SMALL MOLECULES FOR BORON NEUTRON CAPTURE THERAPY

Abstract

Disclosed are compounds, compositions, and methods useful for boron neutron capture therapy.

Claims

1. A compound of Formula (I): ##STR00030## wherein Y.sub.1 is absent or O; Y.sub.2 is optionally substituted alkylene; R.sub.1, R.sub.2, and Rs are each independently selected from H and halo; R.sub.3 is selected from H, halo, and X.sub.1-X.sub.2; R.sub.4 is selected from H, halo, and X.sub.1-X.sub.2; X.sub.1 is alkylene; X.sub.2 is C(H)(NH.sub.2)CO.sub.2H; and the compound is racemic, enriched in one enantiomer, or a single enantiomer; provided that the compound comprises one and only one occurrence of X.sub.1-X.sub.2; or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1 having the structure of Formula (IA): ##STR00031## wherein R.sub.4 is selected from H and halo.

3. The compound of claim 2, wherein each of R.sub.1, R.sub.2, R.sub.4, and R.sub.5 is H; or one of R.sub.1, R.sub.2, R.sub.4, and R.sub.5 is halo, and each of the remainder of R.sub.1, R.sub.2, R.sub.4, and R.sub.5 is H.

4. (canceled)

5. (canceled)

6. The compound of claim 2 having the structure selected from: ##STR00032## wherein * indicates a chiral carbon with an absolute configuration of (S) or (R); and the compound is not racemic.

7. The compound of claim 6, wherein the absolute configuration of the chiral carbon is (S).

8. The compound of claim 1 having the structure of Formula (TB): ##STR00033## wherein R.sub.3 is selected from H and halo.

9. The compound of claim 8, wherein each of R.sub.1, R.sub.2, R.sub.3, and R.sub.5 is H; or one of R.sub.1, R.sub.2, R.sub.4, and R.sub.5 is halo, and each of the remainder of R.sub.1, R.sub.2, R.sub.4, and R.sub.5 is H.

10. (canceled)

11. (canceled)

12. The compound of claim 8 having the structure selected from: ##STR00034## wherein * indicates a chiral carbon with an absolute configuration of (S) or (R); and the compound is not racemic.

13. The compound of claim 12, wherein the absolute configuration of the chiral carbon is (S).

14. The compound of claim 1, wherein X.sub.1 is (C.sub.1-C.sub.4)alkylene.

15. (canceled)

16. The compound of claim 1, wherein Y.sub.1 is O.

17. The compound of claim 1, wherein Y.sub.1 is absent.

18. The compound of claim 16, wherein Y.sub.2 is unsubstituted (C.sub.1-C.sub.4)alkylene.

19. (canceled)

20. (canceled)

21. The compound of claim [[20]]16, wherein Y.sub.2 is seleted from C(Y.sub.3)(Y.sub.4) and C(Y.sub.3)(Y.sub.4)CH.sub.2; and Y.sub.3 and Y.sub.4 are each independently selected from H, halo, alkyl, and heteroalkyl, provided that at least one of Y.sub.3 and Y.sub.4 is not H; or Y.sub.3 and Y.sub.4 taken together with the carbon to which they are bonded form a cycloalkyl, cycloheteroalkyl, spiro cycloalkyl or spiro cycloheteroalkyl.

22. (canceled)

23. The compound of claim 17, wherein Y.sub.2 is unsubstituted (C.sub.1-C.sub.4)alkylene.

24. (canceled)

25. (canceled)

26. The compound of claim 17, wherein Y.sub.2 is seleted from C(Y.sub.3)(Y.sub.4) and C(Y.sub.3)(Y.sub.4)CH.sub.2; and Y.sub.3 and Y.sub.4 are each independently selected from H, halo, alkyl, and heteroalkyl, provided that at least one of Y.sub.3 and Y.sub.4 is not H; or Y.sub.3 and Y.sub.4 taken together with the carbon to which they are bonded form a cycloalkyl, cycloheteroalkyl, spiro cycloalkyl or spiro cycloheteroalkyl.

27. (canceled)

28. The compound of claim 1 having the structure selected from: ##STR00035## ##STR00036## ##STR00037## or a pharmaceutically acceptable salt thereof.

29. (canceled)

30. The compound of claim 1, wherein the boron atom in the compound is .sup.10B.

31. A pharmaceutical composition, comprising a compound of claim 1; and a pharmaceutical acceptable excipient.

32. (canceled)

33. (canceled)

34. (canceled)

35. A method of treating cancer, comprising: i) administering to a subject in need thereof a compound of claim 1, wherein the compound accumulates in a plurality of cancer cells in the subject; and ii) irradiating the plurality of cancer cells with neutrons.

36.-46. (canceled)

47. The method of claim 35, wherein the cancer is selected from head and neck cancer, glioblastoma, melanoma, sarcoma, breast cancer, meningioma, lung cancer, mesothelioma, hepatocellular carcinoma, and extramammary Paget disease.

48. (canceled)

Description

DETAILED DESCRIPTION

Definitions

[0020] For convenience, before further description of the present invention, certain terms employed in the specification, examples and appended claims are collected here. These definitions should be read in light of the remainder of the disclosure and understood as by a person of skill in the art. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by a person of ordinary skill in the art.

[0021] In order for the present invention to be more readily understood, certain terms and phrases are defined below and throughout the specification.

[0022] The articles a and an are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, an element means one element or more than one element.

[0023] The phrase and/or, as used herein in the specification and in the claims, should be understood to mean either or both of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Multiple elements listed with and/or should be construed in the same fashion, i.e., one or more of the elements so conjoined. Other elements may optionally be present other than the elements specifically identified by the and/or clause, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, a reference to A and/or B, when used in conjunction with open-ended language such as comprising can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc.

[0024] As used herein in the specification and in the claims, or should be understood to have the same meaning as and/or as defined above. For example, when separating items in a list, or or and/or shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly indicated to the contrary, such as only one of or exactly one of, or, when used in the claims, consisting of, will refer to the inclusion of exactly one element of a number or list of elements. In general, the term or as used herein shall only be interpreted as indicating exclusive alternatives (i.e., one or the other but not both) when preceded by terms of exclusivity, such as either, one of, only one of, or exactly one of. Consisting essentially of, when used in the claims, shall have its ordinary meaning as used in the field of patent law.

[0025] As used herein in the specification and in the claims, the phrase at least one, in reference to a list of one or more elements, should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. This definition also allows that elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase at least one refers, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, at least one of A and B (or, equivalently, at least one of A or B, or, equivalently at least one of A and/or B) can refer, in one embodiment, to at least one, optionally including more than one, A, with no B present (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements); etc.

[0026] It should also be understood that, unless clearly indicated to the contrary, in any methods claimed herein that include more than one step or act, the order of the steps or acts of the method is not necessarily limited to the order in which the steps or acts of the method are recited.

[0027] In the claims, as well as in the specification above, all transitional phrases such as comprising, including, carrying, having, containing, involving, holding, composed of, and the like are to be understood to be open-ended, i.e., to mean including but not limited to. Only the transitional phrases consisting of and consisting essentially of shall be closed or semi-closed transitional phrases, respectively, as set forth in the United States Patent Office Manual of Patent Examining Procedures, Section 2111.03.

[0028] Certain compounds contained in compositions of the present invention may exist in particular geometric or stereoisomeric forms. In addition, polymers of the present invention may also be optically active. The present invention contemplates all such compounds, including cis-and trans-isomers, R- and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the invention. Additional asymmetric carbon atoms may be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in this invention.

[0029] Geometric isomer means isomers that differ in the orientation of substituent atoms in relationship to a carbon-carbon double bond, to a cycloalkyl ring, or to a bridged bicyclic system. Atoms (other than H) on each side of a carbon- carbon double bond may be in an E (substituents are on opposite sides of the carbon- carbon double bond) or Z (substituents are oriented on the same side) configuration. R, S, S*, R*, E, Z, cis, and trans, indicate configurations relative to the core molecule. Certain of the disclosed compounds may exist in atropisomeric forms or as atropisomers. Atropisomers are stereoisomers resulting from hindered rotation about single bonds where the steric strain barrier to rotation is high enough to allow for the isolation of the conformers. The compounds of the invention may be prepared as individual isomers by either isomer-specific synthesis or resolved from a mixture of isomers. Conventional resolution techniques include forming the salt of a free base of each isomer of an isomeric pair using an optically active acid (followed by fractional crystallization and regeneration of the free base), forming the salt of the acid form of each isomer of an isomeric pair using an optically active amine (followed by fractional crystallization and regeneration of the free acid), forming an ester or amide of each of the isomers of an isomeric pair using an optically pure acid, amine or alcohol (followed by chromatographic separation and removal of the chiral auxiliary), or resolving an isomeric mixture of either a starting material or a final product using various well known chromatographic methods.

[0030] If, for instance, a particular enantiomer of compound of the present invention is desired, it may be prepared by asymmetric synthesis, or by derivation with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers. Alternatively, where the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts are formed with an appropriate optically-active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographic means well known in the art, and subsequent recovery of the pure enantiomers.

[0031] Percent purity by mole fraction is the ratio of the moles of the enantiomer (or diastereomer) or over the moles of the enantiomer (or diastereomer) plus the moles of its optical isomer. When the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least about 60%, about 70%, about 80%, about 90%, about 99% or about 99.9% by mole fraction pure relative to the other stereoisomers. When a single enantiomer is named or depicted by structure, the depicted or named enantiomer is at least about 60%, about 70%, about 80%, about 90%, about 99% or about 99.9% by mole fraction pure. When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is at least about 60%, about 70%, about 80%, about 90%, about 99% or about 99.9% by mole fraction pure.

[0032] When a disclosed compound is named or depicted by structure without indicating the stereochemistry, and the compound has at least one chiral center, it is to be understood that the name or structure encompasses either enantiomer of the compound free from the corresponding optical isomer, a racemic mixture of the compound or mixtures enriched in one enantiomer relative to its corresponding optical isomer. When a disclosed compound is named or depicted by structure without indicating the stereochemistry and has two or more chiral centers, it is to be understood that the name or structure encompasses a diastereomer free of other diastereomers, a number of diastereomers free from other diastereomeric pairs, mixtures of diastereomers, mixtures of diastereomeric pairs, mixtures of diastereomers in which one diastereomer is enriched relative to the other diastereomer(s) or mixtures of diastereomers in which one or more diastereomer is enriched relative to the other diastereomers. The invention embraces all of these forms.

[0033] Structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds produced by the replacement of a hydrogen with deuterium or tritium, or of a carbon with a .sup.13C- or .sup.14C-enriched carbon, or of a boron with .sup.10B-enriched boron, are within the scope of this invention.

[0034] The term prodrug as used herein encompasses compounds that, under physiological conditions, are converted into therapeutically active agents. A common method for making a prodrug is to include selected moieties that are hydrolyzed under physiological conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by an enzymatic activity of the host animal.

[0035] The phrase pharmaceutically acceptable excipient or pharmaceutically acceptable carrier as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject chemical from one organ or portion of the body, to another organ or portion of the body. Each carrier must be acceptable in the sense of being compatible with the other ingredients of the formulation, not injurious to the patient, and substantially non-pyrogenic. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations. In certain embodiments, pharmaceutical compositions of the present invention are non-pyrogenic, i.e., do not induce significant temperature elevations when administered to a patient.

[0036] The term pharmaceutically acceptable salts refers to the relatively non-toxic, inorganic and organic acid addition salts of the compound(s). These salts can be prepared in situ during the final isolation and purification of the compound(s), or by separately reacting a purified compound(s) in its free base form with a suitable organic or inorganic acid, and isolating the salt thus formed. Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, and laurylsulphonate salts, and the like. (See, for example, Berge et al. (1977) Pharmaceutical Salts, J. Pharm. Sci. 66:1-19.)

[0037] In other cases, the compounds useful in the methods of the present invention may contain one or more acidic functional groups and, thus, are capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable bases. The term pharmaceutically acceptable salts in these instances refers to the relatively non-toxic inorganic and organic base addition salts of a compound(s). These salts can likewise be prepared in situ during the final isolation and purification of the compound(s), or by separately reacting the purified compound(s) in its free acid form with a suitable base, such as the hydroxide, carbonate, or bicarbonate of a pharmaceutically acceptable metal cation, with ammonia, or with a pharmaceutically acceptable organic primary, secondary, or tertiary amine. Representative alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, and aluminum salts, and the like. Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like (see, for example, Berge et al., supra).

[0038] The term pharmaceutically acceptable cocrystals refers to solid coformers that do not form formal ionic interactions with the small molecule.

[0039] A therapeutically effective amount (or effective amount) of a compound with respect to use in treatment, refers to an amount of the compound in a preparation which, when administered as part of a desired dosage regimen (to a mammal, preferably a human) alleviates a symptom, ameliorates a condition, or slows the onset of disease conditions according to clinically acceptable standards for the disorder or condition to be treated or the cosmetic purpose, e.g., at a reasonable benefit/risk ratio applicable to any medical treatment.

[0040] The term prophylactic or therapeutic treatment is art-recognized and includes administration to the host of one or more of the subject compositions. If it is administered prior to clinical manifestation of the unwanted condition (e.g., disease or other unwanted state of the host animal) then the treatment is prophylactic, (i.e., it protects the host against developing the unwanted condition), whereas if it is administered after manifestation of the unwanted condition, the treatment is therapeutic, (i.e., it is intended to diminish, ameliorate, or stabilize the existing unwanted condition or side effects thereof).

[0041] The term patient or subject refers to a mammal in need of a particular treatment. In certain embodiments, a patient is a primate, canine, feline, or equine. In certain embodiments, a patient is a human.

[0042] An aliphatic chain comprises the classes of alkyl, alkenyl and alkynyl defined below. A straight aliphatic chain is limited to unbranched carbon chain moieties. As used herein, the term aliphatic group refers to a straight chain, branched-chain, or cyclic aliphatic hydrocarbon group and includes saturated and unsaturated aliphatic groups, such as an alkyl group, an alkenyl group, or an alkynyl group.

[0043] Alkyl refers to a fully saturated cyclic or acyclic, branched or unbranched carbon chain moiety having the number of carbon atoms specified, or up to 30 carbon atoms if no specification is made. For example, alkyl of 1 to 8 carbon atoms refers to moieties such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, and octyl, and those moieties which are positional isomers of these moieties. Alkyl of 10 to 30 carbon atoms includes decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, heneicosyl, docosyl, tricosyl and tetracosyl. In certain embodiments, a straight chain or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., C1-C30 for straight chains, C3-C30for branched chains), and more preferably 20 or fewer. Alkyl goups may be substituted or unsubstituted.

[0044] As used herein, the term heteroalkyl refers to an alkyl moiety as hereinbefore defined which contain one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms in place of carbon atoms.

[0045] As used herein, the term haloalkyl refers to an alkyl group as hereinbefore defined substituted with at least one halogen.

[0046] As used herein, the term hydroxyalkyl refers to an alkyl group as hereinbefore defined substituted with at least one hydroxyl.

[0047] As used herein, the term alkylene refers to an alkyl group having the specified number of carbons, for example from 1 to 12 carbon atoms, that contains two points of attachment to the rest of the compound on its longest carbon chain. Non-limiting examples of alkylene groups include methylene (CH.sub.2), ethylene (CH.sub.2CH.sub.2), n-propylene (CH.sub.2CH.sub.2CH.sub.2), isopropylene (CH.sub.2CH(CH.sub.3)), and the like. Alkylene groups can be cyclic or acyclic, branched or unbranched carbon chain moiety, and may be optionally substituted with one or more substituents.

[0048] Cycloalkyl means mono- or bicyclic or bridged or spirocyclic, or polycyclic saturated carbocyclic rings, each having from 3 to 12 carbon atoms. Preferred cycloalkyls have from 3-10 carbon atoms in their ring structure, and more preferably have 3-6 carbons in the ring structure. Cycloalkyl groups may be substituted or unsubstituted.

[0049] As used herein, the term halocycloalkyl refers to a cycloalkyl group as hereinbefore defined substituted with at least one halogen.

[0050] Cycloheteroalkyl refers to a cycloalkyl moiety as hereinbefore defined which contain one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms in place of carbon atoms. Preferred cycloheteroalkyls have from 4-8 carbon atoms and heteroatoms in their ring structure, and more preferably have 4-6 carbons and heteroatoms in the ring structure. Cycloheteroalkyl groups may be substituted or unsubstituted.

[0051] Unless the number of carbons is otherwise specified, lower alkyl, as used herein, means an alkyl group, as defined above, but having from one to ten carbons, more preferably from one to six carbon atoms in its backbone structure such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. Likewise, lower alkenyl and lower alkynyl have similar chain lengths. Throughout the application, preferred alkyl groups are lower alkyls. In certain embodiments, a substituent designated herein as alkyl is a lower alkyl.

[0052] Alkenyl refers to any cyclic or acyclic, branched or unbranched unsaturated carbon chain moiety having the number of carbon atoms specified, or up to 26 carbon atoms if no limitation on the number of carbon atoms is specified; and having one or more double bonds in the moiety. Alkenyl of 6 to 26 carbon atoms is exemplified by hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, nonadecenyl, eicosenyl, heneicosoenyl, docosenyl, tricosenyl, and tetracosenyl, in their various isomeric forms, where the unsaturated bond(s) can be located anywhere in the moiety and can have either the (Z) or the (E) configuration about the double bond(s).

[0053] Alkynyl refers to hydrocarbyl moieties of the scope of alkenyl, but having one or more triple bonds in the moiety.

