Deuterated small molecule inhibitors of shared epitope-calreticulin interactions

Abstract

Selectively deuterated small molecules that modulate shared epitope-calreticulin (SE-CRT) binding have been synthesized. The pharmacokinetic properties of these new deuterated modulators of SE-CRT binding have also been measured. Methods of using the selectively deuterated small molecules are also described, including for the treatment of immunological disorders and inflammatory disorders.

Description

Attorney Reference: ALLN-001WODEUTERATED SMALL MOLECULE INHIBITORS OF SHARED EPITOPE- CALRETICULIN INTERACTIONSCROSS-REFERENCE TO RELATED APPLICATION

[0001] This application claims the benefit of the earlier filing date of United States Provisional Application Serial No. 63 / 788,714, filed April 14, 2025, the disclosure of which is incorporated herein by reference.INTRODUCTION

[0002] Rheumatoid arthritis (RA) and periodontal disease are both characterized by chronic inflammation and bone erosions.. Both diseases are associated with HLA-DRB1 alleles coding a five amino acid sequence motif called the ‘shared epitope’ (SE) in the region 70-74 of the DRp chain. In RA, the SE not only confers a higher risk, but also increases the likelihood of developing a more severe disease.

[0003] Calreticulin is a calcium-binding chaperone encoded in humans by the CALR gene. Calreticulin is thought to have one or more functions in immune regulation. It has been hypothesized that SE functions in part by activating calreticulin to induce its effects on bone destruction.

[0004] (R)-HL840 is a small molecule described as modulating shared epitope-calreticulin (SE-CRT) binding (Holoshitz et al., U. S. Patent 10,723,718). The chemical structure and systematic chemical name of (R)-HL840 is shown below:(R,E)-(1-((1-(3-(1-methyl-1H-pyrazol-4-yl)acryloyl)piperidin3-yl)methyl)-1H-1,2,3-triazol-4-yl)methanaminiumSUMMARY

[0005] Selectively deuterated small molecules that modulate shared epitope-calreticulin (SE-CRT) binding have been synthesized. The pharmacokinetic properties of these new deuterated modulators of SE-CRT binding have also been measured. Methods of using the selectivelyAttorney Reference: ALLN-001WOdeuterated small molecules are also described, including for the treatment of immunological disorders and inflammatory disorders.DETAILED DESCRIPTION

[0006] Selectively deuterated small molecules that modulate shared epitope-calreticulin (SE-CRT) binding have been synthesized. The pharmacokinetic properties of these new deuterated modulators of SE-CRT binding have also been measured. Methods of using the selectively deuterated small molecules are also described, including for the treatment of immunological disorders and inflammatory disorders.

[0007] Before the present invention is described in greater detail, it is to be understood that this invention is not limited to particular embodiments described, as such may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0008] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limits of that range is also specifically disclosed. Each smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in that stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range, and each range where either, neither or both limits are included in the smaller ranges is also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

[0009] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, some potential and exemplary methods and materials may now be described. Any and all publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. It is understood that the present disclosure supersedes any disclosure of an incorporated publication to the extent there is a contradiction.

[0010] It must be noted that as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. It is furtherAttorney Reference: ALLN-001WOnoted that the claims may be drafted to exclude any element, e.g., any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as “solely”, “only” and the like in connection with the recitation of claim elements, or the use of a “negative” limitation.

[0011] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed. To the extent the definition or usage of any term herein conflicts with a definition or usage of a term in an application or reference incorporated by reference herein, the instant application shall control.

[0012] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present invention. Any recited method can be carried out in the order of events recited or in any other order which is logically possible.DEFINITIONSIsotopes

[0013] “Hydrogen” refers to any isotope of hydrogen. However, the symbol “H” refers exclusively to the protium isotope (1H). In contrast, the symbol “D” refers to the deuterium isotope (2H) and the symbol “T” refers to the tritium isotope (3H).

