with CH2C1-2 (15 1). The aqueous and organic layer separated slowly. Prior to the addition of CH2Cl2, THF was added to the aqueous layer together with sufficient NaCl to saturate the layer. However, the dissolution of the product was not achieved, which necessitated the use of CH2CI2.

The combined toluene layers THF and CH2Cl2 were combined and concentrated in the intermittent concentrator.

The residue was flushed with 7 1 CH 3 CN. Finally, 10 1 of CH 3 CN was added and the solution was allowed to stand overnight under N 2 atmosphere.

EXAMPLE 10B Spiroindoline-etanesulfonamide (free base form) (Ib) Materials: CBZ-espi roindoline-sulfonamide (1) 3kg (7.49 moles) Darco G-60 600 g Ethyl acetate 36 1 Absolute ethanol 189 1 10% Pd / C 450 g Ammonia solution 500 ml Solka Floc ™ 2.5 kg Isopropyl acetate 65 1 A mixture of CBZ-espi roindonila (1) (1 kg) and Darco G-60 (200 g) in ethyl acetate (9 1) was stirred and heated to 60-65 ° C under a nitrogen atmosphere for 8 hours . The Darco was removed by filtration at 60-65 ° C, the solid was washed with hot ethyl acetate (3 1) and the filtrate and the washings were combined. The LC p / p test confirmed negligible loss for Darco. The ethyl acetate solution was evaporated to dryness in vacuo using a 20 1 Buchi apparatus and then jet washed with absolute ethanol (2 x 5 1). This material was then suspended in absolute ethanol (8 1) heated to 65-70 ° C and placed in the autoclave of 20 1. The batch was rinsed in absolute ethanol (11). A 10% suspension of palladium on carbon (75g, 7.5% by weight) in absolute ethanol (750 ml) was then added to the autoclave and rinsed with an additional portion of absolute ethanol (250 ml). The batch was hydrogenated at 65 ° C with vigorous stirring under hydrogen pressure of 2.8 kg / cm2 for 3 hours, a second portion of 10% palladium on charcoal (75 g) the batch was hydrogenated for an additional 2 hours and then sealed overnight. The batch was transferred (still hot, 60-65 ° C) to a Buchi apparatus of 20 1 and degassed under vacuum to remove formic acid by "feeding and purging" absolute ethanol (a total of 18 1). This procedure was repeated 2 more times and the three batches were combined in a 37.8 1 glass lined vessel and the combined batch was degassed again by the addition and distillation (vacuum) of absolute ethanol (2 x 10 1).

Solka floc ™ (0.5 kg) was added to the batch and rinsed with ethanol (10 1). A star filter was loaded with Solkafloe ™ (2 kg) as a suspension in ethanol (20 1). The resulting mixture was heated to 60-65 ° C and then transferred at this temperature through a heated filter using a pump to 2 stainless steel tanks that had been tared. The initial vessel, filter, pump and liners were rinsed with a hot mixture (60-65 ° C) of aqueous ammonia (500 ml) in absolute ethanol (25 1). The filtered product and the washes were combined in the two stainless steel tanks. The batch was then transferred to a vessel using an in-line filter containing a 10-micron cartridge, and then concentrated in vacuo to reduce the volume (~ 15 1). The ethanol was replaced by isopropyl acetate by the "feed and purge" of batch volumes 3 x of isopropyl acetate (45 1 in total), while maintaining a batch volume of 15 1. The change of solvent, when completed, contained < 1% residual ethanol by gas chromatography. The batch was diluted to ~ 33 1 by the addition of isopropyl acetate (20 1), and this solution of spiroindoline-amine Ib (1855 kg by liquid chromatography analysis) in isopropyl acetate was used for the next step of the procedure.