[0054] The term aryl as used herein includes 3- to 12-membered substituted or unsubstituted single-ring aromatic groups in which each atom of the ring is carbon (i.e., carbocyclic aryl) or where one or more atoms are heteroatoms (i.e., heteroaryl). Preferably, aryl groups include 5- to 12-membered rings, more preferably 6- to 10-membered rings The term aryl also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is aromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Carboycyclic aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like. Heteroaryl groups include substituted or unsubstituted aromatic 3- to 12-membered ring structures, more preferably 5- to 12-membered rings, more preferably 5- to 10-membered rings, whose ring structures include one to four heteroatoms. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like. Aryl and heteroaryl can be monocyclic, bicyclic, or polycyclic.

[0055] The term halo, halide, or halogen as used herein means halogen and includes, for example, and without being limited thereto, fluoro, chloro, bromo, iodo and the like, in both radioactive and non-radioactive forms. In a preferred embodiment, halo is selected from the group consisting of fluoro, chloro and bromo.

[0056] The terms heterocyclyl or heterocyclic group refer to 3- to 12-membered ring structures, more preferably 5- to 12-membered rings, more preferably 5- to 10-membered rings, whose ring structures include one to four heteroatoms. Heterocycles can be monocyclic, bicyclic, spirocyclic, or polycyclic. Heterocyclyl groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, xanthene, phenoxathiin, pyrrole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, phenarsazine, phenothiazine, furazan, phenoxazine, pyrrolidine, oxolane, thiolane, oxazole, piperidine, piperazine, morpholine, lactones, lactams such as azetidinones and pyrrolidinones, sultams, sultones, and the like. The heterocyclic ring can be substituted at one or more positions with such substituents as described above, as for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphate, phosphonate, phosphinate, carbonyl, carboxyl, silyl, sulfamoyl, sulfinyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, a heterocyclyl, an aromatic or heteroaromatic moiety, CF.sub.3, CN, and the like.

[0057] The term substituted refers to moieties having substituents replacing a hydrogen on one or more carbons of the backbone. It will be understood that substitution or substituted with includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. As used herein, the term substituted is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. Substituents can include any substituents described herein, for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an alkoxy, a phosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety. In preferred embodiments, the substituents on substituted alkyls are selected from C1_6 alkyl, C3-6 cycloalkyl, halogen, carbonyl, cyano, or hydroxyl. In more preferred embodiments, the substituents on substituted alkyls are selected from fluoro, carbonyl, cyano, or hydroxyl. It will be understood by those skilled in the art that substituents can themselves be substituted, if appropriate. Unless specifically stated as unsubstituted, references to chemical moieties herein are understood to include substituted variants. For example, reference to an aryl group or moiety implicitly includes both substituted and unsubstituted variants.

[0058] As used herein, the definition of each expression, e.g., alkyl, m, n, etc., when it occurs more than once in any structure, is intended to be independent of its definition elsewhere in the same structure.

[0059] As used herein, small molecules refers to small organic or inorganic molecules of molecular weight below about 3,000 Daltons. In general, small molecules useful for the invention have a molecular weight of less than 3,000 Daltons (Da). The small molecules can be, e.g., from at least about 100 Da to about 3,000 Da (e.g., between about 100 to about 3,000 Da, about 100 to about 2500 Da, about 100 to about 2,000 Da, about 100 to about 1,750 Da, about 100 to about 1,500 Da, about 100 to about 1,250 Da, about 100 to about 1,000 Da, about 100 to about 750 Da, about 100 to about 500 Da, about 200 to about 1500, about 500 to about 1000, about 300 to about 1000 Da, or about 100 to about 250 Da).

[0060] In some embodiments, a small molecule refers to an organic, inorganic, or organometallic compound typically having a molecular weight of less than about 1000. In some embodiments, a small molecule is an organic compound, with a size on the order of 1 nm. In some embodiments, small molecule drugs of the invention encompass oligopeptides and other biomolecules having a molecular weight of less than about 1000.

[0061] An effective amount is an amount sufficient to effect beneficial or desired results. For example, a therapeutic amount is one that achieves the desired therapeutic effect. This amount can be the same or different from a prophylactically effective amount, which is an amount necessary to prevent onset of disease or disease symptoms. An effective amount can be administered in one or more administrations, applications or dosages. A therapeutically effective amount of a composition depends on the composition selected. The compositions can be administered from one or more times per day to one or more times per week; including once every other day. The skilled artisan will appreciate that certain factors may influence the dosage and timing required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the general health and/or age of the subject, and other diseases present. Moreover, treatment of a subject with a therapeutically effective amount of the compositions described herein can include a single treatment or a series of treatments. A series of treatments may comprise a second or subsequent treatment weeks to months after a first or preceding treatment.

[0062] The terms decrease, reduce, reduced, reduction, decrease, and inhibit are all used herein generally to mean a decrease by a statistically significant amount relative to a reference. However, for avoidance of doubt, reduce, reduction or decrease or inhibit typically means a decrease by at least 10% as compared to a reference level and can include, for example, a decrease by at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, up to and including, for example, the complete absence of the given entity or parameter ascompared to the reference level, or any decrease between 10-99% as compared to the absence of a given treatment.

[0063] The terms increased, increase or enhance or activate are all used herein to generally mean an increase by a statically significant amount; for the avoidance of any doubt, the terms increased, increase or enhance or activate means an increase of at least 10% as compared to a reference level, for example an increase of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% increase or any increase between 10-100% as compared to a reference level, or at least about a 2-fold, or at least about a 3-fold, or at least about a 4-fold, or at least about a 5-fold or at least about a 10-fold increase, or any increase between 2-fold and 10-fold or greater as compared to a reference level.

[0064] As used herein, the term modulate includes up-regulation and down-regulation, e.g., enhancing or inhibiting a response.

[0065] A radiopharmaceutical agent, as defined herein, refers to a pharmaceutical agent which contains at least one radiation-emitting radioisotope. Radiopharmaceutical agents are routinely used in nuclear medicine for the diagnosis and/or therapy of various diseases. The radiolabelled pharmaceutical agent, for example, a radiolabelled antibody, contains a radioisotope (RI) which serves as the radiation source. As contemplated herein, the term radioisotope includes metallic and non-metallic radioisotopes. The radioisotope is chosen based on the medical application of the radiolabeled pharmaceutical agents. When the radioisotope is a metallic radioisotope, a chelator is typically employed to bind the metallic radioisotope to the rest of the molecule. When the radioisotope is a non-metallic radioisotope, the non-metallic radioisotope is typically linked directly, or via a linker, to the rest of the molecule.

[0066] For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover.

Compounds of the Invention

[0067] One aspect of the invention relates to a compound of Formula (I):

##STR00002## [0068] wherein [0069] Y.sub.1 is absent or O; [0070] Y.sub.2 is optionally substituted alkylene; [0071] R.sub.1, R.sub.2, and R.sub.5 are each independently selected from H and halo; [0072] R.sub.3 is selected from H, halo, and X.sub.1-X.sub.2; [0073] R.sub.4 is selected from H, halo, and X.sub.1-X.sub.2; [0074] X.sub.1 is alkylene; [0075] X.sub.2 is C(H)(NH.sub.2)CO.sub.2H; and [0076] the compound is racemic, enriched in one enantiomer, or a single enantiomer; [0077] provided that the compound comprises one and only one occurrence of X.sub.1-X.sub.2; or a pharmaceutically acceptable salt thereof.

[0078] In certain embodiments, the compound having the structure of Formula (IA):

##STR00003## [0079] wherein R.sub.4 is selected from H and halo.

[0080] In certain embodiments, each of R.sub.1, R.sub.2, R.sub.4, and R.sub.5 is H.

[0081] In certain embodiments, one of R.sub.1, R.sub.2, R.sub.4, and R.sub.5 is halo, and each of the remainder of R.sub.1, R.sub.2, R.sub.4, and R.sub.5 is H.

[0082] In certain embodiments, the halo is F.

[0083] In certain embodiments, the compound having the structure selected from:

##STR00004##

wherein * indicates a chiral carbon with an absolute configuration of (S) or (R); and the compound is not racemic.

[0084] In certain embodiments, the absolute configuration of the chiral carbon is (S).

[0085] In certain embodiments, the absolute configuration of the chiral carbon is (R).

[0086] In certain embodiments, the compound having the structure of Formula (TB):

##STR00005## [0087] wherein R.sub.3 is selected from H and halo.

[0088] In certain embodiments, each of R.sub.1, R.sub.2, R.sub.3, and R.sub.5 is H.

[0089] In certain embodiments, one of R.sub.1, R.sub.2, R.sub.3, and R.sub.5 is halo, and each of the remainder of R.sub.1, R.sub.2, R.sub.3, and R.sub.5 is hydrogen.

[0090] In certain embodiments, the halo is F.

[0091] In certain embodiments, the compound having the structure selected from:

##STR00006##

wherein * indicates a chiral carbon with an absolute configuration of (S) or (R); and the compound is not racemic.

[0092] In certain embodiments, the absolute configuration of the chiral carbon is (S).

[0093] In certain embodiments, the absolute configuration of the chiral carbon is (R).

[0094] In certain embodiments, X.sub.1 is (C.sub.1-C.sub.4)alkylene.

[0095] In certain embodiments, X.sub.1 is CH.sub.2.

[0096] In certain embodiments, Y.sub.1 is O.

[0097] In certain embodiments, Y.sub.2 is unsubstituted (C.sub.1C.sub.4)alkylene.

[0098] In certain embodiments, Y.sub.2 is selected from CH.sub.2 and CH.sub.2CH.sub.2.

[0099] In certain embodiments, Y.sub.2 is substituted (C.sub.1C.sub.4)alkylene.

[0100] In certain embodiments, Y.sub.2 is substituted with a halo, alkyl, heteroalkyl, cycloalkyl, or cyclohetero alkyl.

[0101] In certain embodiments, Y.sub.2 is selected from C(Y.sub.3)(Y.sub.4) and C(Y.sub.3)(Y.sub.4)CH.sub.2; and Y.sub.3 and Y.sub.4 are each independently selected from H, halo, alkyl, and heteroalkyl, provided that at least one of Y.sub.3 and Y.sub.4 is not H; or Y.sub.3 and Y.sub.4 taken together with the carbon to which they are bonded form a cycloalkyl, cycloheteroalkyl, spiro cycloalkyl or a spiro cycloheteroalkyl.

[0102] In certain embodiments, Y.sub.3 and Y.sub.4 taken together with the carbon to which they are bonded form a cyclopropyl.

[0103] In certain embodiments, Y.sub.1 is absent.

[0104] In certain embodiments, Y.sub.2 is unsubstituted (C.sub.1-C.sub.4)alkylene.

[0105] In certain embodiments, Y.sub.2 is selected from CH.sub.2 and CH.sub.2CH.sub.2.

[0106] In certain embodiments, Y.sub.2 is substituted (C.sub.1-C.sub.4)alkylene.

[0107] In certain embodiments, Y.sub.2 is substituted with a halo, alkyl, heteroalkyl, cycloalkyl, or cycloheteroalkyl.

[0108] In certain embodiments, Y.sub.2 is selected from C(Y.sub.3)(Y.sub.4) and C(Y.sub.3)(Y.sub.4)CH.sub.2; and Y.sub.3 and Y.sub.4 are each independently selected from H, halo, alkyl, and heteroalkyl, provided that at least one of Y.sub.3 and Y.sub.4 is not H; or Y.sub.3 and Y.sub.4 taken together with the carbon to which they are bonded form a cycloalkyl, cycloheteroalkyl, spiro cycloalkyl or spiro cycloheteroalkyl.

[0109] In certain embodiments, Y.sub.3 and Y.sub.4 taken together with the carbon to which they are bonded form a cyclopropyl.

[0110] In certain embodiments, the compound having the structure selected from:

##STR00007##

[0111] In certain embodiments, the compound having the structure selected from:

##STR00008##

[0112] In certain embodiments, the compound having the structure selected from:

##STR00009## ##STR00010## ##STR00011##

or a pharmaceutically acceptable salt thereof.

[0113] In certain embodiments, the compound having the structure:

##STR00012##

or a pharmaceutically acceptable salt thereof.

[0114] In certain embodiments, the boron atom in the compound is .sup.10B.

[0115] In certain embodiments, the compounds are atropisomers. Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds produced by the replacement of a hydrogen with deuterium or tritium, or of a carbon with a .sup.13C- or .sup.14C-enriched carbon are within the scope of this invention. Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents in accordance with the present invention. For example, in the case of variable R.sup.1, the (C.sub.1-C.sub.4)alkyl or the O(C.sub.1-C.sub.4)alkyl can be suitably deuterated (e.g., CD.sub.3, OCD.sub.3).

[0116] Also within the scope of the invention are compounds produced comprising the natural distribution of .sup.11B and .sup.10B.

[0117] Also within the scope of the invention are compounds enriched in .sup.10B, i.e., wherein the is .sup.10B present in abundance of >20%.

[0118] Any compound of the invention can also be radiolabed for the preparation of a radiopharmaceutical agent.

Methods of Treatment

[0119] One aspect of the invention provides compounds, compositions, and methods useful for boron neutron capture therapy.

[0120] Another aspect of the invention relates to a method of treating cancer, comprising: [0121] i) administering to a subject in need thereof a compound of Formula (I) or a pharmaceutical composition comprising a compound of Formula (I), wherein the compound accumulates in a plurality of cancer cells in the subject; and [0122] ii) irradiating the plurality of cancer cells with neutrons.

[0123] In certain embodiments, the compound selectively or preferentially accumulates in the plurality of cancer cells relative to noncancerous cells in the subject.

[0124] In certain embodiments, the irradiation results in conversion of a .sup.10B atom in the compound to an -particle and a lithium-7 ion.

[0125] In certain embodiments, the compound or the composition is administered intravenously.

[0126] In certain embodiments, the compound is continually administered during irradiation with neutrons.

[0127] In certain embodiments, in step (i) the compound is administered at about 100 mg/kg/h to about 500 mg/kg/h for a first period of time.

[0128] In certain embodiments, in step (i) the compound is administered at about 150 mg/kg/h to about 300 mg/kg/h for a first period of time.

[0129] In certain embodiments, the first period of time is about 1 hour to about 3 hours. In certain embodiments, the first period of time is about 2 hours.

[0130] In certain embodiments, in step (ii) the compound is administered at about 50 mg/kg/h to about 150 mg/kg/h for a second period of time.

[0131] In certain embodiments, in step (ii) the compound is administered at about 100 mg/kg/h to about 200 mg/kg/h for a second period of time.

[0132] In certain embodiments, the second period of time is about 0.25 hour to about 1.25 hours.

[0133] In certain embodiments, the second period of time is about 0.5 to about 1 hours.

[0134] In certain embodiments, the cancer is a solid tumor.

[0135] In certain embodiments, the cancer is selected from head and neck cancer, glioblastoma, melanoma, sarcoma, breast cancer, meningioma, lung cancer, mesothelioma, hepatocellular carcinoma, extramammary Paget disease.

[0136] In certain embodiments, the cancer is unresectable head and neck cancer.

[0137] In certain embodiments of any one of the disclosed methods, the compound of Formula (I) is defined as:

##STR00013## [0138] wherein [0139] Y.sub.1 is absent or O; [0140] Y.sub.2 is optionally substituted alkylene; [0141] R.sub.1, R.sub.2, and R.sub.5 are each independently selected from H and halo; [0142] R.sub.3 is selected from H, halo, and X.sub.1-X.sub.2; [0143] R.sub.4 is selected from H, halo, and X.sub.1-X.sub.2; [0144] X.sub.1 is alkylene; [0145] X.sub.2 is C(H)(NH.sub.2)CO.sub.2H; and [0146] the compound is racemic, enriched in one enantiomer, or a single enantiomer; [0147] provided that the compound comprises one and only one occurrence of X.sub.1-X.sub.2; [0148] or a pharmaceutically acceptable salt thereof.

Pharmaceutical Compositions, Routes of Administration, and Dosing

[0149] In certain embodiments, the invention is directed to a pharmaceutical composition, comprising a compound of the invention and a pharmaceutically acceptable carrier. In certain embodiments, the pharmaceutical composition comprises a plurality of compounds of the invention and a pharmaceutically acceptable carrier.

[0150] In certain embodiments, a pharmaceutical composition of the invention further comprises at least one additional pharmaceutically active agent other than a compound of the invention.

[0151] Pharmaceutical compositions of the invention can be prepared by combining one or more compounds of the invention with a pharmaceutically acceptable carrier and, optionally, one or more additional pharmaceutically active agents.

[0152] In certain embodiments, the pharmaceutical composition further comprises a saccharide.

[0153] In certain embodiments, the pharmaceutical composition further comprises a polyhydroxy acid.

[0154] In certain embodiments, the pharmaceutical composition further comprises a sugar alcohol.

[0155] As stated above, an effective amount refers to any amount that is sufficient to achieve a desired biological effect. Combined with the teachings provided herein, by choosing among the various active compounds and weighing factors such as potency, relative bioavailability, patient body weight, severity of adverse side-effects and mode of administration, an effective prophylactic or therapeutic treatment regimen can be planned which does not cause substantial unwanted toxicity and yet is effective to treat the particular subject. The effective amount for any particular application can vary depending on such factors as the disease or condition being treated, the particular compound of the invention being administered, the size of the subject, or the severity of the disease or condition. One of ordinary skill in the art can empirically determine the effective amount of a particular compound of the invention and/or other therapeutic agent without necessitating undue experimentation. A maximum dose may be used, that is, the highest safe dose according to some medical judgment. Multiple doses per day may be contemplated to achieve appropriate systemic levels of compounds. Appropriate systemic levels can be determined by, for example, measurement of the patient's peak or sustained plasma level of the drug. Dose and dosage are used interchangeably herein.