[0014] Reference to any non-hydrogen atom by its name or chemical symbol includes all isotopes of that atom. For example, “carbon” or the symbol “C” includes12C,13C,14C, and all other isotopes of carbon. As another example, “oxygen” or the symbol “O” includes16O,17O,18O, and all other isotopes of oxygen.Organic chemistry

[0015] " Alkyl" refers to a monoradical, branched or linear, non-cyclic, saturated hydrocarbon group. Exemplary alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, octyl, decyl, cyclopentyl, and cyclohexyl. In some cases, the alkyl group has 1 to 24 carbon atoms, e.g. 1 to 12, 1 to 6, or 1 to 3. The term “Ci-Ce alkyl” refers to an alkyl group with from 1 to 6 carbon atoms.

[0016] “Alkenyl" refers to a monoradical, branched or linear, non-cyclic hydrocarbonyl group that comprises a carbon-carbon double bond. Exemplary alkenyl groups include ethenyl, n-Attorney Reference: ALLN-001WOpropenyl, isopropenyl, n-butenyl, isobutenyl, octenyl, decenyl, tetradecenyl, hexadecenyl, eicosenyl, and tetracosenyl.

[0017] “Alkynyl" refers to a monoradical, branched or linear, non-cyclic hydrocarbonyl group that comprises a carbon-carbon triple bond. Exemplary alkynyl groups include cthynyl and n-propynyl.

[0018] “Cycloalkyl” refers to a monoradical, cyclic, saturated hydrocarbon group. Similarly, “cycloalkenyl” refers to a monoradical and cyclic group having carbon-carbon double bond whereas “cycloalkynyl” refers to a monoradical and cyclic group having carbon-carbon triple bond.

[0019] “Cycloheteroalkyl” and “heterocyclyl” refers to a monoradical, cyclic group that contains a heteroatom (e.g. O, S, N) as a ring atom and that is not aromatic (i.e. distinguishing heterocyclyl groups from heteroaryl groups). Exemplary heterocyclyl groups include piperidinyl, tetrahydrofuranyl and dihydrofuranyl.

[0020] “Aryl" refers to an aromatic group containing at least one aromatic ring, wherein each of the atoms in the ring are carbon atoms, i.e. none of the ring atoms are heteroatoms (e.g. O, S, N). In some cases the aryl group has a second aromatic ring, e.g. that is fused to the first aromatic ring. Exemplary aryl groups are phenyl, naphthyl, biphenyl, diphenylether, diphenylamine, and benzophenone.

[0021] “Haloalkyl” refers to an alkyl group that has been substituted with at least one halogen atom, such as fluorine, chlorine, bromine, or iodine.

[0022] “Heteroaryl” refers to an aromatic group containing at least one aromatic ring, wherein at least one of the atoms in the aromatic ring is a heteroatom (e.g. O, S, N). Exemplary heteroaryl groups include those obtained from removing a hydrogen atom from pyridine, pyrimidine, furan, thiophene, or benzothiophene.

[0023] The term “heteroatom” includes the atoms of O, S, and N.

[0024] The tern “substituted” refers the removal of one or more hydrogens from an atom (e.g. from a C or N atom) and their replacement with a different group. For instance, a hydrogen atom on a phenyl (-CeHs) group can be replaced with a methyl group to form a -C6H4CH3 group. Thus, the -C6H4CH3 group can be considered a substituted aryl group. As another example, two hydrogen atoms from the second carbon of a propyl (-CH2CH2CH3) group can be replaced with an oxygen atom to form a -CH2C(O)CH3 group, which can be considered a substituted alkyl group. However, replacement of a hydrogen atom on a propyl (-CH2CH2CH3) group with a methyl group (e.g. giving -CH2CH(CH3)CH3) is not considered a “substitution” as used herein since the starting group and the ending group are both alkyl groups. However, if the propyl group was substituted with a methoxy group, thereby giving a -CH2CH(OCH3)CH3 group, theAttorney Reference: ALLN-001WOoverall group can no long be considered “alkyl”, and thus is “substituted alkyl”. Thus, in order to be considered a substituent, the replacement group is a different type than the original group. In addition, groups are presumed to be unsubstituted unless described as substituted. For instance, the term “alkyl” and “unsubstituted alkyl” are used interchangeably herein.

[0025] Exemplary substituents include alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof.

[0026] In some cases, the substitutions can themselves be further substituted with one or more groups. For example, the group -C6H4CH2CH3 can be considered as substituted aryl, i.e. an aryl group substituted with the ethyl, which is an alkyl group. Furthermore, the ethyl group can itself be substituted with a pyridyl group to form -C6H4CH2CH2C5H5N, wherein -C6H4CH2CH2C5H5N can also be considered as a substituted aryl group as the term is used herein. In some cases, the substituents are not substituted with any other groups.