EXAMPLE HAS Boc-O-Benzyl Serine-Espi Roindoline (11) Materials Espi Roindoline-Amine (lb) 1587 g (5,966 moles) Amino Acid (10) 1938 g (6.563 moles), CO2H Ph 'NHBOC DCC 1334.5 g (6,563 moles) HOBT 884 G (6,563 moles) CH3CN 25 1 0.5N NaOH 18 1 0.5N HCl 18 1 N «HC03 sat. 18 1 iPrOAc 28 1 Spiroindoline-a ina Ib in CH3CN or iPrOAc: H2? (25 1) at room temperature under N2 was treated in sequence with HOBT (884 g; 1.1 eq) with a solid, DCC (1334.5 g, 1.1 eq) as the molten bath (heating in hot water at 60 ° C for about 1 hour) and finally amino acid 10 (1938 g) as the solid. The mixture was stirred for 3 hours, time after which the heavy precipitation of DCU occurred and the liquid chromatography analysis showed approximately 0.5 A% remaining amine Ib. IPAc (9 1) was added, the suspension was filtered through Solka Floc ™ and the cake was washed with IPAc (19 1). The combined organic solution was washed in sequence with NaOH at 0.5N (18 1), HCl at 0.5N (18 1) and saturated NaHC 3 (18 1). A final wash with water at this point resulted in an emulsion and then was removed. The organic layer was concentrated in vacuo and the residue was dissolved in MeOH or EtOH (final volume of 10 1). The test produced 3026 gr (89%). The use of alternative peptide coupling agents such as carbonyldimidazole or the formation of mixed anhydrides, such as sec-butyl carbonate, gave lower yields of 11 and / or 14 with a high degree of epimerization in the case of the first compound. Other peptide coupling reagents were prohibitively expensive.

EXAMPLE 11B Boc-O-Benzyl Serine-Spiroindoline (11) Materials: Spiroindoline-amine (Ib) 1,855 kg (6.96 moles) Isopropyl Acetate 29 1 Dicyclohexylcarbodiimide (DCC) 1.58 kg (7.65 moles) 1-Hydroxybenzotriazole (HOBt) 1.03 kg (7.62 moles) N-Boc-O-benzyl-D-Serine 2.26 kg (7.65 moles) Aqueous sodium hydroxide at MM 26 1 Aqueous hydrochloric acid at 0.5M 26 1 Sodium Bicarbonate Aqueous Saturated 26 1 Absolute ethanol 50 1 Water (20 1) was added to the stirred solution of the spiroindoline-amine Ib (1855 kg) in isopropyl acetate (33 1) in a reaction vessel. The following chemical compounds were added sequentially at room temperature under a nitrogen atmosphere: DCC (1.58 kg, 1.1 equiv.), HOBt (1.03 kg, 1.1 equiv.) And finally N-Boc-O-benzyl-D-Serine ( 2.26 kg, 1.1 equiv.). The reagents were rinsed with isopropyl acetate (7 1). The batch was stirred at room temperature under nitrogen atmosphere for 5 hours when liquid chromatography showed that the product / starting material ratio was 99.4 / 0.6. The mixture was then filtered through a star filter using cloth and cardboard alone and using a pump to pump into another container. The container from which it was sent was rinsed with isopropyl acetate (22 1) and this was used to rinse the filter, the pump and the lines in the receiving container. The mixture of two phases in the vessel was stirred for 10 minutes and allowed to settle for 15 minutes. The lower aqueous layer was separated and the organic solution was allowed to stand at room temperature overnight. The next day, the organic solution was washed with an aqueous solution of sodium hydroxide at 1M (261) and then aqueous hydrochloric acid at 0.5M (26 1) and finally saturated aqueous sodium bicarbonate (26 1). Liquid chromatography analysis gave a test yield of 3,787 kg, overall yield of 93% of 7.49 moles (3 kg) of starting CBZ-spiro roindoline (1). The batch was concentrated under vacuum (internal temperature = 13-15 ° C, jacket temperature = 40 ° C, Vacuum = 29") at low volume (~ 15 1) and the solvent was changed to ethanol" feeding and purging "ethanol (50 1) while keeping the volume at ~ 15 1. Gas chromatography showed <1% remaining isopropyl acetate. The solution was used for the next stage of the process.