[0156] In certain embodiments, intravenous administration of a compound may typically be from about 300 mg/kg/day to about 1000 mg/kg/day. In one embodiment, intravenous administration of a compound may typically be from about 400 mg/kg/day to about 600 mg/kg/day. In one embodiment, intravenous administration of a compound may typically be from about 450 mg/kg/day to about 500 mg/kg/day.

[0157] Dosage may be adjusted appropriately to achieve desired drug levels, local or systemic, depending upon the mode of administration. For example, it is expected that intravenous administration would be from one order to several orders of magnitude lower dose per day. In the event that the response in a subject is insufficient at such doses, even higher doses (or effective higher doses by a different, more localized delivery route) may be employed to the extent that patient tolerance permits. Multiple doses per day are contemplated to achieve appropriate systemic levels of the compound.

[0158] For any compound described herein the therapeutically effective amount can be initially determined from animal models. A therapeutically effective dose can also be determined from human data for compounds which have been tested in humans and for compounds which are known to exhibit similar pharmacological activities, such as other related active agents. Higher doses may be required for parenteral administration. The applied dose can be adjusted based on the relative bioavailability and potency of the administered compound. Adjusting the dose to achieve maximal efficacy based on the methods described above and other methods as are well-known in the art is well within the capabilities of the ordinarily skilled artisan.

[0159] The formulations of the invention can be administered in pharmaceutically acceptable solutions, which may routinely contain pharmaceutically acceptable concentrations of salt, buffering agents, preservatives, compatible carriers, adjuvants, and optionally other therapeutic ingredients.

[0160] For use in therapy, an effective amount of the compound can be administered to a subject by any mode that delivers the compound to the desired surface. Administering a pharmaceutical composition may be accomplished by any means known to the skilled artisan. Routes of administration include but are not limited to intravenous, intramuscular, intraperitoneal, intravesical (urinary bladder), oral, subcutaneous, direct injection (for example, into a tumor or abscess), mucosal (e.g., topical to eye), inhalation, and topical.

[0161] For intravenous and other parenteral routes of administration, a compound of the invention can be formulated as a lyophilized preparation, as a lyophilized preparation of liposome-intercalated or -encapsulated active compound, as a lipid complex in aqueous suspension, or as a salt complex. Lyophilized formulations are generally reconstituted in suitable aqueous solution, e.g., in sterile water or saline, shortly prior to administration.

[0162] It will be understood by one of ordinary skill in the relevant arts that other suitable modifications and adaptations to the compositions and methods described herein are readily apparent from the description of the invention contained herein in view of information known to the ordinarily skilled artisan, and may be made without departing from the scope of the invention or any embodiment thereof. Having now described the present invention in detail, the same will be more clearly understood by reference to the following examples, which are included herewith for purposes of illustration only and are not intended to be limiting of the invention.

EXAMPLES

[0163] The invention is further described in the following examples, which do not limit the scope of the invention described in the claims.

[0164] Abbreviations used in the instant specification, particularly the schemes and examples, are as follows in Table A:

TABLE-US-00001 TABLE A Abbreviation Name ACN or MeCN acetonitrile AcOH glacial acetic acid aq aqueous conc. concentrated Cu(OAc).sub.2 copper(II) acetate DCM dichloromethane DIEA diisopropyl-ethyl amine DHP 3,4-Dihydropyran DMA dimethylaniline DMAP 4-dimethylaminopyridine DMF N,N-dimethylformamide DMSO dimethylsulfoxide Dppf or DPPF 1,1-Bis(diphenylphosphino)ferrocene EtOAc or EA ethyl acetate EtOH ethanol GCMS gas chromatography-mass spectrometry h or hr(s) hour or hours HPLC high performance liquid chromatography MeOH methanol MHz megahertz min minute or minutes LCMS Liquid chromatography mass spectrometry NBS N-Bromosuccinimide NIS N-iodosuccinimide NMR nuclear magnetic resonance Pd(PPh.sub.3).sub.4 tetrakis(triphenylphosphine)palladium Pd(OAc).sub.2 palladium (II) acetate PE petrolum ether PPh.sub.3 triphenylphosphine Pd(dppf)Cl.sub.2 1,1-bis(diphenylphosphino)ferrocene- palladium(II)dichloride FCC Flash column chromatography rt or RT room temperature TBS tert-Butyldimethylsilyl TEA or Et.sub.3N triethylamine TFA trifluoroacetic acid THF tetrahydrofuran TLC thin layer chromatography PTSA p-Toluenesulfonic acid

Example 1. Preparation of Compounds

[0165] ##STR00014##

[0166] Reagents and Conditions: (a) SOCl.sub.2, MeOH, 70 C.; (b) Boc.sub.2O, NaHCO.sub.3, dioxane, water, rt; (c) K.sub.2CO.sub.3, DMF, rt; (d) LiOH.Math.H.sub.2O, THF, water, rt; (e) HCl, EA, rt;

Preparation of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(3-hydroxyphenyl)propanoate (I-1.1)

[0167] Step A. To a mixture of 3-hydroxy-L-phenylalanine (5 g, 27.6 mmol) in MeOH (100 mL) at 0 C. was added SOCl.sub.2 (6.57 g, 55 mmol, 2 equiv) dropwise. The reaction mixture was heated at 70 C. for 4 h. The mixture was cooled to room temperature and concentrated under vacuum to give crude methyl (2S)-2-amino-3-(3-hydroxyphenyl)propanoate as a gum, which was used directly in the next step.

[0168] Step B. To a mixture of methyl (2S)-2-amino-3-(3-hydroxyphenyl)propanoate (1.37 g, 7.02 mmol, 1 equiv) and NaHCO.sub.3 (1.47 g, 17.5 mmol, 2.5 equiv) in dioxane (15 mL)/water (15 mL) was added di-tert-butyl dicarbonate (1.84 g, 8.4 mmol, 1.2 equiv) at room temperature. The reaction mixture was stirred overnight at room temperature. The resulting mixture was extracted with EtOAc (350 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated. The residue was purified by silica gel column chromatography, eluted with PE/EA (1:2) to afford methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(3-hydroxyphenyl)propanoate as a colorless liquid (2.02 g, 99%). LCMS (ESI): mass calcd. for C.sub.15H.sub.21NO.sub.5, 295.1; m/z found, 196.3 [M+HBoc].sup.+.

Preparation of (S)-3-(3-(boronomethoxy)phenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid (I-1.2)

[0169] Step C. To a mixture of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(3-hydroxyphenyl) propanoate (1 g, 3.39 mmol, 1 equiv) and K.sub.2CO.sub.3 (0.94 g, 6.772 mmol, 2 equiv) in DMF (10 mL) was added 2-(bromomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.96 g, 6.77 mmol, 2 equiv) dropwise. The reaction mixture was stirred at room temperature overnight, and then diluted with brine (40 mL). The mixture was extracted with EtOAc (520 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated. The crude product (1.8 g) was used in the next step directly without further purification. Note: 2-(iodomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane was also employed and provided an improved yield.

[0170] Step D. To a stirred solution of methyl (25)-2-[(tert-butoxycarbonyl)amino]-3-{3-[(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)methoxy]phenyl}propanoate (0.9 g, 2.07 mmol, 1 equiv) in THF (10 mL)/ water (5 mL) was added LiOH.Math.H.sub.2O (0.25 g, 6.01 mmol, 3 equiv) at room temperature. The reaction mixture was stirred at room temperature overnight. The resulting mixture was extracted with EtOAc (310 mL). The afforded aqueous layer was acidified to pH 5 with 1N HCl (aq.), the resulting mixture was extracted again with EtOAc (520 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated. This resulted in (S)-3-(3-(boronomethoxy)phenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid as a yellow oil (0.65 g, crude). LCMS (ESI): mass calcd. for C.sub.15H.sub.22BN07, 339.1; m/z found, 240.0 [M+HBoc].sup.+.

Preparation of (S)-2-amino-3-(3-(boronomethoxy)phenyl)propanoic acid; trifluoroacetic acid salt (I-1)

[0171] To a stirred solution of (S)-3-(3-(boronomethoxy)phenyl)-2-((tert-butoxycarbonyl)amino) propanoic acid (0.65 g, crude) in EtOAc (6 mL) was added 4M HCl in EtOAc (3 mL) at room temperature. The resulting mixture was stirred at room temperature overnight. Te resulting mixture was concentrated under reduced pressure. The crude product was purified by Prep-HPLC [with the following conditions (Column: Sunfire prep C18 column, 30*150 mm, 5 m; Mobile Phase A: Water (0.05% TFA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 4% B to 11% B in 7 min, 11% B; Wave Length: 254/220 nm; RT1 (min): 4.95)] to afford (S)-2-amino-3-(3-(boronomethoxy)phenyl)propanoic acid; trifluoroacetic acid as a white solid (70 mg, 9.90%). LCMS (ESI): mass calcd. for C.sub.10H.sub.14BNO.sub.5, 239.1; m/z found, 240.1 [M+H].sup.+. .sup.1H NMR (400 MHz, Deuterium Oxide) 7.23 (t, J =8.1 Hz, 1H), 6.91-6.83 (m, 1H), 6.83-6.73 (m, 2H), 4.17-4.00 (m, 1H), 3.70 (s, 2H), 3.19 (dd, J =14.5, 5.4 Hz, 1H), 3.04 (dd, J =14.5, 7.9 Hz, 1H).

##STR00015##

[0172] Reagents and Conditions: (a) (Trimethylsilyl)diazomethane (2M in hexanes), MeOH, toluene, 0 C.; (b) K.sub.2CO.sub.3, DMF, rt; (c) HCl (4M in EtOAc), rt; (d) LiOH, THF, H.sub.2O, rt.

Preparation of methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(2-fluoro-4-hydroxyphenyl) propanoate (I-2.1)

[0173] To a stirred mixture of (2S)-2-[(tert-butoxycarbonyl)amino]-3-(2-fluoro-4-hydroxyphenyl)propanoic acid (600 mg, 2.005 mmol, 1 equiv) in toluene (20 mL)/MeOH (5 mL) was added (trimethylsilyl)diazomethane (2M in hexanes) (2.406 mmol, 1.20 mL, 1.2 equiv) at 0 C. under nitrogen atmosphere. The resulting mixture was stirred under nitrogen atmosphere at room temperature for 0.5 h. The reaction was monitored by LCMS. The reaction was quenched by the addition of AcOH (0.5 mL) at room temperature, and then concentrated under reduced pressure. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% NH.sub.3.Math.H.sub.2O), 55% to 65% gradient in 10 min; detector, UV 254 nm]. This resulted in methyl (2S)-2-Rtert-butoxycarbonyl)aminol-3-(2-fluoro-4-hydroxyphenyl)propanoate as a white solid (540 mg, 85.97%). LCMS (ESI): mass calcd. for C.sub.15H.sub.20FNO.sub.5, 313.13; m/z found, 312.0 [MH].sup..

Preparation of methyl (S)-((4-(2-((tert-butoxycarbonyl)amino)-3-methoxy-3-oxopropyl)-3-fluorophenoxy)methyl)boronic acid (I-2.2)

[0174] To a stirred mixture of methyl (2S)-2-Rtert-butoxycarbonyl)aminol-3-(2-fluoro-4-hydroxyphenyl)propanoate (540 mg, 1.723 mmol, 1 equiv) and K.sub.2CO.sub.3 (1190.95 mg, 8.615 mmol, 5 equiv) in DMF (10 mL) at room temperature was added 2-(iodomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2308.56 mg, 8.615 mmol, 5 equiv). The resulting mixture was stirred under nitrogen atmosphere at room temperature overnight. The reaction was monitored by LCMS. The resulting mixture was diluted with water, and extracted with EtOAc (340 mL). The combined organic layers were washed with brine (280 mL), and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% NH.sub.3.Math.H.sub.2O), 50% to 60% gradient in 10 min; detector, UV 254 nm]. This resulted in (S)-((4-(2-((tert-butoxycarbonyl)amino)-3-methoxy-3-oxopropyl)-3-fluorophenoxy)methyl) boronic acid as a white solid (590 mg, 92.18%). LCMS (ESI): mass calcd. for C.sub.16H.sub.23BFNO.sub.7, 371.16; m/z found, 272.1 [M+HBoc].sup.+.

Preparation of 4-[(2S)-2-amino-3-methoxy-3-oxopropyl]-3-fluorophenoxymethylboronic acid (I-2.3)

[0175] To a stirred mixture of methyl (S)-((4-(2-((tert-butoxycarbonyl)arnino)-3-methoxy-3-oxopropyl)-3-fluorophenoxy)rnethyl)boronic acid (100 mg, 0.221 mmol, 1 equiv) in EtOAc (2.0 mL) under nitrogen atmosphere at room temperature was added 4 M HCl in EtOAc (2.0 mL) dropwise. The resulting mixture was stirred at room temperature for 5 h, and then concentrated under vacuum. The residue was diluted with water, and basified to pH 8 with saturated NaHCO.sub.3 (aq.). The resulting mixture was extracted with EtOAc (230 mL). The combined organic layers were washed with brine (250 mL), and dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% NH.sub.3.Math.H.sub.2O), 20% to 30% gradient in 8 min; detector, UV 254 nm]. This resulted in 4-[(2S)-2-amino-3-methoxy-3-oxopropyl]-3-fluorophenoxymethylboronic acid as a white solid (340 mg, 96.38%). LCMS (ESI): mass calcd. for C.sub.11H.sub.15BFNO.sub.5, 271.1; m/z found, 272.2 [M+H].sup.+.

Preparation of (S)-2-amino-3-(4-(boronomethoxy)-2-fluorophenyl)propanoic acid; trifluoroacetic acid (I-2)

[0176] To a stirred mixture of 4-[(2S)-2-amino-3-methoxy-3-oxopropyl]-3-fluorophenoxymethylboronic acid (340 mg, 1.25 mmol, 1 equiv) in THF (3 mL)/H.sub.2O (1 mL) was added LiOH.Math.H.sub.2O (157.9 mg, 3.76 mmol, 3.00 equiv) at room temperature. The resulting mixture was stirred for 5 h at room temperature under nitrogen atmosphere. The reaction mixture was acidified to pH 6 with 1N HCl (aq.), and then concentrated under reduced pressure. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm]. The product was further purified by Prep-HPLC [with the following conditions (Column: XBridge Shield RP18 OBD Column, 30*150 mm, 5 m; Mobile Phase A: water (0.05% TFA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 4% B to 10% B in 7 min, 10% B; Wave Length: 254/220 nm; RT1(min): 5.92)]. This resulted in (S)-2-amino-3-(4-(boronomethoxy)-2-fluorophenyl)propanoic acid; trifluoroacetic acid (110 mg, 22.50%) as a white solid. LCMS (ESI): mass calcd. for C.sub.10H.sub.13BFNO.sub.5, 257.1; m/z found, 257.9 [M+H].sup.+, .sup.1H NMR (400 MHz, Deuterium Oxide) 7.17 (t, J=8.6 Hz, 1H), 6.80-6.72 (m, 2H), 4.06 (dd, J=7.5, 5.5 Hz, 1H), 3.75 (s, 2H), 3.24 (dd, J=14.8, 5.6 Hz, 1H), 3.06 (dd, J=14.8, 7.6 Hz, 1H).

##STR00016##

[0177] Reagents and Conditions: (a) 1,2-dibromoethane, K.sub.2CO.sub.3, 18-crown-6, 80 C.; (b) CuCl, Xantphos, B.sub.2Pin.sub.2, .sup.tBuOK, DMF, 50 C.; (c) LiOH.Math.H.sub.2O, MeOH, H.sub.2O, rt; (d) HCl, dioxane, rt; (e) NaIO.sub.4, HCl, THF, H.sub.2O, rt.

Preparation of methyl (S)-3-(4-(2-brornoethoxy)phenyl)-2-((tert-butoxycarbonyl)amino) propanoate (I-3.1)

[0178] To a stirred solution of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(4-hydroxyphenyl) propanoate (5 g, 16.930 mmol, 1 equiv) in dibromoethane (20 mL) at room temperature were added K.sub.2CO.sub.3 (16.00 g, 115.801 mmol, 6.84 equiv), 18-crown-6 (0.45 g, 1.693 mmol, 0.1 equiv). The resulting mixture was stirred under nitrogen atmosphere at 80 C. for 24 h. The mixture was allowed to cool down to room temperature and diluted with ethyl acetate (150 mL). Then the mixture was washed with water and brine, dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (5:1) to afford methyl (S)-3-(4-(2-bromoethoxy)phenyl)-2-((tert-butoxycarbonyl)amino)propanoate as a white solid (5.6 g, 82.22%). LCMS (ESI): mass calcd. for C.sub.17H.sub.24BrNO.sub.2401.1; m/z found, 302.0 [M-Boc+H].sup.+.

Preparation of methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ethoxy)phenyl)propanoate (I-3.2)

[0179] To a stirred solution of methyl (S)-3-[4-(2-bromoethoxy)phenyl]-2-[(tert-butoxycarbonyl) amino]propanoate (5.6 g, 13.921 mmol, 1 equiv) and bis(pinacolato)diboron (10.61 g, 41.762 mmol, 3 equiv) in DMF (110 mL) at room temperature were added CuCl (413.44 mg, 4.177 mmol, 0.3 equiv), Xantphos (2.41 g, 4.177 mmol, 0.3 equiv) and Potassium tert-butoxide (1 M in THF) (16.6 mL, 1.2 equiv). The resulting mixture was purged with N2 for 1 min, and then stirred under nitrogen atmosphere at 50 C. for 3 h. The mixture was diluted with EA (80 mL) and washed with water (40 mL) and brine (40 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under vacuum and the residue was purified by silica gel column chromatography, eluted with PE/EA (4:1) to afford methyl (S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[2-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-yl)ethoxy]phenyl} propanoate (2.8 g, 44.76%) as a colorless oil. LCMS (ESI): mass calcd. for C.sub.23H.sub.36BNO.sub.7, 449.3; m/z found, 350.2 [MBoc+H].sup.+.