[0027] Diradical groups are also described herein, i.e. in contrast to the monoradical groups such as alkyl and aryl described above. The term "alkylene" refers to the diradical version of an alkyl group, i.e. an alkylene group is a diradical, branched or linear, non-cyclic, saturated hydrocarbon group. Exemplary alkylene groups include diylmethane (-CH2-, which is also known as a methylene group), 1,2-diylethane (-CH2CH2-), and 1,1-diylethane (i.e. a CHCFE fragment where the first atom has two single bonds to other two different groups). The term “arylene” refers to the diradical version of an aryl group, e.g. 1,4-diylbenzene refers to a C.6H4 fragment wherein two hydrogens that are located para to one another are removed and replaced with single bonds to other groups. The terms “alkenylene”, “alkynylene”, “heteroarylene”, and “heterocyclene” are also used herein.

[0028] “Acyl” refers to a group of formula -C(O)R wherein R is alkyl, alkenyl, alkynyl, or substituted versions thereof. For example, the acetyl group has formula -C(O)CH3. “Carbonyl” refers to a diradical group of formula -C(O)-.

[0029] “Alkoxy" refers to a group of formula -O(alkyl). Similar groups can be derived from alkenyl, alkynyl, aryl, heteroaryl, and other groups.

[0030] “Amino" refers to the group -NRXRYwherein Rxand RYare each independently hydrogen or a non-hydrogen substituent. Exemplary non-hydrogen substituents include alkyl groups (e.g. methyl, ethyl, and isopropyl).

[0031] “Carbonyl” refers to a diradical group of formula -C(O)-.

[0032] “Carboxy” is used interchangeably with carboxyl and carboxylate to refer to the -CO2H group and salts thereof.Attorney Reference: ALLN-001WO

[0033] “Ether” refers to a diradical group of formula -O-. For instance, if the ether group is connected to an alkyl group, then the overall group is an alkoxy group (e.g. -OCH3 or methoxy). If the ether is connected to a carbonyl group, then the overall group is an ester group of formula -OC(O)-.

[0034] “Halo" and “halogen” refer to the chloro, bromo, fluoro, and iodo groups.

[0035] “Nitro” refers to the group of formula -NO2.Treatments

[0036] The terms active agent, active pharmaceutical ingredient, pharmacologically active agent, and drug arc used interchangeably herein to refer to a chemical material or compound which, when administered to an organism (human or animal) induces a desired pharmacologic and / or physiologic effect by local and / or systemic action.

[0037] The terms “individual,” “host,” “subject,” and “patient” are used interchangeably herein, and refer to an animal, including, but not limited to, human and non-human primates, including simians and humans; rodents, including rats and mice; bovines; equines; ovines; felines; canines; and the like. " Mammal" means a member or members of any mammalian species, and includes, by way of example, canines; felines; equines; bovines; ovines; rodentia, etc. and primates, e.g., non-human primates, and humans. Non-human animal models, e.g., mammals, e.g. non-human primates, murines, lagomorpha, etc. may be used for experimental investigations.

[0038] “Treating” or “treatment” as used herein concerns treatment of a disease or condition of interest in a patient or subject, particularly a human subject, having the disease or condition of interest, and includes by way of example, and without limitation:(i) inhibiting the disease or condition, for example, arresting or slowing its development;(ii) relieving the disease or condition, for example, causing regression of the disease or condition or a symptom thereof; or(iii) stabilizing the disease or condition.

[0039] “Preventing” as used herein concerns preventing the disease or condition from occurring in a patient or subject, in particular, when such patient or subject is predisposed to the condition but has not yet been diagnosed as having it.

[0040] A “therapeutically effective amount”, “effective amount”, a "therapeutically effective dose" or “therapeutic dose” is an amount sufficient to effect desired clinical results (i.e., achieve therapeutic efficacy, achieve a desired therapeutic response, etc.). A therapeutically effective dose can be administered in one or more administrations. For purposes of this disclosure, aAttorney Reference: ALLN-001WOtherapeutically effective dose of a compositions is an amount that is sufficient, when administered to the individual, to palliate, ameliorate, stabilize, reverse, prevent, slow or delay the progression of a disease state (e.g., cancer, etc.) present in the subject.