EXAMPLE 12A O-Benzyl Serine-Espi Roindoline (free base form) (12) Materials: Boc-O-Benzyl Serine-Spiroindoline (11) 3026 g (5.57 moles) Methanesulfonic Acid (MsOH) 1.16 1 (17.9 moles) MeOH 10 1 iPrOAc 24 1 0.5N NaOH 35 1 Boc-O-benzyl serine-spiroindoline 11 in 10 1 of MeOh (or EtOH) was treated with net MsOH (1.16 1) added for approximately 30-40 minutes (initial temperature of 16- ° C final temperature of 28 ° C). The dark red solution was aged overnight under N2. The mixture was then pumped into a 100 1 extractor containing 24 1 iPrOAc and 35 1 0.5 N NaOH The pH of the aqueous layer was 7. NaOH (6M) was added until pH > 10.5. As the pH increased the color changed from red to yellow. The layers were separated and the organic layer (241) was shown by NMR containing 13 mol% MeOH in iPrOAc [5% by volume]. The liquid chromatography test was 2. 48 kg.

EXAMPLE 12B O-Benzyl Serine-Spir roindolin (free base form) (12) Materials: Boc-O-Benzyl Serine-Spiroidoline (11) 3,787 kg (6.96 moles) Methanesulfonic Acid 2,006 kg (20.87 moles) Isopropyl Acetate 38 1 Aqueous Sodium Hydroxide at IM 16 1 50% Aqueous Sodium Hydroxide 1.6 1 Methanesulfonic acid (2,006 kg, 1355 1, ~ 3 equiv.) Was added to the stirred solution of Boc-O-benzyl serine-spiro-roindoline 811) (3.787 kg) in ethanol (total volume of ~15 1 in a reaction vessel. The batch was heated to 35-40 ° C. After 7 hours, liquid chromatography showed the absence of starting material and the reaction was allowed to cool to room temperature overnight.The next day water was added (44 1) The batch was stirred at ~ 5 °, stirred for 30 minutes and then filtered through an in-line filter (loaded with a 10 μm cartridge) into a reservoir, the batch was then retrosulated to the vessel. .

A water rinse (10 1) was used to rinse the container and lines to the reservoir and then used to rinse the container again. Isopropyl acetate (381) was added followed by aqueous sodium hydroxide to IM (16 1). The batch was cooled to 10-15 ° C, the pH of the lower aqueous layer was confirmed as ~7 and a 50% aqueous sodium hydroxide solution (0.6) was added (pH> 10). The batch was stirred at 10-15 ° C for 25 minutes and then allowed to settle for 10-15 minutes. The lower aqueous layer was separated (78.1 kg). The LC test indicated 28.4 g of 12 (0.85% theory) contained in the aqueous solutions. The volume of the organic solution = 51 1. The liquid chromatography test indicated 3.057 kg, 92% overall yield from 3 kg, 7.49 moles of CBZ-espi roindoline-sulfonamide (1). This solution was used for the next stage.

EXAMPLE 13A Boc-Aminoisobuti ril-O-Benzyl Serine-Espi Roindoline (14) Mate riales: Espi roindolina-amina (12) 2481 g (5.57 moles) Amino acid peptide (13) 1347.1 g (6.16 moles) DCC 1266.7 g (6.16 moles) HOBT 827 g (6. 16 moles) IPAc 52 1 H20 37 1 0.5N NaOH 36 1 0.5N HCl 36 1 Sat. NaHC? 3 36 1 The solution of the amine in 12 in IPAc was diluted to a total volume of 39 1 with IPAc and 37 1 of H2O was added. The biphasic mixture was then treated in sequence with HOBT (827 g) as a solid, DCC (1266.7 g) as a molten bath, and amino acid 13 at room temperature under nitrogen. The reaction mixture was stirred for 2 hours, after which the liquid chromatography analysis indicated the disappearance of starting material 12 (< 0.3 A%). The mixture was filtered through Solka Floc ™ and the solids were washed with 13 1 of IPAc. The material was stored at this point as a two-phase mixture at night.