Preparation of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ethoxy)phenyl)propanoic acid (I-3.3)

[0180] To a stirred solution of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ethoxy]phenyl}propanoate (1.08 g, 2.403 mmol, 1 equiv) in H.sub.2O (20 mL) and MeOH (40 mL) at room temperature was added LiOH.Math.H.sub.2O (302.6 mg, 7.21 mmol, 3 equiv) in portions. The resulting mixture was stirred at room temperature for 3 h. Then the mixture was acidified to pH 4 with 1N HCl (aq.) and extracted with EA (330 mL), dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduced pressure. The residue was used in the next step directly without further purification. LCMS (ESI): mass calcd. for C.sub.22H.sub.34BNO.sub.7, 435.2; m/z found, 336.2 [MBoc+H].sup.+.

Preparation of (S)-2-amino-3-(4-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ethoxy)phenyl) propanoic acid (I-3.4)

[0181] To a stirred solution of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ethoxy]phenyl}propanoic acid (920 mg, 2.11 mmol, 1 equiv) in dioxane (9 mL) was added HCl (gas) solution (9 mL, 4 M in dioxane) dropwise. The resulting mixture was stirred at room temperature for 1 h, and then concentrated under reduced pressure. The residue was used in the next step directly without further purification. LCMS (ESI): mass calcd. for C.sub.17H.sub.26BNO.sub.5, 335.2; m/z found, 336.2 [M+H].sup.+.

Preparation of (2S)-2-amino-3-{4[2-(dihydroxyboranyl)ethoxy]phenyl}propanoic acid (I-3)

[0182] To a stirred solution of (2S)-2-amino-3-{4-[2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ethoxy]phenyl}propanoic acid (800 mg, 2.39 mmol, 1 equiv) in THF (20.00 mL) and H.sub.2O (5.00 mL) at room temperature was added sodium periodate (1.53 g, 7.161 mmol, 3 equiv) in portions. The resulting mixture was stirred at room temperature for 5 min, andthen 2N HCl (aq.) (1.00 mL, 2.005 mmol, 0.84 equiv) was added.. The reaction mixture was stirred at room temperature for additional 2 h. After that, the mixture was filtered, and the filter cake was washed with water (25 mL). The filtrate was concentrated under reduced pressure. The residue was purified by HP-Flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 5% to 30% gradient in 20 min; detector, UV 220 nm]. This resulted in (2S)-2-amino-3-{4-[2-(dihydroxyboranyl)ethoxy]phenyl}propanoic acid as a white solid (150 mg, 24.12%). LCMS (ESI): mass calcd. for C.sub.11H.sub.16BNO.sub.5, 253.1; m/z found, 254.1 [M+H].sup.+; .sup.1H NMR (400 MHz, Deuterium Oxide) 7.17 (d, J=8.2 Hz, 2H), 6.92 (d, J=8.2 Hz, 2H), 4.13 (t, J=7.5 Hz, 2H), 3.87 (dd, J=7.6, 5.3 Hz, 1H), 3.14 (dd, J=14.6, 5.2 Hz, 1H), 2.99 (dd, J=14.7, 7.8 Hz, 1H), 1.26 (t, J=7.6 Hz, 2H).

##STR00017##

[0183] Reagents and Conditions: (a) K.sub.2CO.sub.3, DMF, rt; (b) HCl (4M in EtOAc), EtOAc, rt.

Preparation of tert-butyl (2S)-2-[(tert-butoxycarbonyl)amino]3-{4-[(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)methoxy]phenyl}propanoate (I-4.1)

[0184] To a stirred mixture of tert-butyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(4-hydroxyphenyl) propanoate (1 g, 2.964 mmol, 1 equiv) and K.sub.2CO.sub.3 (0.82 g, 5.928 mmol, 2 equiv) in DMF (10 mL) at room temperature was added 2-(bromomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.31 g, 5.928 mmol, 2 equiv). The resulting mixture was stirred at room temperature overnight. The resulting mixture was diluted with water, and then extracted with EtOAc (2200 mL). The combined organic layers were washed with brine (3300 mL), dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduced pressure to afford tert-butyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)methoxy] phenyl}propanoate as a light yellow oil (1.34 g, crude). LCMS (ESI): mass calcd. for C.sub.25H.sub.40BNO.sub.7, 477.4; m/z found, 378 [M+HBoc].sup.+.

Preparation of 2-amino-3-{4-[(dihydroxyboranyl)methoxy]phenyl}propanoic acid (I-4)

[0185] To a stirred mixture of tert-butyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)methoxy]phenyl}propanoate (670 mg, 1.40 mmol, 1 equiv) in EtOAc (5 mL) at room temperature was added HC.sub.1(g) solution (10 mL, 4M in EtOAc) dropwise. The resulting mixture was stirred at room temperature overnight. The precipitated solids were collected by filtration and washed with ethyl ether (35 mL). The crude product (358 mg) was purified by Prep-HPLC with the following conditions (Column: Atlantis Prep T3 OBD Column, 19*150mm, 5 m; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 25 mL/min; Gradient: 2% B to 6% B in 6 min, 6% B; Wave Length: 254/220 nm; RT1 (min): 4.57) to afford 2-amino-3-{4-[(dihydroxyboranyl)methoxy]phenyl}propanoic acid as a white solid (49.1 mg, 14.61%).

[0186] LCMS (ESI): mass calcd. for C.sub.10H.sub.14BNO.sub.5, 239.0; m/z found, 240.0 [M+H].sup.+. .sup.1H NMR (400 MHz, Deuterium Oxide) 7.17 (d, J=8.7 Hz, 2H), 6.93 (d, J=8.6 Hz, 2H), 3.87 (dd, J=7.8, 5.1 Hz, 1H), 3.75 (s, 2H), 3.15 (dd, J=14.7, 5.2 Hz, 1H), 2.99 (dd, J=14.7, 7.8 Hz, 1H).

##STR00018##

[0187] Reagents and Conditions: (a) Pd(OAc).sub.2, S-phos, K.sub.3PO.sub.4, dioxane, H.sub.2O, 90 C.; (b) EtOH, EDCI, DMAP, DCM, rt; (c) BH3-THF, H.sub.2O, rt; (d) LiOH, MeOH, H.sub.2O, rt; (e) TFA, DCM, rt.

Preparation of (S)-2-((tert-butoxycarbonyl)amino)-3-(3-vinylphenyl)propanoic acid (I-5.1)

[0188] To a stirred mixture of (2S)-3-(3-bromophenyl)-2-[(tert-butoxycarbonyl)amino]propanoic acid (2 g, 5.81 mmol, 1 equiv) and 2-ethenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.34 g, 8.72 mmol, 1.5 equiv) in dioxane (50 mL) and H.sub.2O (5 mL) at room temperature was added K.sub.3PO.sub.4 (4.93 g, 23.2 mmol, 4 equiv), S-phos (0.48 g, 1.162 mmol, 0.2 equiv) and Pd(OAc).sub.2 (0.13 g, 0.581 mmol, 0.1 equiv). The reaction mixture was purged with N.sub.2 for 1 min, and then stirred under nitrogen atmosphere at 90 C. for 8 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3:1) to afford (2S)-2-[(tert-butoxycarbonyl)amino]-3-(3-ethenylphenyl)propanoic acid (2.1 g, crude) as a brown oil. LCMS (ESI): mass calcd. for C.sub.16H.sub.21NO.sub.4, 291.1; m/z found, 192 [MBoc+H].sup.+.

Preparation of ethyl (S)-2-((tert-butoxycarbonyl)amino)-3-(3-vinylphenyl)propanoate (I-5.2)

[0189] To a stirred mixture of (2S)-2-[(tert-butoxycarbonyl)amino]-3-(3-ethenylphenyl)propanoic acid (2.2 g, 7.551 mmol, 1 equiv), EtOH (3.48 g, 75.510 mmol, 10 equiv) and DMAP (0.09 g, 0.755 mmol, 0.1 equiv) in DCM (60 mL) was at room temperature added EDCI (1.59 g, 8.306 mmol, 1.1 equiv). The reaction mixture was stirred at 25 C. for 8 h, and then concentrated under reduced pressure. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 80% gradient in 10 min; detector, UV 254/220 nm] to afford ethyl (2S)-2-[(tert-butoxycarbonyl) amino]-3-(3-ethenylphenyl)propanoate as a light yellow solid (1.5 g, 62.19%). LCMS (ESI): mass calcd. for C.sub.18H.sub.25NO.sub.4, 319.2; m/z found, 220.1 [MBoc+H].sup.+.

Preparation of (S)-(3-(2-((tert-butoxycarbonyl)amino)-3-ethoxy-3-oxopropyl)phenethyl) boronic acid (I-5.3)

[0190] To a stirred mixture of ethyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(3-ethenylphenyl) propanoate (800 mg, 2.505 mmol, 1 equiv) in THF (10 mL) at 0 C. was added borane solution (1 M in THF, 5 mL, 5.01 mmol, 2 equiv) dropwise. The reaction mixture was stirred at 25 C. for 2 h, and then quenched by the addition of water at 0 C. The mixture was stirred for additional 2 h at 25 C., and then concentrated under reduced pressure. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254/220 nm] to afford 2-{3-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-ethoxy-3-oxopropyl]phenyl} ethylboronic acid as a light yellow oil (600 mg, 65.59%). LCMS (ESI): mass calcd. for C.sub.18H.sub.28BNO.sub.6, 365.2; m/z found, 266.2 [MBoc+H].sup.+.

Preparation of (S)-3-(3-(2-boronoethyl)phenyl)-2-((tert-butoxycarbonyl)amino)Propanoic Acid (I-5.4)

[0191] To a stirred mixture of 2-{3-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-ethoxy-3-oxopropyl] phenyl}ethylboronic acid (600 mg, 1.64 mmol, 1 equiv) in MeOH (6 mL) and H.sub.2O (6 mL) was added LiOH.Math.H.sub.2O (344.70 mg, 8.215 mmol, 5 equiv). The mixture was stirred at 25 C. for 2 h, and then acidified to pH 4 with 2N HCl (aq.). The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254/220 nm] to afford (S)-3-(3-(2-boronoethyl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid as an off-white solid (580 mg, crude). LCMS (ESI): mass calcd. for C.sub.16H.sub.24BNO.sub.6, 337.2; m/z found, 238.1 [MBoc+H].sup.+.

Preparation of (S)-2-amino-3-(3-(2-boronoethyl)phenyl)propanoic acid (I-5)

[0192] To a stirred mixture of (S)-3-(3-(2-boronoethyl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid (480 mg, 1.42 mmol, 1 equiv) in DCM (10 mL) at room temperature was added TFA (1 mL) dropwise. The mixture was stirred at 25 C. for 2 h, and then concentrated under reduced pressure. The residue was purified by Prep-HPLC [with the following conditions (Column: XBridge Prep Phenyl OBD Column, 19*150 mm, 5 m; Mobile Phase A: Water (0.1% FA), Mobile Phase B: MeOH-HPLC; Flow rate: 25 mL/min; Gradient: 9% B to 10% B in 6 min, 10% B; Wave Length: 220 nm; RT1(min): 4.11)] to afford (2S)-2-amino-3-{3-[2-(dihydroxyboranyl)ethyl]phenyl}propanoic acid as an off-white solid (83.3 mg, 24.19%). LCMS (ESI): mass calcd. for C.sub.11H.sub.16BNO.sub.4, 237.1; m/z found, 238.0 [M+H].sup.+; .sup.1H NMR (400 MHz, Methanol-d.sub.4) 7.24 (t, J=7.5 Hz, 1H), 7.20-7.03 (m, 3H), 4.00-3.84 (m, 1H), 3.31-3.21 (m, 1H), 3.01 (dd, J=14.5, 8.3 Hz, 1H), 2.67 (t, J=8.1 Hz, 2H), 1.11 (t, J=8.2 Hz, 2H).

##STR00019## ##STR00020##

[0193] Reagents and Conditions: (a) 1,2-dibromoethane, K.sub.2CO.sub.3, 18-crown-6, 80 C.; (b) CuCl, Xantphos, B.sub.2Pin.sub.2, .sup.tBuOK, DMF, 50 C.; (c) LiOH.Math.H.sub.2O, THF, H.sub.2O, rt; (d) NaIO4, HCl, THF, H.sub.2O, rt; (e) HCl, EA, rt.

Preparation of Methyl (2S)-3-(4-(2-brornoethoxy)-2-fluorophenyl)-2-((tert-butoxycarbonyl) amino)propanoate (I-6.1)

[0194] To a stirred solution of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(2-fluoro-4-hydroxyphenyl)propanoate (I-2.1, 1.03 g, 3.28 mmol, 1 equiv) in 1,2-dibromoethane (27.2 mL) at room temperature were added K.sub.2CO.sub.3 (3.12 g, 22.62 mmol, 6.9 equiv) and 18-crown-6 (86.6 mg, 0.327 mmol, 0.1 equiv). The resulting mixture was stirred under nitrogen atmosphere at 80 C. for 24 h . The reaction was allowed to cool down to room temperature and diluted with ethyl acetate (80 mL). Then the mixture was washed with brine (350 mL), dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (4:1) to afford methyl (2S)-3-[4-(2-bromoethoxy)-2-fluorophenyl]-2-[(tert-butoxycarbonyl)amino]propanoate as a light yellow oil (1.17 g, 85.20%). LCMS (ESI): mass calcd. for C.sub.17H.sub.23BrNO.sub.5, 419.1; m/z found, 320 [MBoc+H].sup.+.

Preparation of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{2-fluoro-4-[2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ethoxy]phenyl}propanoate (I-6.2)

[0195] A mixture of CuCl (82.9 mg, 0.837 mmol, 0.3 equiv) and Xantphos (484.62 mg, 0.837 mmol, 0.3 equiv) in DMF (22 mL) was stirred under nitrogen atmosphere at room temperature for 0.5 h. Next, methyl (2S)-3-[4-(2-bromoethoxy)-2-fluorophenyl]-2-[(tert-butoxycarbonyl)amino] propanoate (1.17 g, 2.79 mmol, 1 equiv), bis(pinacolato)diboron (2.13 g, 8.372 mmol, 3 equiv) and potassium tert-butoxide (1 M in THF) (3.3 mL, 3.3 mmol, 1.2 equiv) were added. The resulting mixture was purged with N.sub.2 for 1 min, and then stirred under nitrogen atmosphere at 50 C. 3 h. The reaction was quenched with addition of water (80 mL) and the mixture was extracted with EA (360 mL). The combined organic layers were washed with brine (100 mL) and dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under vacuum and the residue was purified by silica gel column chromatography, eluted with PE/EA (4:1) to afford methyl (2S)-2-[tert-butoxycarbonyl)amino]-3-{2-fluoro-4-[2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ethoxy]phenyl}propanoate as a light yellow oil (587 mg, 44.96%). LCMS (ESI): mass calcd. for C.sub.23H.sub.35BFNO.sub.7, 467.3; m/z found, 368.2 [MBoc+H].sup.+.

Preparation of (S)-2-((tert-butoxycarbonyl)amino)-3-(2-fluoro-4-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ethoxy)phenyl)propanoic acid (I-6.3)

[0196] To a stirred solution of methyl (2S)-2-[tert-butoxycarbonyl)amino]-3-{2-fluoro-4-[2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ethoxy]phenyl}propanoate (587 mg, 1.25 mmol, 1 equiv) in H.sub.2O (15 mL) and THF (30 mL) at room temperature was added LiOH.Math.H.sub.2O (105.3 mg, 2.51 mmol, 2.00 equiv) in portions. The resulting mixture was stirred at room temperature for 3 h. The mixture was acidified to pH 4 with 1M HCl (aq.) and extracted with EA (320 mL). The combined organic layers were dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was used in the next step directly without further purification. LCMS (ESI): mass calcd. for C.sub.22H.sub.33BFNO.sub.7, 453.2; m/z found, 354 [MBoc+H].sup.+.

Preparation of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[2-(dihydroxyboranyl)ethoxy]-2-fluorophenyl}propanoic Acid (I-6.4)

[0197] To a stirred solution of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{2-fluoro-4-[2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ethoxy]phenyl}propanoic acid (560 mg, 1.235 mmol, 1 equiv) in THF (9.0 mL) and H.sub.2O (2.3 mL) at room temperature was added sodium periodate (792.46 mg, 3.705 mmol, 3.0 equiv) in portions. The resulting mixture was stirred at room temperature for 5 min. Then 2 N HCl (aq.) (0.49 mL, 0.988 mmol, 0.8 equiv,) was added to the above solution. The reaction was stirred at room temperature for additional 2 h. The mixture was filtered, the filter cake was washed with water (25 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 10% to 80% gradient in 15 min; detector, UV 220 nm]. This resulted in (2S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[2-(dihydroxyboranyl)ethoxy]-2-fluorophenyl}propanoic acid as a light yellow solid (280 mg, 61.08%). LCMS (ESI): mass calcd. for C.sub.16H.sub.23BFNO.sub.7, 371.2; m/z found, 272.1 [MBoc+H].sup.+.