[0041] As used herein, the terms “determining,” “measuring,” “assessing,” and “assaying” are used interchangeably and include both quantitative and qualitative determinations.

[0042] The term “unit dosage form,” as used herein, refers to physically discrete units suitable as unitary dosages for human and animal subjects, each unit containing a predetermined quantity of a compound calculated in an amount sufficient to produce the desired effect in association with a pharmaceutically acceptable diluent, carrier or vehicle. The specifications for unit dosage forms depend on the particular compound employed and the effect to be achieved, and the pharmacodynamics associated with each compound in the host.

[0043] Also disclosed herein are salts of compounds of Formula (I). Such salts are formed between an acid and a basic group of the compound, such as an amino functional group, or a base and an acidic group of the compound, such as a carboxyl functional group. In certain embodiments, the present compounds are provided in the form of pharmaceutically acceptable acid addition salts.

[0044] The term “pharmaceutically acceptable,” as used herein, refers to a component that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and other mammals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit / risk ratio. A “pharmaceutically acceptable salt” means any non-toxic salt that, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of this invention. A “pharmaceutically acceptable counterion” is an ionic portion of a salt that is not toxic when released from the salt upon administration to a recipient.

[0045] Acids commonly employed to form pharmaceutically acceptable salts include inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid and phosphoric acid, as well as organic acids such as para-toluenesulfonic acid, salicylic acid, tartaric acid, bitartaric acid, ascorbic acid, maleic acid, besylic acid, fumaric acid, gluconic acid, glucuronic acid, formic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, lactic acid, oxalic acid, para-bromophenylsulfonic acid, carbonic acid, succinic acid, citric acid, benzoic acid and acetic acid, as well as related inorganic and organic acids. Such pharmaceutically acceptable salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate,Attorney Reference: ALLN-001WOsebacate, fumarate, maleate, butyne- 1,4-dioate, hexyne- 1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, sulfonate, xylene sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, f>-hydroxybutyratc, glycolatc, maleate, tartrate, mcthancsulfonatc, propancsulfonatc, naphthalcnc-1 -sulfonate, naphthalene-2-sulfonate, mandelate and other salts. In one embodiment, pharmaceutically acceptable acid addition salts of compounds of Formula (I) include those formed with mineral acids such as hydrochloric acid or hydrobromic acid, and especially those formed with organic acids such as maleic acid.

[0046] A "pharmaceutically acceptable excipient," "pharmaceutically acceptable diluent," "pharmaceutically acceptable carrier," and "pharmaceutically acceptable adjuvant" means an excipient, diluent, carrier, and adjuvant that are useful in preparing a pharmaceutical composition that are generally safe, non-toxic and neither biologically nor otherwise undesirable, and include an excipient, diluent, carrier, and adjuvant that are acceptable for veterinary use as well as human pharmaceutical use. " A pharmaceutically acceptable excipient, diluent, carrier and adjuvant" as used in the specification and claims includes both one and more than one such excipient, diluent, carrier, and adjuvant.

[0047] As used herein, a "pharmaceutical composition" is meant to encompass a composition suitable for administration to a subject, such as a mammal, especially a human. In general a “pharmaceutical composition” is sterile, and preferably free of contaminants that are capable of eliciting an undesirable response within the subject (e.g., the compound(s) in the pharmaceutical composition is pharmaceutical grade). Pharmaceutical compositions can be designed for administration to subjects or patients in need thereof via a number of different routes of administration including oral, buccal, rectal, parenteral, intraperitoneal, intradermal, intratracheal, intramuscular, subcutaneous, and the like.