The mixture was transferred to a 100 1 extractor, the aqueous layer was separated and the organic layer was washed successively with 36 1 of NaOH at 0.5N, HCl at 0.5N and saturated NaHCO 3. The test gave 3160 g (81% from spiroindoline + 5% for volume measurement error). The solution was concentrated to a small volume and flushed with ethanol (2 x 4 1). If desired, intermediate compound 14 can be isolated by adding water to crystallize it. The use of alternative peptide coupling agents such as carbonyldiimidazole or formation of mixed anhydrides, such as sec-butyl carbonate gave the lower yields of 14 with a high degree of epimerization. Other peptide coupling reagents were prohibitively expensive.

EXAMPLE 13B Boc-Aminoisobuti ril-O-Benzyl Serine-Espi Roindoline (14) Materials: Spiroindoline-amine (12) 3.057 kg (6.89 mol) Dicyclohexylcarbodiimide (DCC) 1.56 kg (7.56 mol) 1-Hydroxybenzotriazole (HOBt) 1.02 kg (7.55 mol) 2-aminoisobutyl acid (13) 1.54 kg (7.58 mol) Isopropyl Acetate 32 L Aqueous Sodium Hydroxide at IM 38 L Aqueous Hydrochloric Acid at 0.5 M 38 L Saturated Aqueous Sodium Bicarbonate 38 L Absolute Ethanol 45 L Water (49 L) was added to the stirred solution of spiroindoline-amine 12 (3057 kg) in isopropyl acetate (total volume ~ 51 L) in a reaction vessel at room temperature under a nitrogen atmosphere. The following chemical compounds were added sequentially: DCC (1.56 kg, ~ 1.1 equiv.), HOBt (1.02 kg, ~ 1.1 equivs-.) And finally N-Boc-2-aminoisobutyl acid 13 (1.54 kg, ~ 1.1 equivs.). The mixture was stirred vigorously at room temperature for 2 hours when liquid chromatography showed that the reaction was complete. The mixture was filtered to another vessel through a Star filter using a pump. Isopropyl acetate (22 L) was used to rinse the container, filter, pump and lines to the receiving container. The two-phase mixture was then stirred for 5 minutes and the layers were allowed to separate. The lower aqueous layer was separated without incident (weight of aqueous solutions = 51.1 kg). The organic solution was then washed sequentially with aqueous sodium hydroxide to IM (38 L), aqueous hydrochloric acid to 0.5M (38 L) and finally saturated aqueous sodium bicarbonate (38 L) without incident. The organic solution was then transferred using a pump through an in-line filter (containing an IOμ cartridge) to another vessel for the change of solvent to ethanol. The vessel was rinsed with isopropyl acetate (10 L) and this was used to rinse the pump, filter and lines to the receiving vessel. The filtrate and the washings were combined. Total volume = 75 1 (by immersion rod). The liquid chromatography test gave 4.395 kg of Boc-aminoisobutyryl-O-benzyl serine espi roindoline (14), that is, 93% of the total of 7.49 moles of starting CBZ-espi roindoline-sulfonamide (1). The batch was concentrated in vacuo to a lower volume (~15 L) and the isopropyl acetate was changed to ethanol "feeding and purging" with absolute ethanol (45 L total). At the end of the solvent change, the gas chromatography showed < 1% remaining isopropyl acetate. This solution (25 L) containing 4395 kg of 14 was used for the next step. If desired, intermediate compound 14 can be isolated by adding water to crystallize it.

EXAMPLE 14A Ami noisobut i ril-O-bencilse quarrel -spyroindoline (15) Materials: Boc espi roindolina (14) 3160 g (5.03 moles) Methanesulfonic acid (MsOH) 979 mL (15.1 moles) EtOH 6.2 L H20 30 L INN NaOH 11 L EtOAc 26 L Charcoal activated Darco 60 1 Kg The bovine spiroindoline 14 was dissolved in 6.2 L of EtOH and treated with MsOH (979 mL). The temperature rose from 20 to 30 ° C and the reaction was allowed to proceed overnight. After 12 hours at 20 ° C there was still 15% A of starting material left, so the mixture was heated at 35 ° C for 6 hours. Upon completion (<0.1% A 14) the reaction was cooled to 20 < C and 30 L of H2O were added and the solution was filtered through a glass funnel with a polypropylene filter to filter residual DCU The mixture was transferred to a 100 L extractor and 26 L of EtOAc was added. aqueous layer was basified by the addition of cooled NaOH to IN (11 L) and 1 L of 50% NaOH.Addition of ice was required to maintain the temperature below 14 ° C. Higher temperatures were obtained in emulsion problems The organic layer was distilled at 50 ° C to about 53.34 cm Hg to KF <1000 μg / mL.The lower KFs resulted in more efficient carbon treatments and better recovery in the salt formation step. of 160 μg / mL were achieved at the scale of 700 grams The solution was diluted with ethyl acetate to a total volume of 31 L (liquid chromatography test gave 2.40 kg) Activated carbon (Darco G-60) was added and the mixture was stirred for 24 hours. it was filtered through Solka Floc ™ and the filter cake was washed with ethyl acetate (16 L), the test giving 2.34 kg.