Preparation of (2S)-2-amino-3-{4-[2-(dihydroxyboranyl)ethoxy]-2-fluorophenyl}propanoic acid (I-6)

[0198] To a stirred solution of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[2-(dihydroxyboranyl)ethoxy]-2-fluorophenyl}propanoic acid (280 mg, 0.755 mmol, 1 equiv) in EA (5 mL) at room temperature was added hydrogen chloride (5 mL, 20.000 mmol, 4 M in EA) dropwise. The resulting mixture was stirred at room temperature for 2 h. Then the mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC [with the following conditions (Column: YMC-Actus Triart C18, 30*150 mm, 5 m; Mobile Phase A: Water (0.05% TFA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 10% B to 26% B in 7 min, 26% B; Wave Length: 254/220 nm; RT1(min): 5.23)] to afford (2S)-2-amino-3-{4-[2-(dihydroxyboranyl)ethoxy]-2-fluorophenyl} propanoic acid as a white solid (65 mg, 31.76%). LCMS (ESI): mass calcd. for C.sub.11H.sub.15BFNO.sub.5, 271.1; m/z found, 272.1 [M+H].sup.+; .sup.1H NMR (400 MHz, Deuterium Oxide) 7.13 (t, J=8.6 Hz, 1H), 6.75-6.64 (m, 2H), 4.08 (t, J=7.6 Hz, 2H), 3.86 (dd, J=7.5, 5.6 Hz, 1H), 3.18 (dd, J=14.8, 5.4 Hz, 1H), 2.97 (dd, J=14.8, 7.8 Hz, 1H), 1.24 (t, J=7.5 Hz, 2H).

##STR00021##

[0199] Reagents and Conditions: (a) Pd(OAc).sub.2, SPhos, K.sub.3PO.sub.4, dioxane/H.sub.2O, 95 C.; (b) EtOH, EDCI, DMAP, DCM, rt; (c) BH.sub.3-THF, THF, rt; (d) LiOH, H.sub.2O, EtOH, rt; (e) TFA, DCM, rt.

Preparation of (2S)-2-[(tert-butoxycarbonyl)amino]-3-(4-ethenylphenyl)propanoic acid (I-7.1)

[0200] To a stirred mixture of (2S)-3-(4-bromophenyl)-2-[(tert-butoxycarbonyl)amino]propanoic acid (5 g, 14.526 mmol, 1 equiv) and 2-ethenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (3.36 g, 21.789 mmol, 1.5 equiv) in 1,4-dioxane (100 mL) and H.sub.2O (10 mL) at room temperature were added K3PO4 (12.33 g, 58.104 mmol, 4 equiv), SPhos (1192.71 mg, 2.905 mmol, 0.2 equiv) and Pd(OAc)2 (0.33 g, 1.453 mmol, 0.1 equiv). The resulting mixture was purged with N.sub.2 for 1 min, and then stirred under nitrogen atmosphere at 95 C. overnight. The resulting mixture was diluted with water (50 mL), and extracted with EtOAc (3100 mL). The resulted aqueous layer was acidified to pH 5 with 2N HCl (aq.), and extracted again with EtOAc (3100 mL). The combined organic layers were washed with brine (2100 mL), and dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% TFA), 10% to 80% gradient in 15 min; detector, UV 254/220 nm]. This resulted in (2S)-2-[(tert-butoxycarbonyl)amino]-3-(4-ethenylphenyl)propanoic acid as a brown oil (5.12 g, crude). LCMS (ESI): mass calcd. for C.sub.16H.sub.21NO.sub.4, 291.1; m/z found, 290.0 [MH].sup..

Preparation of ethyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(4-ethenylphenyl)propanoate (I-7.2)

[0201] To a stirred mixture of (2S)-2-[(tert-butoxycarbonyl)amino]-3-(4-ethenylphenyl)propanoic acid (5.12 g, 17.574 mmol, 1 equiv) and DMAP (0.21 g, 1.757 mmol, 0.1 equiv) in EtOH (50 mL) and DCM (100 mL) at room temperature were added EDCI (3.71 g, 19.331 mmol, 1.1 equiv) in portions. The resulting mixture was stirred at room temperature for 8h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 30% to 100% gradient in 15 min; detector, UV 254/220 nm]. This resulted in ethyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(4-ethenylphenyl)propanoate as a brown oil (2.86 g, 50.95%). LCMS (ESI): mass calcd. for C.sub.18H.sub.25NO.sub.4, 319.1; m/z found, 220.0 [MBoc+H].sup.+.

Preparation of 2-{4-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-ethoxy-3-oxopropyl]phenyl} ethylboronic acid (I-7.3)

[0202] To a stirred mixture of ethyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(4-ethenylphenyl)propanoate (2.86 g, 8.95 mmol, 1 equiv) in THF (40 mL) under nitrogen atmosphere at 0 C. was added BH.sub.3-THF (17.91 mL, 17.908 mmol, 2 equiv, 1M in THF) dropwise . The resulting mixture was stirred at room temperature for 2h, and then quenched by the addition of water at 0 C. The resulting mixture was stirred at room temperature for additional 2h, and then concentrated under reduced pressure. The resulting mixture was extracted with CH.sub.2Cl.sub.2 (360 mL). The combined organic layers were washed with brine (2100 mL), and dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduced pressure. This resulted in 2-{4-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-ethoxy-3-oxopropyl]phenyl}ethylboronic acid as a light brown oil (3.37 g, crude). LCMS (ESI): mass calcd. for C.sub.18H.sub.28BNO.sub.6, 365.2; m/z found, 266.1 [MBoc+H].sup.+.

Preparation of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[2-(dihydroxyboranyl)ethyl]phenyl}propanoic acid (I-7.4)

[0203] To a stirred mixture of 2-{4-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-ethoxy-3-oxopropyl]phenyl}ethylboronic acid (3.85 g, 10.541 mmol, 1 equiv) in EtOH (60 mL) and H.sub.2O (6 mL) at room temperature was added LiOH.Math.H.sub.2O (2.21 g, 52.705 mmol, 5 equiv) in portions. The resulting mixture was stirred at room temperature for 2 h, and then diluted with water (100 mL). The resulting mixture was extracted with EtOAc (350 mL). The resulted aqueous layer was acidified to pH 4-5 with 2 N HCl (aq.). The resulting mixture was extracted with EtOAc (3100 mL). The combined organic layers were washed with brine (2100 mL), and dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduced pressure. This resulted in (2S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[2-(dihydroxyboranyl)ethyl]phenyl} propanoic acid as a light brown oil (3.37 g, 94.81%). LCMS (ESI): mass calcd. for C.sub.16H.sub.24BNO.sub.6, 337.1; m/z found, 238.1 [MBoc+H].sup.+.

[0204] Preparation of (2S)-2-amino-3-{4-[2-(dihydroxyboranyl)ethyl]phenyl}propanoic acid (I-7) To a stirred mixture of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[2-(dihydroxyboranyl) ethyl]phenyl}propanoic acid (900 mg, 2.669 mmol, 1 equiv) in DCM (18 mL) at room temperature was added TFA (4.00 mL) dropwise. The resulting mixture was stirred at room temperature for 2 h, and then concentrated under reduced pressure. The residue was purified by Prep-HPLC [with the following conditions (Column: XBridge Shield RP18 OBD Column, 30*150 mm, 5 m; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: to 6% B in 6 min, 6% B; Wave Length: 254/220 nm; RT1(min): 4.77)] to afford (2S)-2-amino-3-{4-[2-(dihydroxyboranyl)ethyl]phenyl}propanoic acid as a white solid (62.5 mg, 9.88%). LCMS (ESI): mass calcd. for C.sub.11H.sub.16BNO.sub.4, 237.1; m/z found, 237.9 [M+H]+; .sup.1H NMR (400 MHz, Methanol-d.sub.4) 7.26-7.14 (m, 4H), 3.75 (dd, J=8.8, 4.2 Hz, 1H), 3.30-3.22 (m, 1H), 2.96 (dd, J=14.6, 8.9 Hz, 1H), 2.65 (t, J=8.1 Hz, 2H), 1.09 (t, J=8.1 Hz, 2H).

##STR00022##

[0205] Reagents and Conditions: (a) SOCl.sub.2, EtOH, 60 C.; (b) Boc.sub.2O, Na.sub.2CO.sub.3, EtOH, rt; (c) Pd(OAc).sub.2, S-phos, K.sub.3PO.sub.4, dioxane, H.sub.2O, 90 C.; (d) BH.sub.3-THF, H.sub.2O, rt, then chiral resolution; (e) LiOH, MeOH, H.sub.2O, rt; (f) LiOH, MeOH, H.sub.2O, rt; (g) TFA, DCM, rt; (h) TFA, DCM, rt.

Preparation of ethyl 3-(4-bromo-2-fluorophenyl)-2-((tert-butoxycarbonyl)amino)propanoate (I-8.1)

[0206] Step A. To a stirred mixture of 2-amino-3-(4-bromo-2-fluorophenyl)propanoic acid (2 g, 7.631 mmol, 1 equiv) in ethyl alcohol (30 mL) was added thionyl chloride (4.54 g, 38.155 mmol, 5 equiv) dropwise at room temperature. The mixture was stirred at 60 C. for 8 h, and then concentrated under reduced pressure. The residue was basified to pH 8 with saturated Na.sub.2CO.sub.3 (aq.), and then extracted with DCM (3100 mL). The combined organic layers were washed with brine (2100 mL), dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduced pressure to afford ethyl 2-amino-3-(4-bromo-2-fluorophenyl)propanoate as a light yellow solid (2.5 g, crude), which was used directly in the next step.

[0207] Step B. To a stirred mixture of ethyl 2-amino-3-(4-bromo-2-fluorophenyl)propanoate (2.1 g, 7.24 mmol, 1 equiv) and Na.sub.2CO.sub.3 (3.07 g, 28.95 mmol, 4 equiv) in EtOH (50 mL) at room temperature was added di-tert-butyl dicarbonate (3.16 g, 14.48 mmol, 2 equiv). The reaction mixture was stirred at 25 C. for 8 h, and then concentrated under reduced pressure. The residue was diluted with water, and extracted with DCM (3100 mL). The combined organic layers were washed with brine (2200 mL), dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduced pressure to afford ethyl 3-(4-bromo-2-fluorophenyl)-2-[(tert-butoxycarbonyl) amino]propanoate as a light yellow solid (4.6 g, crude). LCMS (ESI): mass calcd. for C.sub.16H.sub.21BrFNO.sub.4, 389.1; m/z found, 290.0 [MBoc+H].sup.+.

[0208] Preparation of ethyl 2-((tert-butoxycarbonyl)amino)-3-(2-fluoro-4-vinylphenyl)propanoate (I-8.2)

[0209] To a stirred mixture of ethyl 3-(4-bromo-2-fluorophenyl)-2-[(tert-butoxycarbonyl) amino]propanoate (4.2 g, 10.762 mmol, 1 equiv) and 2-ethenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.49 g, 16.143 mmol, 1.5 equiv) in dioxane (80 mL) and H.sub.2O (8 mL) at room temperature was added K.sub.3PO.sub.4 (9.14 g, 43.048 mmol, 4 equiv), S-Phos (0.88 g, 2.152 mmol, 0.2 equiv) and Pd(OAc)2 (0.24 g, 1.076 mmol, 0.1 equiv). The reaction mixture was purged with N.sub.2 for 1 min, and then stirred under nitrogen atmosphere at 90 C. for 8 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (5:1) to afford ethyl 2-[(tert-butoxycarbonyl)amino]-3-(4-ethenyl-2-fluorophenyl)propanoate as a light yellow solid (2.1 g, 57.83%). LCMS (ESI): mass calcd. for C.sub.18H.sub.24FNO.sub.4, 337.2; m/z found, 238.2 [MBoc+H].sup.+.

Preparation of 2-{4-[(25)-2-[(tert-butoxycarbonyl)amino]-3-ethoxy-3-oxopropyl]-3-fluorophenyl}ethylboronic acid (1-8.3)and 2-{4-[(2R)-2-[(tert-butoxycarbonyl) amino]-3-ethoxy-3-oxopropyl]-3-fluorophenyl}ethylboronic acid (I-8.3)

[0210] To a stirred mixture of ethyl 2-[(tert-butoxycarbonyl)amino]-3-(4-ethenyl-2-fluorophenyl) propanoate (2 g, 5.928 mmol, 1 equiv) in THF (40 mL) at 0 C. was added borane solution (1M in THF, 5.9 mL, 11.86 mmol, 2 equiv) dropwise. The mixture was stirred at 25 C. for 2 h, and then quenched by the addition of water at 0 C. The mixture was stirred at 25 C. for additional 2 h, and then concentrated under reduced pressure. The residue was purified by Prep-HPLC [with the following conditions (Column: Sunfire prep C18 column, 30*150 mm, 5)..tm; Mobile Phase A: Water (0.05% TFA), Mobile Phase B: MeOHPreparative; Flow rate: 60 mL/min; Gradient: 55% B to 70% B in 11 min, 70% B; Wave Length: 254/220 nm; RT1(min): 9.48)] to afford 2-(4-{2-[(tert-butoxycarbonyl)amino]-3-ethoxy-3-oxopropyl}-3-fluorophenyl)ethyl-boronic acid (560 mg, 24.65%) as a colorless oil.

[0211] Then, the product (560 mg) was separated by Prep-SFC with the following conditions (Column: CHIRALCEL AY-H, 2*25 cm, 5 m; Mobile Phase A: CO.sub.2, Mobile Phase B: EtOH-HPLC; Flow rate: 40 mL/min; Gradient: isocratic 10% B; Column Temperature ( C.): 35; Back Pressure(bar): 100; Wave Length: 220 nm; RT1 (min): 2.87; RT2 (min): 3.77; Sample Solvent: MeOHPreparative; Injection Volume: 0.2 mL) to afford 2-14-[(2S*)-2-[(tert-butoxycarbonyl)amino]-3-ethoxy-3-oxopropyl]-3- fluorophenyljethylboronic acid (I-8.3, 132 mg, 23.57%, ee>99%, 1.sup.st isomer on HPLC) as an off-white solid and 2-14-[(2R*)-2-[(tert-butoxycarbonyl)amino]-3-ethoxy-3-oxopropyl]- 3-fluorophenyljethylboronic acid (I-8.3,135 mg, 24.11%, ee>97%, 2.sup.nd isomer on HPLC) as an off-white solid. LCMS (ESI): mass calcd. for C.sub.18H.sub.27BFNO.sub.6, 383.2; m/z found, 284.0 [MBoc+H].sup.+.

Preparation of (S*)-3-(4-(2-boronoethyl)-2-fluorophenyl)-2-((tert-butoxycarbonyl)amino) propanoic acid (I-8.4)

[0212] To a stirred mixture of 2-{4-[(2S*)-2-[(tert-butoxycarbonyl)amino]-3-ethoxy-3-oxopropyl]-3-fluorophenyl}ethylboronic acid (130 mg, 0.339 mmol, 1 equiv) in MeOH (2 mL) and H.sub.2O (2 mL) at room temperature was added LiOH.Math.H.sub.2O (71.12 mg, 1.695 mmol, 5 equiv). The mixture was stirred at 25 C. for 2 h, and then acidified to pH 4 with 2N HCl (aq.). The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, ACN in Water (0.1% FA), 10% to 60% gradient in 10 min; detector, UV 254/220 nm] to afford (2S *)-2-[(tert-butoxycarbonyl)amino]-3-{4-[2-(dihydroxyboranyl)ethyl]-2-fluorophenyl}propanoic acid (100 mg, crude) as an off-white solid. LCMS (ESI): mass calcd. for C.sub.16H.sub.23BFNO.sub.6, 355.2; m/z found, 255.95 [MBoc+H].sup.+.

[0213] Preparation of (R*)-3-(4-(2-boronoethyl)-2-fluorophenyl)-2-((tert-butoxycarbonyl)amino) propanoic acid (I-8.4)

[0214] To a stirred mixture of 2-{4-[(2R*)-2-[(tert-butoxycarbonyl)amino]-3-ethoxy-3-oxopropyl]-3-fluorophenyl}ethylboronic acid (130 mg, 0.339 mmol, 1 equiv) in MeOH (2 mL) and H.sub.2O (2 mL) at room temperature was added LiOH.Math.H.sub.2O (71.12 mg, 1.695 mmol, 5 equiv). The mixture was stirred at 25 C. for 2 h, and then acidified to pH 4 with 2N HCl (aq.). The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 10% to 60% gradient in 10 min; detector, UV 254/220 nm] to afford (2R*)-2-[(tert-butoxycarbonyl)amino]-3-{4-[2-(dihydroxyboranyl)ethyl]-2-fluorophenyl}propanoic acid as an off-white solid(100 mg, crude). LCMS (ESI): mass calcd. for C.sub.16H.sub.23BFNO.sub.6, 355.2; m/z found, 256.0 [MBoc+H].sup.+.