EXAMPLE 14B Aminoisobutyryl-O-benzyl serine-espi roindoline (15) Materials: Boc espi roindolina (14) 4.395 kg (6.99 mol) Methanesulfonic acid 2.017 kg (20.99 mol) Ethyl acetate 185 L Aqueous sodium hydroxide at M 16 L Aqueous sodium hydroxide at 50% 2.6 L Darco G-60 900 g Solka Floc ™ 2.5 kg Methanesulfonic acid (2.017 kg, 1.36 L, ~ 3 equiv.) Was added to the stirred solution of Boc Spirindoline 14 (4395 kg) in ethanol (total volume ~25 L) in a reaction vessel at room temperature. The batch was heated to 35-40 ° C, and stirred overnight. The next day, the batch contained ~ 1.1 A% of the starting material and in this way the reaction was continued for an additional 4 hours, then the liquid chromatography showed a product / starting material ratio of 99.6 / 0.4. The batch was concentrated in vacuo to a volume of ~15 L and then diluted with water (44 L). The batch was cooled to 5 ° C, stirred for 30 minutes and then filtered through a Sparkler in-line filter (containing a 10 μ cartridge) using a pump to pump to another vessel in order to remove a small amount. of residual DCU. The container, the pump and the filter as well as the lines were rinsed with water (10 L), and this was added to the container. Ethyl acetate (36 L) was added and the vessel and the stirred mixture were cooled to 10 ° C. A cold aqueous solution of sodium hydroxide at 50 ° C (5-10 ° C) (16 L) and a cold 50% aqueous sodium hydroxide solution (5-10 ° C) (2.6 L) were added at 10 ° C. and the temperature rose to 14 * C. The resulting mixture was stirred for 15 minutes at < 14 ° C and then the lower aqueous layer was separated. The batch was concentrated in vacuo to a volume of ~20 L and then a mixture of ethyl acetate (35 L) and ethanol (5 L) was fed while maintaining the volume at "20 L. At the end of the distillation, the KF was 9160 mgml-1, the batch was solvent changed to ethyl acetate "feeding and purging" ethyl acetate (total of 40 L.) At the end of this distillation, KF was 446 mgml-1. with ethyl acetate (10 L) Darco G-60 (900 g) was added to the cloudy mixture, which was rinsed with ethyl acetate (6 L) The mixture was stirred at room temperature overnight. Solka Floc ™ (0.5 kg) was added to the agitated batch in the vessel and then the Solka Floc ™ (2.0 Kg) was stirred in little ethyl acetate and loaded onto a Star filter.Excess solvent was pumped away to through a Sparkler in-line filter that contained an IOμ cartridge.The suspension was transferred from the container through a filter using a pump Ba and then through another filter to 2 x 40 L stainless steel tanks. Visual inspection showed that the solutions were clear and clean. The vessel was rinsed with ethyl acetate (22 L) and this was used to rinse through the route previously delineated to the stainless steel cans. The contents of both cans were transferred to the reaction vessel and the solution was mixed uniformly. The batch (58 L) had a KF of 2950 mgml -1"and in this way it was again dried by concentrating in vacuo at a volume of 20-25 L. The batch was diluted to a volume of 46 L (immersion rod) by the addition of ethyl acetate (25 L) The KF was 363 mgml "1. The batch was diluted to a volume of 62 L by the addition of ethyl acetate (17 L) and used for the final stage of the process.

EXAMPLE 15A Me 3.4'-piperidin] -l'-yl carbonyl3-2- (phenylmethyl-oxy) ethyl] -2-amino-2-methylpropanamide alsulfonate (16) Materials: Amine (15) 2340 g (4.43 moles) Methanesulfonic Acid (MsOH) 316 mL (4.88 moles) EtOAc 60 L EtOH 4.8 L 8% EtOH in 20 L EtOAc The volume of the solution of 15 from the previous step was adjusted to 60 1 with ethyl acetate and EtOH (4.8 L) was added. MsOH (316 mL) was added in 3 1 of EtOAc at 45 ° C. To the homogeneous deep red solution 496 g of the seed of form I of the title compound were added (10% of seed was used based on the weight of the free amine) the temperature was raised to approximately 48 ° C and the reaction was aged at 52 ° C for 1.5 hours. The analysis indicated complete conversion to the title compound (form I). (Less than 10% of seed was required greater than the aging time (>;3 hours)). The suspension was allowed to cool to 20 ° C overnight and was filtered in a low centrifuge 2. The cake was washed with 20 L of 8% EtOH in EtOAc. N2 is essential during filtration because the wet crystals are very hydroscopic. The batch was dried at 35 ° C under vacuum to give 2.7 Kg (56% overall yield) of the title compound (form I) (purity 99.9 A%; <0.15 enantiomer). The conversion of form II to form I is also achieved where the salt is formed in EtOAc-EtOH by the addition of MsOH as before and the initial solution of the salt (at 55 ° C) is cooled to 45 ° C . The crystals start to appear at that temperature and the suspension becomes thicker with time. The temperature is then raised to 51 ° C and the suspension is aged overnight. Complete conversion to form I of 16 would be expected. This procedure can also be used to prepare seed crystals of Form I of 16.

EXAMPLE 15B Spiro methansulfonate [3H-indol-3,4'-piperidin] -l'-yl) cartilane3-2- (phenylmethyl-oxy) ethyl-3-amino-2-methylpropanamide I161 Materials: Amine (15) 3.1 Kg (5.86 moles) Methanesulfonic acid 620 g (6.45 moles) Ethyl acetate 37 L Absolute ethanol 8.7 L Spiro methansulfonate [3H-indol-3,4'-piperdin] - l'-il) - carbonyl] -2- (phenylmethyloxy) ethyl] -2-amino-2-methylpropane ida (Form I) 70 g (0.11 moles) Absolute ethanol (6.4 1) was added to the solution of the amine (15) (3.1 kg) in ethyl acetate (total volume ~62) in a reaction vessel. The batch was heated to 50 ° C and a solution of methanesulfonic acid (620 g, 412 ml, 1.1 equiv.) In ethyl acetate (11 L) was added for ~ 5 minutes at 50-54 ° C. The batch was seeded with spiro methanesulfonate [3 H -indole-3,4'-piperidin] -l'-1) carbonyl] -2- (phenylmethyloxy) ethyl 2-amino-2-methylpropanamide (Form I) (70 g ) and the resulting suspension was stirred and heated to 55 ° C under nitrogen atmosphere overnight. The next day, the suspension was cooled to 15-20 ° C, maintained for 2 hours and then dropped to 50 μm of polypropylene filter under a nitrogen atmosphere. The solid product was washed with a mixture of absolute ethanol (2.3 1) in ethyl acetate (26 L). The white solid product was excavated and dried in an Apex vacuum oven at 35 ° C for an appropriate time of about two days). Spiro methanesulfonate [3H-indol-3,4'-piperidin] -l '-yl) -carbonyl] -2- (phenylmethyl-oxy) and thi] -2-amino-2-methylpropanamide (3,352 kg) dry was screened using a Jackson Crockatt sieve to give 3347 kg (including seed, 70 g)} yield = 3,277 kg. Form I of N- [1 (R) - [(1,2-dihydro-l-methanesulfonyl-spiro [3H-indol-3,4'-piperin] -l'-yl) carbonyl] -2- sulfonate (phenylmethyl-oxy) ethyl] -2-amino-2-methylpropanamide is an anhydrous polymorph characterized by the following properties: a melting point of 168-171 ° C and solubility in isopropanol of 4.6 mg / mL. The DSC curve for the methanesulfonate form I of N-Cl (R) - [1,2-dihydro-l-methanesulfonyl-spiro [3H-indole-3,4'-pipe rdi n] -1 '-i 1) carbonyl] -2- (feni lmeti loxi) eti l] -2-amino-2-methylpropanamide at 10 ° C / min in an open cup under nitrogen flow shows an individual endotherm, due to melting, with a peak temperature of about 180 ° C and an extrapolated onset temperature (melting point) of about 170 ° C with an associated heat of about 53 J / g. Form I was characterized by an X-ray powder diffraction pattern with reflections at approximately: 6.5, 14.7, 16.9, 17.1, 17.9, 19.5, 21.1, 21.7 and 22.0 ° (2 teta). Data collected using an APD3720 automated powder diffraction instrument with copper Ka radiation. Measurements were made from 2o to 40 ° (2 teta) with the sample obtained at room temperature.