Preparation of (S*)-2-amino-3-(4-(2-boronoethyl)-2-fluorophenyl)propanoic acid (I-8)

[0215] To a stirred mixture of (2S*)-2-[(tert-butoxycarbonyl)amino]-3-{4-[2-(dihydroxyboranyl)ethyl]-2-fluorophenyl}propanoic acid (100 mg, 0.282 mmol, 1 equiv) in DCM (3 mL) was added TFA(0.5 mL) dropwise at room temperature. The mixture was stirred at 25 C. for 2 h, and then concentrated under reduced pressure. The residue was purified by Prep-HPLC [with the following conditions (Column: XBridge Shield RP18 OBD Column, 19*150 mm, 5 m; Mobile Phase A: Water (0.1% FA), Mobile Phase B: MeOH-HPLC; Flow rate: 25 mL/min; Gradient: 5% B to 17% B in 6 min, 17% B; Wave Length: 220 nm; RT1(min): 4.72)] to afford (2S*)-2-amino-3-{4-[2-(dihydroxyboranyl)ethyl]-2-fluorophenyl} propanoic acid; trifluoroacetic acid (56.9 mg, 54.76%) as an off-white solid. LCMS (ESI): mass calcd. for C.sub.11H.sub.15BFNO.sub.4, 255.2; m/z found, 255.9 [M+H].sup.+; .sup.1H NMR (400 MHz, Methanol-d.sub.4) 7.22 (t, J=7.8 Hz, 1H), 7.07-6.87 (m, 2H), 4.07-3.87 (m, 1H), 3.41-3.32 (m, 1H), 3.05 (dd, J=14.6, 7.9 Hz, 1H), 2.67 (t, J=8.1 Hz, 2H), 1.10 (t, J=8.2 Hz, 2H).

Preparation of (R*)-2-amino-3-(4-(2-boronoethyl)-2-fluorophenyl)propanoic acid (I-9)

[0216] To a stirred mixture of (2R*)-2-[(tert-butoxycarbonyl)amino]-3-{4-[2-(dihydroxyboranyl)ethyl]- 2-fluorophenyl}propanoic acid (110 mg, 0.310 mmol, 1 equiv) in DCM (3 mL) was added TFA (0.5 mL) dropwise at room temperature. The mixture was stirred at 25 C. for 2 h, and then concentrated under reduced pressure. The residue was purified by Prep-HPLC [with the following conditions (Column: XBridge Shield RP18 OBD Column, 19*150 mm, 5 m; Mobile Phase A: Water (0.1% FA), Mobile Phase B: MeOH-HPLC; Flow rate: 25 mL/min; Gradient: 5% B to 20% B in 7 min, 20% B; Wave Length: 220 nm; RT1 (min): 4.85)] to afford (2R*)-2-amino-3-{4-[2-(dihydroxyboranyl)ethyl]-2-fluorophenyl} propanoic acid; trifluoroacetic acid as an off-white solid (38.3 mg, 33.18%). LCMS (ESI): mass calcd. for C.sub.11H.sub.15BFNO.sub.4, 255.2; m/z found, 256.15 [M+H].sup.+; .sup.1H NMR (400 MHz, Methanol-d.sub.4) 7.23 (t, J=7.8 Hz, 1H), 7.06-6.87 (m, 2H), 3.91-3.71 (m, 1H), 3.41-3.32 (m, 1H), 2.99 (dd, J=14.5, 8.5 Hz, 1H), 2.67 (t, J=8.0 Hz, 2H), 1.10 (t, J=8.0 Hz, 2H).

##STR00023##

[0217] Reagents and Conditions: (a) SOCl.sub.2, MeOH, 50 C.; (b) Boc.sub.2O, TEA, DCM, THF; (c) K.sub.2CO.sub.3, DMF, 2-(iodomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane; (d) HCl in EA, EA; (e) LiOH H.sub.2O, THF, H.sub.2O.

Preparation of methyl (2S)-2-amino-3-(3-fluoro-4-hydroxyphenyl)propanoate (I-10.1)

[0218] To a stirred mixture of 3-fluorotyrosine (1 g, 5.021 mmol, 1 equiv) in methanol (20 mL) under nitrogen atmosphere at 0 C. was added thionyl chloride (3.58 g, 30.126 mmol, 6 equiv) dropwise. The resulting mixture was stirred under nitrogen atmosphere at 50 C. for 4 h, and then concentrated. The residue was basified to pH 8 with saturated NaHCO.sub.3 (aq.). The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% NH.sub.3.Math.H.sub.2O), 30% to 40% gradient in 8 min; detector, UV 254/220 nm]. This resulted in methyl (2S)-2-amino-3-(3-fluoro-4-hydroxyphenyl)propanoate as a white solid (920 mg, 85.95%). LCMS (ESI): mass calcd. for Cioth2FN03, 213.1; m/z found, 214.3 [M+H].sup.+.

Preparation of methyl (2S)-2-[(tert-butoxycarbonyl)Amino]-3-(3-fluoro-4-hydroxyphenyl) propanoate (I-10.2)

[0219] To a stirred mixture of methyl (2S)-2-amino-3-(3-fluoro-4-hydroxyphenyl)propanoate (860 mg, 4.034 mmol, 1 equiv) and TEA (612.26 mg, 6.051 mmol, 1.5 equiv) in DCM/THF (10 mL/10 mL) under nitrogen atmosphere at room temperature was added di-tert-butyl dicarbonate (1056.40 mg, 4.841 mmol, 1.2 equiv) . The resulting mixture was stirred under nitrogen atmosphere at room temperature overnight, and then concentrated under reduced pressure. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% NH.sub.3.Math.H.sub.2O), 30% to 50% gradient in 8 min; detector, UV 254/220 nm]. This resulted in methyl (2S)-2-Rtert-butoxycarbonyl)aminol-3-(3-fluoro-4-hydroxyphenyl)propanoate as a white solid (924 mg, 73.11%). LCMS (ESI): mass calcd. for C.sub.15H.sub.20FNO.sub.5, 313.1; m/z found, 314.2 [M+H].sup.+.

Preparation of 4-[(2S)-2-[(tert-butoxycarbonyl)Amino]-3-methoxy-3-oxopropyl]-2-fluorophenoxyrnethylboronic acid (I-10.3)

[0220] To a stirred mixture of methyl (2S)-2-Rtert-butoxycarbonyl)aminol-3-(3-fluoro-4-hydroxyphenyl)propanoate (924 mg, 2.949 mmol, 1 equiv) and K.sub.2CO.sub.3 (2.04 g, 14.745 mmol, 5 equiv) in DMF (18 mL) under nitrogen atmosphere at room temperature was added 2-(iodomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (3.95 g, 14.744 mmol, 5.00 equiv) . The resulting mixture was stirred under nitrogen atmosphere at room temperature overnight. The reaction mixture was purified directly by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 30% to 50% gradient in 8 min; detector, UV 254/220 nm]. This resulted in 4-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxy-3-oxopropyl]-2- fluorophenoxymethylboronic acid as a white solid (630 mg, 57.56%). LCMS (ESI): mass calcd. for C.sub.16H.sub.23FNO.sub.7, 371.2; m/z found, 272.1 [M+HBoc].sup.+.

Preparation of 4-[(2S)-2-amino-3-methoxy-3-oxopropyl]-2-fluorophenoxymethylboronic acid (I-10.4)

[0221] To a stirred mixture of 4-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxy-3-oxopropyl]-2-fluorophenoxymethylboronic acid (900 mg, 2.43 mmol, 1 equiv) in EtOAc (10 mL) under nitrogen atmosphere at room temperature was added hydrogen chloride (5 mL, 4 M in EtOAc) dropwise . The resulting mixture was stirred under nitrogen atmosphere at room temperature for 3 h, and then concentrated under reduced pressure. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 15% to 40% gradient in 7 min; detector, UV 254/220 nm]. This resulted in 4-[(2S)-2-amino-3-methoxy-3-oxopropyl]-2-fluorophenoxymethylboronic acid (550 mg, 83.68%) as a white solid. LCMS (ESI): mass calcd. for C.sub.11H.sub.15BFNO.sub.5, 271.1; m/z found, 272.3 [M+H].sup.+.

Preparation of (2S)-2-amino-3-{4-[(dihydroxyboranyl)methoxy]-3-fluorophenyl}propanoic acid (I-10)

[0222] To a stirred mixture of 4-[(2S)-2-amino-3-methoxy-3-oxopropyl]-2-fluorophenoxymethylboronic acid (550 mg, 2.029 mmol, 1 equiv) in THF/H.sub.2O (8 mL/8 mL) under nitrogen atmosphere at room temperature was added LiOH.Math.H.sub.2O (255.43 mg, 6.087 mmol, 3 equiv) . The resulting mixture was stirred under nitrogen atmosphere at room temperature for 4 h, and then acidified to pH 6 with 2N HCl (aq.). The resulting mixture was concentrated under vacuum. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 0% to 5% gradient in 8 min; detector, UV 254/220 nm]. The product was further purified by Prep-HPLC [with the following conditions (Column: Atlantis Prep T3 OBD Column, 19*150mm 5 m; Mobile Phase A: Water (0.1% TFA), Mobile Phase B: ACN; Flow rate: 25 mL/min; Gradient: 8% B to 22% B in 7 min, 22% B; Wave Length: 254/220 nm; RT1(min): 5.04)]. This resulted in (2S)-2-amino-3-{4-[(dihydroxyboranyl)methoxy]-3-fluorophenyl} propanoic acid; trifluoroacetic acid as a white solid (68 mg, 13.04%). LCMS (ESI): mass calcd. for C.sub.10H.sub.13BFNO.sub.5, 257.1; m/z found, 258.1 [M+H].sup.+. .sup.1H NMR (400 MHz, Deuterium Oxide) 7.12-6.96 (m, 3H), 3.95 (dd, J=7.7, 5.3 Hz, 1H), 3.85 (s, 2H), 3.17 (dd, J=14.7, 5.3 Hz, 1H), 3.03 (dd, J=14.7, 7.7 Hz, 1H).

##STR00024##

[0223] Reagents and Conditions: (a) SOCl.sub.2, MeOH, 50 C.; (b) Boc.sub.2O, DCM, TEA; (c) K.sub.2CO.sub.3, DMF, 2-(iodomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane; (d) HCl in EA, EA; (e) LiOH.Math.H.sub.2O, THF, H.sub.2O.

Preparation of methyl (2S)-2-amino-3-(4-fluoro-3-hydroxyphenyl)propanoate (I-11.1)

[0224] To a stirred mixture of (2S)-2-amino-3-(4-fluoro-3-hydroxyphenyl)propanoic acid hydrochloride (900 mg, 3.819 mmol, 1 equiv) in methanol (18 mL) under nitrogen atmosphere at 0 C. was added thionyl chloride (5 mL) . The resulting mixture was stirred under nitrogen atmosphere at 50 C. overnight, and then concentrated under vacuum. The residue was basified to pH 8 with saturated Na.sub.2CO.sub.3 (aq.). The mixture was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol/L NH4HCO3), 30% to 40% gradient in 10 min; detector, UV 254/220 nm]. This resulted in methyl (2S)-2-amino-3-(4-fluoro-3-hydroxyphenyl)propanoate as a white solid (680 mg, 83.50%). LCMS (ESI): mass calcd. for C.sub.10H.sub.12FNO.sub.3, 213.1; m/z found, 214.1 [M+H].sup.+.

Preparation of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(4-fluoro-3-hydroxyphenyl)propanoate (I-11.2)

[0225] To a stirred mixture of methyl (2S)-2-amino-3-(4-fluoro-3-hydroxyphenyl)propanoate (800 mg, 3.752 mmol, 1 equiv) and TEA (759.39 mg, 7.504 mmol, 2 equiv) in DCM (16 mL) under nitrogen atmosphere at room temperature was added di-tert-butyl dicarbonate (982.70 mg, 4.502 mmol, 1.2 equiv) . The resulting mixture was stirred under nitrogen atmosphere at room temperature overnight, and then concentrated under reduced pressure. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% NH.sub.3.Math.H.sub.2O), 50% to 60% gradient in 7 min; detector, UV 254/220 nm]. This resulted in methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(4-fluoro-3-hydroxyphenyl)propanoate as a white solid (297 mg, 25.26%). LCMS (ESI): mass calcd. for C.sub.15H.sub.20FN05, 313.1; m/z found, 312.2 [MH].sup..

Preparation of 5-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxy-3-oxopropyl]-2-fluoro-phenoxyrnethylboronic acid (I-11.3)

[0226] To a stirred mixture of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(4-fluoro-3-hydroxyphenyl)propanoate (488 mg, 1.557 mmol, 1 equiv) and K.sub.2CO.sub.3 (1.08 g, 7.785 mmol, 5 equiv) in DMF (9 mL) under nitrogen atmosphere at room temperature was added 2-(iodomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.09 g, 7.785 mmol, 5 equiv) . The resulting mixture was stirred under nitrogen atmosphere at room temperature overnight. The reaction mixture was purified directly by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 30% to 55% gradient in 8 min; detector, UV 254/220 nm]. This resulted in 5-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxy-3-oxopropyl]-2- fluorophenoxymethylboronic acid as a white solid (434 mg, 75.07%). LCMS (ESI): mass calcd. for C.sub.16H.sub.23BFNO.sub.7, 371.2; m/z found, 272.3 [M+HBoc].sup.+.

Preparation of 5-[(2S)-2-amino-3-methoxy-3-oxopropyl]-2-fluorophenoxymethylboronic acid (I-11.4)

[0227] To a stirred mixture of 5-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxy-3-oxopropyl]-2-fluorophenoxymethylboronic acid (290 mg, 0.781 mmol, 1 equiv) in EtOAc (3 mL) under nitrogen atmosphere at room temperature was added HCl (1 mL, 4 N in EtOAc) dropwise . The resulting mixture was stirred under nitrogen atmosphere at room temperature overnight. The resulting mixture was concentrated under vacuum. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 0% to 5% gradient in 10 min; detector, UV 254/220 nm]. This resulted in 5-[(2S)-2-amino-3-methoxy-3-oxopropyl]-2-fluorophenoxymethylboronic acid as a white solid (167 mg, 78.86%). LCMS (ESI): mass calcd. for C.sub.11H.sub.15BFNO.sub.3, 271.1; m/z found, 272.2 [M+H].sup.+.

Preparation of (2S)-2-amino-3-{3-[(dihydroxyboranyl)methoxy]-4-fluorophenyl}propanoic acid (I-11)

[0228] To a stirred mixture of 5-[(2S)-2-amino-3-methoxy-3-oxopropyl]-2-fluorophenoxymethylboronic acid (317 mg, 1.170 mmol, 1 equiv) in THF/H.sub.2O (6 mL/3 mL) under nitrogen atmosphere at room temperature was added LiOH.Math.H.sub.2O (147.22 mg, 3.510 mmol, 3 equiv). The resulting mixture was stirred under nitrogen atmosphere at room temperature for 3 h, and then acidified to pH 6 with 2N HCl (aq.). The resulting mixture was concentrated under vacuum. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 0% to 5% gradient in 8 min; detector, UV 254/220 nm. The product was further purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30*150 mm, 5 m; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 5% B to 20% B in 7 min, 20% B; Wave Length: 254/220 nm; RT1(min): 3.58)]. This resulted in (2S)-2-amino-3-{3-[(dihydroxyboranyl)methoxy]-4-fluorophenyl}propanoic acid as a white solid (60 mg, 19.04%). LCMS (ESI): mass calcd. for C.sub.10H.sub.13BFNO.sub.5, 257.1; m/z found, 258.2 [M+H].sup.+. .sup.1H NMR (400 MHz, Deuterium Oxide) 6 7.07 (dd, J=11.5, 8.3 Hz, 1H), 6.98 (d, J=8.3 Hz, 1H), 6.82-6.75 (m, 1H), 3.93-3.87 (m, 1H), 3.82 (s, 2H), 3.17 (dd, J=14.6, 5.2 Hz, 1H), 3.02 (dd, J=14.6, 7.9 Hz, 1H).

##STR00025##

[0229] Reagents and Conditions: (a) SOCl.sub.2, MeOH, 50 C.; (b) Boc.sub.2O, NaHCO.sub.3, dioxane, H.sub.2O; (c) K.sub.2CO.sub.3, DMF, 2-(iodomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane; (d) LiOH H.sub.2O, THF, H.sub.2O; (e) HCl in EA, EA.

Preparation of methyl (2S)-2-amino-3-(2-fluoro-3-hydroxyphenyl)propanoate (I-12.1)

[0230] To a stirred mixture of (2S)-2-amino-3-(2-fluoro-3-hydroxyphenyl)propanoic acid (800 mg, 4.016 mmol, 1 equiv) in methanol (16 mL) under nitrogen atmosphere at 0 C. was added thionyl chloride (8 mL) dropwise . The resulting mixture was stirred under nitrogen atmosphere at 50 C. for 4 h. The resulting mixture was concentrated. The residue was basified to pH 8 with saturated NaHCO.sub.3 (aq.). The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% NH.sub.3.Math.H.sub.2O), 10% to 20% gradient in 8 min; detector, UV 254/220 nm]. This resulted in methyl (2S)-2-amino-3-(2-fluoro-3-hydroxyphenyl)propanoate as a white solid (790 mg, 92.25%). LCMS (ESI): mass calcd. for C.sub.10H.sub.12FNO.sub.3, 213.1; m/z found, 214.3 [M+H].sup.+.

Preparation of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(2-fluoro-3-hydroxyphenyl) propanoate (I-12.2)

[0231] To a stirred mixture of methyl (2S)-2-amino-3-(2-fluoro-3-hydroxyphenyl)propanoate (2.72 g, 12.8 mmol, 1 equiv) and NaHCO.sub.3 (2.68 g, 31.9 mmol, 2.5 equiv) in dioxane/H.sub.2O (27 mL/27 mL) under nitrogen atmosphere at room temperature was added di-tert-butyl dicarbonate (4.18 g, 19.1 mmol, 1.5 equiv) in portions . The resulting mixture was stirred under nitrogen atmosphere at room temperature for 5 h. The resulting mixture was filtered, and the filtering pad was washed with MeOH (310 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% NH.sub.3.Math.H.sub.2O), 30% to 50% gradient in 8 min; detector, UV 254/220 nm]. This resulted in methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(2-fluoro-3-hydroxyphenyl)propanoate as a white solid (1.914 g, 47.88%). LCMS (ESI): mass calcd. for C.sub.15H.sub.20FN05, 313.1; m/z found, 312.2 [MH].sup..