CLAR conditions: The retention times of liquid chromatography on Zorbax RX-C8 (4.6 mm x 25 cm), wavelength = 210 nm, flow rate = 1.5 ml / min. Compound 1: 60:40 CH3CN-H2O (1% H3PO4), RT = 5.0 min. Compound Ib: 35:65 CH3CN-H2O (0.1% NHAOAC), RT = 6.2 min. Compound 10: 60:40 CH3CN-H2O (0.1% H3PO4), RT = 2.9 min.

Compound 11: 60:40 CH3CN-H2O (0.1% H3PO4), RT = 5.4 min. Compound 12: 40:60 CH3CN-H2O [pH 5.25 NaH2 P0 ((6.9 g / L H2O) is adjusted with NaOH)], RT = 5.6 min. Compound 14: 60:40 CH3CN-H2O (0.1% H3PO4), RT = 4.65 min. Compound 15: 40:60 CH3CN-H2OCPH = 5.25 NaH2P0 «(6.9 g / L H2O adjusted with NaOH), TR = 4.9 min.

Retention time of liquid chromatography on Zorbax RX-C8 (4.6 mm x 25 cm), wavelength = 210 nm, flow rate = 1.2 ml / min, column temperature = 48 ° C. Solvent A = 0.05% phosphoric acid + 0.01% triethylamine in water. Solvent B = acetonitrile Gradient system: Time% A% B 0 min 95 5 35 min 10 90 38 min 95 5 40 min 95 5 Retention time (minutes) Compound 1 25.2 Compound Ib 8.5 Compound 10 20.5 Compound 11 26.3 Compound 12 14.8 Compound 14 25.6 Compound 15 15.7 Although the invention has been described and illustrated with reference to certain particular embodiments thereof, those skilled in the art will appreciate that various adaptations, changes, modifications, substitutions, deletions or additions of procedures and protocols can be made without departing from the spirit and scope. of the invention. For example, reaction conditions other than the particular conditions as set forth above may be applicable as a consequence of variations in the reagents or methodology for preparing the compounds of the methods of the invention indicated above.

Also, the specific reactivity of starting materials may vary according to the particular substituents present or the manufacturing conditions or depending on them, and said expected variations or differences in the results are contemplated in accordance with the objects and practices of the present invention. Therefore, it is intended that the invention be defined by the scope of the following claims and that said claims be interpreted as widely as is reasonable.

Claims (28)

NOVELTY OF THE INVENTION CLAIMS

1. - A process for the preparation of a compound of the formula V: comprising: (1) copying an amino acid of the formula:

^ C02H Ph 'Ñ-L H with a compound of the formula: H

Me in the presence of a first acid activating agent in a first inert solvent in the presence of the first catalytic agent to give a compound of the formula I: wherein L is an amino protecting group followed by: (2) reacting the compound of the formula I with a first amino deprotecting agent to give a compound of the formula II: followed by: (3) copulate an amino acid of the formula: wherein L is an amino protecting group, with the compound of formula II in the presence of a second acid activating agent in a solvent comprising acetonitronil or isopropyl acetate: water in the presence of a second catalytic agent, to give a compound of formula III:

III wherein L is an amino protecting group, followed by: (4) reacting the compound of formula III with a second amino deprotecting agent to give a compound of formula IV, or a pharmaceutically acceptable salt thereof:

IV followed by reacting the compound of the formula IV with a methanesulfonic acid to give the compound of the formula V. 2. A process for the preparation of a compound of the formula I: wherein L is an amino protecting group, coupling an amino acid of the formula:

N-L H with a compound of the formula: in the presence of an acid activating agent in an inert solvent in the presence of a catalytic agent to give the compound of formula I. 3. The process according to claim 2, further characterized in that the solvent comprises isopropyl acetate: water . 4. The process according to claim 2, further characterized in that the acid activating agent is DCC. 5. The process according to claim 2, further characterized in that the catalytic agent is HOBT. 6. The process according to claim 2, further characterized in that the solvent further comprises another solvent that is selected from the group consisting of: ethyl acetate, propionitrile; chlorinated hydrocarbon selected from chloromethane, chloroform, carbon tetrachloride, dichloroethane, dichlorobenzene, and orthodichlorobenzene; benzene; toluene; xylenes and mixtures thereof.

7. The process according to claim 2, further characterized in that the temperature of the reaction is between 20 and 35 ° C.

8. The process according to claim 2, further characterized in that the compound of the formula I, in the amino protecting group is selected from: t-butoxycarbonyl.

9. The process according to claim 2, which is conducted in situ without isolation of the compound of the formula I following its preparation.

10. A process for the preparation of the compound of the formula II of claim 1: II which comprises reacting a compound of the formula I wherein L is an amino protecting group, with an amino deprotecting agent to give the compound of formula II.

11. The process according to claim 10, further characterized in that in the compound of the formula I, the amino protecting group is selected from: t-butoxycarbonyl.

12. The process according to claim 10, further characterized in that the amino deprotective agent is methanesulfonic acid.

13. The process according to claim 10, which is conducted in situ without isolation of the compound of the formula II, after its preparation.

14. A process for the preparation of a compound of the formula III: III wherein L is an amino protecting group, coupling an amino acid of the formula: wherein L is an amino protecting group, with a compound of formula II: in the presence of an acid activating agent in a solvent comprising acetonitrile or isopropyl acetate: water in the presence of a catalyst agent, to give the compound of formula III.

15. The process according to claim 14, further characterized in that the solvent comprises isopropyl acetate: water.

16. The process according to claim 15, further characterized in that the solvent isopropyl acetate: water is in a ratio of about 40:60 to 60:40 (by volume).

17. The process according to claim 14, further characterized in that the acid activating agent is DCC.

18. The process according to claim 14, further characterized in that the catalytic agent is HOBT.

19. - The method according to claim 14, further characterized in that the temperature of the reaction is between 20 and 35 ° C.

20. The process according to claim 14, further characterized in that the compound of the formula III, the amino protecting group is selected from: t-butoxycarbonyl.

21. The process according to claim 14, which is conducted in situ without isolation of the compound of the formula III following its preparation.

22. A process for the preparation of the compound of formula IV of claim 1 or a pharmaceutically acceptable salt thereof: IV comprising reacting a compound of formula III wherein L is an amino protecting group, with an amino deprotecting agent to give the compound of the formula IV.

23. The process according to claim 22, further characterized in that in the compound of the formula III, the The amino protecting group is selected from: t-butoxycarbonyl.

24. The process according to claim 22, further characterized in that the amino deprotecting agent is methanesulfonic acid.

25. The process according to claim 22 which is conducted in a solution comprising ethanol.

26. The process according to claim 22, which is conducted in situ without isolation of the compound of the formula IV after its preparation.

27. A process for the preparation of the compound of formula V of claim 1: SO2Me which comprises makes a compound of the formula V react: with methanesulfonic acid to give the compound of formula V. 28.- The process according to claim 27 which is conducted in a solution comprising ethyl acetate and ethanol.