Preparation of 3-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxy-3-oxopropyl]-2-fluorophenoxymethylboronic acid (1-12.3)

[0232] To a stirred mixture of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(2-fluoro-3-hydroxyphenyl)propanoate (286 mg, 0.913 mmol, 1 equiv) and K.sub.2CO.sub.3 (630.76 mg, 4.565 mmol, 5 equiv) in DMF (5 mL) under nitrogen atmosphere at room temperature was added 2-(iodomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.2 g, 4.565 mmol, 5 equiv) . The resulting mixture was stirred under nitrogen atmosphere at room temperature overnight. The reaction mixture was purified directly by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 30% to 45% gradient in 8 min; detector, UV 254/220 nm]. This resulted in 3-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxy-3-oxopropyl]-2- fluorophenoxymethylboronic acid as a white solid (210 mg, 61.98%). LCMS (ESI): mass calcd. for C.sub.16H.sub.23BFNO.sub.7, 371.2; m/z found, 272.1 [M+HBoc].sup.+.

Preparation of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{3-[(dihydroxyboranyl)methoxy]-2-fluorophenyl}propanoic acid (I-12.4)

[0233] To a stirred mixture of 3-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxy-3-oxopropyl]-2-fluorophenoxymethylboronic acid (302 mg, 0.814 mmol, 1 equiv) in THF/H.sub.2O (6 mL/3 mL) was under nitrogen atmosphere at room temperature added LiOH.Math.H.sub.2O (58.46 mg, 2.442 mmol, 3 equiv) . The resulting mixture was stirred under nitrogen atmosphere at room temperature for 3 h, and then acidified to pH 6 with 1N HCl (aq.). The resulting mixture was concentrated under vacuum. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 20% to 35% gradient in 8 min; detector, UV 254/220 nm]. This resulted in (2S)-2-[(tert-butoxycarbonyl)amino]-3-{3-[(dihydroxyboranyl)methoxy]-2-fluorophenyl}propanoic acid as a white solid (210 mg, 72.27%). LCMS (ESI): mass calcd. for C.sub.15H.sub.21BFNO.sub.7, 357.1; m/z found, 356.3 [MH].sup..

Preparation of (2S)-2-amino-3-{3-[(dihydroxyboranyl)methoxy]-2-fluorophenyl}propanoic acid; trifluoroacetic acid (I-12)

[0234] To a stirred mixture of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{3-[(dihydroxyboranyl)methoxy]-2-fluorophenyl}propanoic acid (210 mg, 0.588 mmol, 1 equiv) in EtOAc (2 mL) under nitrogen atmosphere at room temperature was added HCl (2 mL, 4 M in EtOAc) . The resulting mixture was stirred under nitrogen atmosphere at room temperature for 2 h, and then concentrated under vacuum. The residue was purified by Prep-HPLC [with the following conditions (Column: XBridge Prep Amide OBD Column, 19*150 mm, 5 m; Mobile Phase A: Water (0.05% TFA), Mobile Phase B: ACN; Flow rate: 25 mL/min; Gradient: 95% B to 83% B in 2 min, 83% B to 60% B in 10 min, 60% B; Wave Length: 220 nm; RT1(min): 7.79)]. This resulted in (2S)-2-amino-3-{3-[(dihydroxyboranyl)methoxy]-2-fluorophenyl}propanoic acid; trifluoroacetic acid as a white solid (55 mg, 36.39%). LCMS (ESI): mass calcd. for C.sub.10H.sub.13BFNO.sub.5, 257.1; m/z found, 258.1 [M+H].sup.+. .sup.1H NMR (400 MHz, Deuterium Oxide) 7.10-6.98 (m, 2H), 6.85-6.75 (m, 1H), 4.06 (dd, J=7.6, 5.8 Hz, 1H), 3.81 (s, 2H), 3.29 (dd, J=14.5, 5.7 Hz, 1H), 3.10 (dd, J=14.7, 7.7 Hz, 1H).

##STR00026##

[0235] Reagents and Conditions: (a) SOCl.sub.2, MeOH, 50 C.; (b) Boc.sub.2O, NaHCO.sub.3, dioxane, H.sub.2O, rt; (c) 2-(bromomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane, K.sub.2CO.sub.3, DMF, rt; (d) LiOH.Math.H.sub.2O, THF, H.sub.2O, rt; (e) HCl in EtOAc (4 M), EtOAc, rt.

Preparation of methyl (S)-2-amino-3-(3-fluoro-5-hydroxyphenyl)propanoate (1-13.1)

[0236] To a stirred mixture of (2S)-2-amino-3-(3-fluoro-5-hydroxyphenyl)propanoic acid (1 g, 5.02 mmol, 1 equiv) in MeOH (20 mL) under nitrogen atmosphere at 0 C. was added SOCl.sub.2 (10 mL) dropwise . The resulting mixture was stirred under nitrogen atmosphere at 50 C. overnight, and then concentrated under vacuum. The residue was basified to pH 8 with saturated NaHCO.sub.3 (aq.). The resulting mixture was extracted with EtOAc (350 mL). The combined organic layers were washed with brine (280 mL), dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was used in the next step directly without further purification. LCMS (ESI): mass calcd. for C.sub.10H.sub.15FNO.sub.3, 213.1; m/z found, 214.3 [M+H].sup.+.

Preparation of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(3-fluoro-5-hydroxyphenyl) propanoate (I-13.2)

[0237] To a stirred mixture of methyl (S)-2-amino-3-(3-fluoro-5-hydroxyphenyl)propanoate (1.5 g, 7.035 mmol, 1 equiv) and NaHCO.sub.3 (0.89 g, 10.553 mmol, 1.5 equiv) in dioxane/H.sub.2O (15 mL/15 mL) under nitrogen atmosphere at room temperature was added Boc.sub.2O (1.54 g, 7.035 mmol, 1 equiv) . The resulting mixture was stirred under nitrogen atmosphere at room temperature for 3 h., and then diluted with water. The resulting mixture was extracted with EtOAc (350 mL). The combined organic layers were washed with brine (280 mL), dried over anhydrous Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 50% to 80% gradient in 15 min; detector, UV 254/220 nm]. This resulted in methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(3-fluoro-5-hydroxyphenyl)propanoate as a white solid (750 mg, 34.02%). LCMS (ESI): mass calcd. for C.sub.15H.sub.20FNO.sub.5, 313.1; m/z found, 312.2 [MH].sup..

Preparation of 3-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxy-3-oxopropyl]-5-fluoro-phenoxymethylboronic acid (I-13.3)

[0238] To a stirred mixture of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-(3-fluoro-5-hydroxyphenyl)propanoate (630 mg, 2.01 mmol, 1 equiv) and K.sub.2CO.sub.3 (1.39 g, 10.1 mmol, 5 equiv) in DMF (8 mL) under nitrogen atmosphere at room temperature was added 2-(iodomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.69.g, 10.055 mmol, 5 equiv) . The resulting mixture was stirred under nitrogen atmosphere at room temperature overnight. The resulting mixture was filtered, the filter cake was washed with MeOH (35 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 55% to 60% gradient in 10 min; detector, UV 254/220 nm]. This resulted in 3-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxy-3-oxopropyl]-5-fluorophenoxymethylboronic acid as a white solid (520 mg, 69.68%). LCMS (ESI): mass calcd. for C.sub.16H.sub.23BFNO.sub.7, 371.2; m/z found, 272.1 [M+HBoc].sup.+.

Preparation of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{3-[(dihydroxyboranyl)methoxy]-5-fluorophenyl}propanoic acid(I-13.4)

[0239] To a stirred mixture of 3-[(2S)-2-[(tert-butoxycarbonyl)amino]-3-methoxy-3-oxopropyl]-5-fluorophenoxymethylboronic acid (888 mg, 2.392 mmol, 1 equiv) in THF/H.sub.2O (16 mL/8 mL) under nitrogen atmosphere at room temperature was added LiOH.Math.H.sub.2O (301.16 mg, 7.177 mmol, 3.00 equiv) . The resulting mixture was stirred under nitrogen atmosphere at room temperature for 3 h, and then neutralized to pH 7 with 2N HC.sub.1. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 35% to 45% gradient in 10 min; detector, UV 254/220 nm]. This resulted in (2S)-2-[(tert-butoxycarbonyl)amino]-3-{3-[(dihydroxyboranyl)methoxy]-5-fluorophenyl}propanoic acid as a white solid (630 mg, 73.73%). LCMS (ESI): mass calcd. for C.sub.15H.sub.21BFNO.sub.7, 357.1; m/z found, 356.1 [MH].sup..

Preparation of (2S)-2-amino-3-[3-[(dihydroxyboranyl)methoxy]-5-fluorophenyl]propanoic acid (I-13)

[0240] To a stirred mixture of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{3-[(dihydroxyboranyl)methoxy]-5-fluorophenyl}propanoic acid (210 mg, 0.588 mmol, 1 equiv) in EtOAc (2 mL) under nitrogen atmosphere at room temperature was added HCl (1 mL, 4 N in EtOAc) dropwise . The resulting mixture was stirred under nitrogen atmosphere at room temperature for 3 h, and then concentrated under vacuum. The residue was neutralized to pH 7 with NaOH (aq.). The mixture was purified by reversed-phase flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 3% to 5% gradient in 8 min; detector, UV 254/220 nm]. The product was further purified by Prep-HPLC [with the following conditions (Column: XBridge Shield RP18 OBD Column, 30*150 mm, 5 m; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 50 mL/min; Gradient: 2% B to 18% B in 8 min, 18% B; Wave Length: 254/220 nm; RT1(min): 7.28)]. This resulted in (25)-2-amino-3-{3-[(dihydroxyboranyl)methoxy]-5-fluorophenyl}propanoic acid as a white solid (105 mg, 69.48%). LCMS (ESI): mass calcd. for C.sub.10H.sub.13BFNO.sub.5, 257.1; m/z found, 258.2 [M+H].sup.+. .sup.1H NMR (400 MHz, Deuterium Oxide) 6.69-6.61 (m, 2H), 6.59 (d, J=9.2 Hz, 1H), 3.91 (dd, J=7.9, 5.2 Hz, 1H), 3.72 (s, 2H), 3.16 (dd, J=14.5, 5.3 Hz, 1H), 3.00 (dd, J=14.5, 8.0 Hz, 1H).

##STR00027##

[0241] .sup.aReagents and Conditions: (a) CataCXium A Pd G3, Cs.sub.2CO.sub.3, dioxane/H2O, 100 C.; (b) LiOH.Math.H.sub.2O, THF, water, rt; (c) HCl, dioxane, rt;

Preparation of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{3-[1-(4,4,5,5 -tetramethyl- 1,3,2-dioxaborolan-2-yl)cyclopropyl]phenyl}propanoate (I-14.1)

[0242] To a stirred mixture of methyl (2S)-3-(3-bromophenyl)-2-[(tert-butoxycarbonyl)amino]propanoate (731.06 mg, 2.041 mmol, 1.20 equiv) and 4,4,5,5-tetramethyl-2-[1-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclopropyl]-1,3,2-dioxaborolane (500 mg, 1.701 mmol, 1.00 equiv) in dioxane (16 mL)/H.sub.2O (1.6 mL) was added CataCXium A Pd G3 (123.85 mg, 0.170 mmol, 0.1 equiv) and Cs2CO3 (1662.29 mg, 5.103 mmol, 3 equiv). The resulting mixture was purged with nitrogen for 1 min, and then stirred under nitrogen atmosphere at 100 C. for 5 h. The resulting mixture was concentrated under vacuum. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 60% to 80% gradient in 15 min; detector, UV 220 nm]. This resulted in methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{3-[1-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclopropyl]phenyl}propanoate as a light yellow oil (240 mg, 31.69%). LCMS (ESI): mass calcd. for C.sub.24H.sub.36BNO.sub.6, 445.3; m/z found, 446.3 [M+H].sup.+.

Preparation of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{3-[1-(dihydroxyboranyl) cyclopropyl] phenyl}propanoic acid (1-14.2)

[0243] To a stirred mixture of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{3-[1-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclopropyl]phenyl}propanoate (240 mg, 0.539 mmol, 1 equiv) in H.sub.2O (2.5 mL)/THF (2.5 mL) was added LiOH.Math.H.sub.2O (67.84 mg, 1.617 mmol, 3 equiv). The resulting mixture was stirred at room temperature for 3 h. The mixture was acidified to pH 7 with 2 M HCl (aq.) and concentrated under vacuum. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 40% to 60% gradient in 10 min; detector, UV 220 nm]. This resulted in (2S)-2-[(tert-butoxycarbonyl)amino]-3-{3-[1-(dihydroxyboranyl)cyclopropyl]phenyl}propanoic acid as a colorless oil (120 mg, 63.77%). LCMS (ESI): mass calcd. for C.sub.17H.sub.24BNO.sub.6, 349.2; m/z found, 250.1 [MBoc+H].sup.+

Preparation of (2S)-2-amino-3-{3-[1-(dihydroxyboranyl)cyclopropyl] phenyl}propanoic acid (I-14)

[0244] To a stirred mixture of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{3-[1-(dihydroxyboranyl) cyclopropyl] phenyl}propanoic acid (150 mg, 0.430 mmol, 1 equiv) in dioxane (2.0 mL) was added a 1,4-dioxane solution (4 mL) of HCl (0.7 mL) in dropwise. The resulting mixture was stirred at room temperature for 3 h, and then concentrated under vacuum. The crude product was purified by Prep-HPLC [with the following conditions (Column: XBridge Prep Phenyl OBD Column, 19*150 mm, 5 m; Mobile Phase A: Water (0.1% FA), Mobile Phase B: MeOH-HPLC; Flow rate: 25 mL/min; Gradient: 17% B to 42% B in 7 min, 42% B; Wave Length: 220 nm; RT1(min): 5.11)] to afford (2S)-2-amino-3-{3-[1-(dihydroxyboranyl)cyclopropyl] phenyl}propanoic acid as a white solid (27 mg, 25.24%). LCMS (ESI): mass calcd. for C.sub.12H.sub.16BNO.sub.4, 249.1; m/z found, 250.2 [M+H].sup.+. .sup.1H NMR (400 MHz, Deuterium Oxide) 7.27-7.18 (m, 2H), 7.16 (s, 1H), 7.04 (d, J=7.0 Hz, 1H), 3.89 (dd, J=8.0, 5.2 Hz, 1H), 3.17 (dd, J=14.5, 5.2 Hz, 1H), 2.98 (dd, J=14.5, 8.3 Hz, 1H), 1.05-0.97 (m, 2H), 0.87-0.80 (m, 2H).

##STR00028##

[0245] Reagents and Conditions: (a) CataCXium A Pd G3, Cs.sub.2CO.sub.3, dioxane/H20, 100 C.; (b) LiOH.Math.H.sub.2O, THF, water, rt; (c) HCl, dioxane, rt.

Preparation of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{3-[1-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclopropyl]phenyl}propanoate (I-15.1)

[0246] To a stirred mixture mixture of methyl (2S)-3-(4-bromophenyl)-2-[(tert-butoxycarbonyl) amino]propanoate (292.42 mg, 0.816 mmol, 1.2 equiv) and 4,4,5,5-tetramethyl-2-[1-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclopropyl]-1,3,2-dioxaborolane (200 mg, 0.680 mmol, 1.00 equiv) in dioxane (6 mL)/H.sub.2O (0.6 mL) was added CataCXium A Pd G3 (74.31 mg, 0.102 mmol, 0.15 equiv) and Cs.sub.2CO.sub.3 (54.57 mg, 0.168 mmol, 3 equiv). The resulting mixture was purged with nitrogen for 1 min, and then stirred under nitrogen atmosphere at 80 Cfor 5 h. The resulting mixture was concentrated under vacuum. The residue was purified by reverse flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 60% to 80% gradient in 20 min; detector, UV 220 nm]. This resulted in methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[1-(4,4,5,5-tetramethyl-1,3 ,2-dioxaborolan-2-yl)cyclopropyl]phenyl }propanoate as a light yellow oil (110 mg, 36.31%). LCMS (ESI): mass calcd. for C.sub.24H.sub.36BNO.sub.6, 445.3; m/z found, 468.2 [M+Na].sup.+.

Preparation of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[1-(dihydroxyboranyl) cyclopropyl] phenyl}propanoic acid (I-15.2)

[0247] To a stirred mixture of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[1-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclopropyl]phenyl}propanoate (100 mg, 0.225 mmol, 1 equiv) in H.sub.2O (1.0 mL)/THF (1.0 mL) was added LiOH.Math.H.sub.2O (28.26 mg, 0.675 mmol, 3 equiv). The resulting mixture was stirred at room temperature for 3 h. The mixture was acidified to pH 7 with 2 M HCl (aq.) and then concentrated under vacuum. The residue was purified by reversed-phase flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 40% to 60% gradient in 15 min; detector, UV 220 nm]. This resulted in (2S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[1-(dihydroxyboranyl)cyclopropyl]phenyl}propanoic acid as a colorless oil (70 mg, 89.28%). LCMS (ESI): mass calcd. for C.sub.17H.sub.24BNO.sub.6, 349.2; m/z found, 250.1 [MBoc+H].sup.+.

Preparation of (2S)-2-amino-3-{4-[1-(dihydroxyboranyl)cyclopropyl]phenyl}propanoic acid (I-15)

[0248] To a stirred mixture of (2S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[1-(dihydroxyboranyl) cyclopropyl] phenyl}propanoic acid (70 mg, 0.200 mmol, 1 equiv) in dioxane (1.2 mL) was added a 1,4-dioxane solution (4M) of HCl (0.40 mL) dropwise. The resulting mixture was stirred at room temperature for 3 h, and then concentrated under vacuum. The crude product was purified by Prep-HPLC [with the following conditions (Column: Xselect CSH C18 OBD Column 30*150 mm 5 m; Mobile Phase A: Water (0.1%FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 7% B to 19% B in 7 min, 19% B; Wave Length: 220 nm; RT1 (min): 6.78)] to afford (2S)-2-amino-3-{4-[1-(dihydroxyboranyl)cyclopropyl]phenyl}propanoic acid as a white solid (31 mg, 62.09%). LCMS (ESI): mass calcd. for C.sub.12H.sub.16BNO.sub.4, 249.1; m/z found, 250.1 [M+H].sup.+. .sup.1H NMR (400 MHz, Deuterium Oxide) 7.24 (d, J=7.9 Hz, 2H), 7.12 (d, J=7.9 Hz, 2H), 3.91-3.81 (m, 1H), 3.14 (dd, J=14.5, 5.3 Hz, 1H), 2.99 (dd, J=14.5, 7.8 Hz, 1H), 1.05-0.92 (m, 2H), 0.88-0.74 (m, 2H).

##STR00029##

[0249] Reagents and Conditions: (a) Zn, 12, Pd2(dba)3, S-Phos, DMF, 45 C.; (b) K.sub.2CO.sub.3, DMF, rt; (c) LiOH. H.sub.2O, THF, H.sub.2O, rt; (d) HCl, dioxane, rt.

Preparation of methyl (2R)-2-[(tert-butoxycarbonyl)amino]-3-(3-hydroxyphenyl)propanoate (I-16-1)

[0250] A mixture of Zn (3.4 g, 52.02 mmol, 3 equiv) and I.sub.2 (0.22 g, 1.73 mmol, 0.1 equiv) in DMF (30 mL) was stirred under nitrogen atmosphere at room temperature for 5 min. To the above mixture was added a solution of methyl (2S)-2-[(tert-butoxycarbonyl)amino]-3-iodopropanoate (6.85 g, 20.81 mmol, 1.2 equiv) in DMF (30 mL) slowly, followed by the addition of I.sub.2 (0.22 g, 1.73 mmol, 0.1 equiv). The resulting mixture was stirred under nitrogen atmosphere at room temperature for 30 min. Then, to the mixture was added a suspension of 3-bromophenol (3 g, 17.34 mmol, 1 equiv), S-Phos (0.71 g, 1.73 mmol, 0.1 equiv) and Pd2(dba).sub.3 (0.79 g, 0.86 mmol, 0.05 equiv) in DMF (10 mL). The reaction was stirred under nitrogen atmosphere at 45 C. overnight. The reaction was quenched with water (20 mL), diluted with EA (80 mL), and filtered through Celite. The filtrate was washed with brine (320 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated. The residue was purified by reversed-phase flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in water (0.1% FA), 40% to 60% gradient in 20 min; detector, UV 254 nm] to provide the title product as a yellow oil (2.7g, 52.7%). LCMS (ESI): mass calcd. for C.sub.15H.sub.21NO.sub.5, 295.1; m/z found, 296.1 [M+H].sup.+.

Preparation of methyl (2R)-2-[(tert-butoxycarbonyl)amino]-3-{3-[(4,4,5,5-tetrarnethyl-1,3,2-dioxaborolan-2-yl)methoxy]phenyl}propanoate (I-16-2)

[0251] To a stirred mixture of methyl (2R)-2-[(tert-butoxycarbonyl)amino]-3-(3-hydroxyphenyl) propanoate (2.7 g, 9.14 mmol, 1 equiv) and K.sub.2CO.sub.3 (1.9 g, 13.71 mmol, 1.5 equiv) in DMF (20 mL) was added 2-(iodomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.9 g, 10.97 mmol, 1.2 equiv) slowly. The resulting mixture was stirred under nitrogen atmosphere at room temperature overnight. The reaction was quenched with water (10 mL), and extracted with EtOAc (330 mL). The combined organic layers were washed with brine (220 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in water (10 mmol/L NH4HCO3), 10% to 40% gradient in 60 min; detector, UV 254 nm] to afford the title product as a brown oil (675 mg, 17.0%). LCMS (ESI): mass calcd. for C.sub.24H.sub.35BFNO.sub.6, 435.2; m/z found, 434.2 [MH].sup..

Preparation of (2R)-2-[(tert-butoxycarbonyl)amino]-3-{3-[(dihydroxyboranyl)methoxy] phenyl}propanoic acid (I-16-3)

[0252] To a stirred mixture of methyl (2R)-2-[(tert-butoxycarbonyl)aminol-3-{3-[(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)methoxy]phenyl}propanoate (391 mg, 0.90 mmol, 1 equiv) in THF (8 mL)/H.sub.2O (4 mL) was added LiOH.Math.H.sub.2O (75.6 mg, 1.80 mmol, 2 equiv). The resulting mixture was stirred at room temperature for 3 h. Then the mixture was acidified to pH 5 with 2N HCl (aq.), and extracted with EA (320 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography [with the following conditions: column, C18 silica gel; mobile phase, MeCN in water (0.1% FA), 10% to 50% gradient in 30 min; detector, UV 220 nm] to afford the title product as a light yellow solid (215 mg, 70.6%). LCMS (ESI): mass calcd. for C.sub.15H.sub.22BNO.sub.7, 339.2; m/z found, 337.8 [MH].sup..

Preparation of (2R)-2-amino-3-{3-[(dihydroxyboranyl)methoxy]phenyl}propanoic acid (16)

[0253] To a stirred solution of (2R)-2-[(tert-butoxycarbonyl)amino]-3-{3-[(dihydroxyboranyl) methoxy]phenyl}propanoic acid (200 mg, 0.59 mmol, 1 equiv) in dioxane (2 mL) was added HC.sub.1 (2 mL, 4 mol/L in dioxane) slowly. The resulting mixture was stirred under nitrogen atmosphere at room temperature for 2 h. Then the mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC [with the following conditions: Column: XBridge Prep Amide OBD Column, 19*150 mm, 5 m; Mobile Phase A: Water (0.05%TFA), Mobile Phase B: ACN; Flow rate: 60 mL/min mL/min; Gradient: 95% B to 83% B in 10 min; Wave Length: 254nm/220nm nm; RT1(min): 8.32] to afford the title product as a white solid (57.3 mg, 40.5%). LCMS (ESI): mass calcd. for C.sub.10H.sub.14BNO.sub.5, 239.10; m/z found, 240.10 [M+H].sup.+; .sup.1H NMR (400 MHz, Deuterium Oxide) 7.33-7.21 (m, 1H), 6.93-6.86 (m, 1H), 6.86-6.79 (m, 2H), 4.29-4.16 (m, 1H), 3.80-3.64 (m, 2H), 3.32-3.16 (m, 1H), 3.16-3.01 (m, 1H).

Example 2. Cell Uptake Assays

Cell Culture

1.1 Thawing Method

[0254] 1.1.1 15 mL of cell culture medium was placed into a T225 flask. [0255] 1.1.2 The flask was placed in a humidified 37 C., 5% CO.sub.2 incubator for 15 minutes to allow medium to equilibrate to the proper pH and temperature. [0256] 1.1.3 The vial was removed from liquid nitrogen and thawed rapidly by placing at 37 C. in a water bath with gentle agitation for 1-2 minutes and then decontaminated by wiping with 70% ethanol before opening in a Class II biological safety cabinet. [0257] 1.1.4 The vial contents were transferred dropwise into 10 mL of cell culture medium in a sterile 15 mL conical tube. [0258] 1.1.5 The cells were centrifuged at 1,300 rpm for 5 minutes. [0259] 1.1.6 The supernatant was aspirated and re-suspended the cells and transferred the cells into T225 flask containing cell culture medium.

1.2 Propagation Method

[0260] 1.2.1 Culture medium were renewed every 2-3 days. [0261] 1.2.2 Keep the flask in a humidified 37 C., 5% CO.sub.2 incubator

Assay Procedures

2.1 Cell Seeding

[0262] Cells were harvested and diluted in culture medium to the designated concentration. Then the cells were cultured in T25 flasks, one T25 flask for one sample.
2.2 Formulation of test compounds [0263] 2.2.1 L-Boronophenylalanine (L-BPA, 4-boron-L-phenylalanine, Purity: 95%, Chemical formula: C.sub.9H12BNO4, MW: 209.01, CAS: 76410-58-7) and fructose were dissolved in PBS at a molar ratio of 1:5, and then a 1.27 M ratio of 1 N NaOH is added. The mixture was stirred until L-BPA is completely dissolved, and the pH value was titrated to 7.2-7.4 with 1 N HC.sub.1. [0264] 2.2.2 For test compounds other than BPA, stock solutions were prepared of each compound in DMSO or fructose formulation at a concentration of 20 mM (such that final % DMSO in the cellular assay medium=0.5%)
2.3 Compound treatment [0265] 2.3.1 The cells were seeded overnight (12-18 hours) to allow the cells to adhere well. [0266] 2.3.2 The stock solution of each compound was added to each T25 flask such that the final concentration for each compound is 100 uM (0.1 mM). [0267] 2.3.3 The flasks were placed in a 37 C., 5% CO.sub.2 incubator for 1 hour, 4 hours and 24 hours. [0268] 2.3.4 After treatment for 1 hour, 4 hour, and 24 hours, the cells were collected by using 0.25% Trypsin with 0.53 mM EDTA. [0269] 2.3.5 Centrifuged at 1,300 rpm for 5 min to collect cell pellets.

Results

[0270] The compounds disclosed herein were selectively taken up by the representative human cancer cell lines SAS (head & neck cancer), U87-MG (glioblastoma) and B6 (melanoma) relative to a representative normal human cell line (NIH-3T3). Compounds 1-3, 10-11, and 13-15, in addition to showing selective partitioning into cancer cells relative to normal cells, showed increased uptake in cancer cells relative to BPA.

TABLE-US-00002 TABLE 1 Cell uptake of compound 1 as compared to BPA 1 BPA Time Mean (ng/10.sup.6 cell) Time Mean (ng/10.sup.6 cell) (h) SAS B16F10 NIH-3T3 U87-MG (h) SAS B16F10 NIH-3T3 U87-MG 1.00 112 127 227 1.00 294 1 8 91.4 101 4.00 755 232 194 233 4.00 259 120 109 85.2 24.0 828 517 133 336 24.0 962 257 70.1 136 indicates data missing or illegible when filed

TABLE-US-00003 TABLE 2a Cell uptake of compound 2 as compared to BPA 2 BPA Time Mean (ng/10.sup.6 cell) Time Mean (ng/10.sup.6 cell) (h) SAS B16F10 NIH-3T3 U87-MG (h) SAS B16F10 NIH-3T3 U87-MG 1.00 1125 151 .5 123 1.00 294 108 91.4 101 4.00 1590 333 20 40 4.00 259 120 109 85.2 24.0 1595 881 205 626 24.0 962 2 7 70.1 136 indicates data missing or illegible when filed

TABLE-US-00004 TABLE 3 Cell uptake of compound 3 as compared to BPA 3 BPA Time Mean (ng/10.sup.6 cell) Time Mean (ng/10.sup.6 cell) (h) SAS B16F10 NIH-3T3 U87-MG (h) SAS B16F10 NIH-3T3 U87-MG 1.00 534 145 132 197 1.00 294 108 91.4 101 4.00 499 222 138 211 4.00 259 120 109 85.2 24.0 NA 117 7 183 24.0 962 257 70.1 136 indicates data missing or illegible when filed

TABLE-US-00005 TABLE 4 Cell uptake of compound 4 as compared to BPA 4 BPA Time Mean (ng/10.sup.6 cell) Time Mean (ng/10.sup.6 cell) (h) SAS B16F10 NIH-3T3 U87-MG (h) SAS B16F10 NIH-3T3 U87-MG 1.00 445 46.1 27.1 42.8 1.00 294 10 91.4 101 4.00 495 95.6 58.6 113 4.00 259 120 109 85.2 24.0 997 101 66.6 191 24.0 9 2 257 70.1 138 indicates data missing or illegible when filed

TABLE-US-00006 TABLE 5 Cell uptake of compound 5 as compared to BPA 5 BPA Time Mean (ng/10.sup.6 cell) Time Mean (ng/10.sup.6 cell) (h) SAS B16F10 NIH-3T3 U87-MG (h) SAS B16F10 NIH-3T3 U87-MG 1.00 1 3 12 171 1 1.00 2 7 126 105 156 4.00 225 157 77.2 1 4.00 185 1 9 2 1 7 24.0 214 262 0.3 12 24.0 1 3 332 9 156 indicates data missing or illegible when filed

TABLE-US-00007 TABLE 6 Cell uptake of compound 6 as compared to BPA 6 BPA Time Mean (ng/10.sup.6 cell) Time Mean (ng/10.sup.6 cell) (h) SAS B16F10 NIH-3T3 U87-MG (h) SAS B16F10 NIH-3T3 U87-MG 1.00 152 53.4 160 1.00 297 126 105 156 4.00 106 92.3 23.2 137 4.00 185 188 98.2 167 24.0 46.0 2 .0 NA 1 7 24.0 193 332 92 156 indicates data missing or illegible when filed

TABLE-US-00008 TABLE 7 Cell uptake of compound 8 as compared to BPA 8 BPA Time Mean (ng/10.sup.6 cell) Time Mean (ng/10.sup.6 cell) (h) SAS B16F10 NIH-3T3 U87-MG (h) SAS B16F10 NIH-3T3 U87-MG 1.00 44.6 26.9 14.7 16.7 1.00 163 102 69.7 9 .6 4.00 66.7 31.4 26.4 31.1 4.00 157 143 98.0 127 24.0 359 38. 3 .4 16.3 24.0 328 144 38.2 94.6 indicates data missing or illegible when filed

TABLE-US-00009 TABLE 8 Cell uptake of compound 9 as compared to BPA 9 BPA Time Mean (ng/10.sup.6 cell) Time Mean (ng/10.sup.6 cell) (h) SAS B16F10 NIH-3T3 U87-MG (h) SAS B16F10 NIH-3T3 U87-MG 1.00 129 67 34.4 90.1 1.00 163 102 69.7 9 4.00 103 66.7 43.0 1 7 4.00 157 143 9 127 24.0 211 119 37.2 74.6 24.0 328 144 38.2 94.6 indicates data missing or illegible when filed

TABLE-US-00010 TABLE 9 Cell uptake of compound 10 as compared to BPA 10 BPA Time Mean (ng/10.sup.6 cell) Time Mean (ng/10.sup.6 cell) (h) SAS B16F10 NIH-3T3 U87-MG (h) SAS B16F10 NIH-3T3 U87-MG 1.00 1285 23 81.9 295 1.00 782 252 97. 197 4.00 1850 442 204 673 4.00 11 234 10 317 24.0 14 0 814 12 681 24.0 1345 267 75.9 172 indicates data missing or illegible when filed

TABLE-US-00011 TABLE 10 Cell uptake of compound 11 as compared to BPA 11 BPA Time Mean (ng/10.sup.6 cell) Time Mean (ng/10.sup.6 cell) (h) SAS B16F10 NIH-3T3 U87-MG (h) SAS B16F10 NIH-3T3 U87-MG 1.00 529 50.6 32.4 113 1.00 1 8 148 110 410 4.00 454 121 92.8 322 4.00 3 0 227 129 345 24.0 999 202 122 309 24.0 49 315 133 178 indicates data missing or illegible when filed

TABLE-US-00012 TABLE 11 Cell uptake of compound 14 as compared to BPA 14 BPA Time Mean (ng/10.sup.6 cell) Time Mean (ng/10.sup.6 cell) (h) SAS B16F10 NIH-3T3 U87-MG (h) SAS B16F10 NIH-3T3 U87-MG 1.00 1 0 120 15 1.00 782 252 97.8 197 4.00 44 12 444 117 4.00 811 234 106 317 24.0 4 0 2 10 24 24.0 1345 267 75.9 172 indicates data missing or illegible when filed

TABLE-US-00013 TABLE 12 Cell uptake of compound 15 as compared to BPA 15 BPA Time Mean (ng/10.sup.6 cell) Time Mean (ng/10.sup.6 cell) (h) SAS B16F10 NIH-3T3 U87-MG (h) SAS B16F10 NIH-3T3 U87-MG 1.00 491 193 223 225 1.00 782 252 97.8 197 4.00 283 2 0 185 337 4.00 11 234 10 317 24.0 419 458 169 269 24.0 1345 267 75.9 172 indicates data missing or illegible when filed

TABLE-US-00014 TABLE 13 Cell uptake of compound 13 as compared to BPA 13 BPA Time Mean (ng/10.sup.6 cell) Time Mean (ng/10.sup.6 cell) (h) SAS B16F10 NIH-3T3 U87-MG (h) SAS B16F10 NIH-3T3 U87-MG 1 903 180 183 521 1 286 135 118 209 4 1012 310 222 828 4 225 261 112 247 24 1077 304 168 735 24 556 394 98.8 293

INCORPORATION BY REFERENCE

[0271] All of the U.S. patents and U.S. and PCT patent application publications cited herein are hereby incorporated by reference.

EQUIVALENTS

[0272] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.