[0048] The terms "co-administration" and "in combination with" include the administration of two or more therapeutic agents either simultaneously, concurrently or sequentially within no specific time limits. In one embodiment, the agents are present in the cell or in the subject's body at the same time or exert their biological or therapeutic effect at the same time. In one embodiment, the therapeutic agents are in the same composition or unit dosage form. In other embodiments, the therapeutic agents are in separate compositions or unit dosage forms. In certain embodiments, a first agent can be administered prior to (e.g., minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks before), concomitantly with, or subsequent to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24Attorney Reference: ALLN-001WOhours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks after) the administration of a second therapeutic agent.COMPOUNDS

[0049] Provided are compounds of Formula (I):or a pharmaceutically acceptable salt thereof,wherein:the compound is enriched in deuterium;R1is -[C(R11)2]n-NRaRa;R2is selected from H, Rb, and Rbsubstituted with one or more C?-Cs cycloalkyl; each R11is independently selected from hydrogen and Ci-Ce alkyl;n is 1, 2, or 3;each Rais independently selected from hydrogen, Ci-Ce alkyl, Ci-Ce haloalkyl, and -C(O)Rb, or two Raare taken together with the nitrogen atom to which they are bonded to form a 5- to 8-membered cycloheteroalkyl which may optionally include one or more of the same or different additional heteroatoms and which may optionally be substituted with one or more of the same or different Rb, -C(O)Rb, or both; andeach Rbis independently selected from Ci-Ce alkyl, -(Ci-Ce alkylcncXCj-Cs cycloalkyl), Ci-C6haloalkyl, -(Ci-Ce haloalkyl)(Cj-Cs cycloalkyl), and Cj-Cs cycloalkylCompounds and isotopic enrichment

[0050] The tern “compound” as used herein refers to a collection of molecules having an identical chemical structure except for one or more isotopic variations among the constituent atoms of the molecules. For example, if two molecules are identical except that one has carbon-12 at position 8 whereas the other molecule has carbon- 13 at position 8, then both molecules are the same compound. As another example, if two molecules are identical except that one is -12CH2- at position 10 whereas the other molecule is -12CD2- at position 10, then both molecules are the same compound. The term “isotopologue” refers to compounds that have identical structures except for one or more isotopic variations.Attorney Reference: ALLN-001WO

[0051] The compounds described herein are “enriched in deuterium’’, which means that at least one chemical position labeled as hydrogen in the compound has a proportion of deuterium (D) that is significantly greater than the natural isotopic abundance of deuterium. For example, the natural isotopic abundance of deuterium (D) is about 0.0145%, whereas almost all other naturally occurring hydrogen atoms are protium (H). Therefore, if the proportion of deuterium at a specific chemical position in the compound is significantly above 0.0145%, then the compound is enriched in deuterium at the specific chemical position. Hence, the compound is also “enriched in deuterium” in general.

[0052] As used herein, the term “proportion of deuterium” refers to mole ratios and does not refer to mass ratios. The term “deuteration position” refers to a position in a compound that is enriched in deuterium.

[0053] The term “isotopic enrichment factor” in relation to deuterium is the ratio between the actual proportion of deuterium at a specific chemical position and the natural isotopic abundance of deuterium. For example, a deuterium isotopic enrichment factor of 100 means that 1.45% of the chemical position is deuterium (D), e.g. and about 98.55% is protium (H). A deuterium isotopic enrichment factor of 1,000 means that 14.5% of the chemical position is deuterium, e.g. and about 85.5% is protium.

[0054] The mole proportion of deuterium at a particular chemical position can also be quantified as a percentage of “deuterium incorporation”. If 14.5 mol% of a particular chemical position is deuterium, then the particular chemical position can also be described as having 14.5% deuterium incorporation and a deuterium isotopic enrichment factor of 1,000.

[0055] In some cases, the deuterium incorporation at a particular chemical position is 10% or more, such as 25% or more, 50% or more, 75% or more, 80% or more, 85% or more, 90% or more, 95% or more, 98% or more, or 99% or more. In some cases, the deuterium isotopic enrichment factor at a particular chemical position is 100 or more, such as 1,000 or more, 3,000 or more, 5,000 or more, 6,000 or more, 6,500 or more, or 6,800 or more.

[0056] In some embodiments, the compound is enriched in deuterium at one or more chemical positions that each have deuterium incorporation of 50% or more, such as 80% or more.

[0057] In some embodiments, the compound is enriched in deuterium at two or more chemical positions that each have deuterium incorporation of 50% or more, such as 80% or more.

[0058] In some embodiments, the compound is enriched in deuterium at four or more chemical positions that each have deuterium incorporation of 50% or more, such as 80% or more.Attorney Reference: ALLN-001WO

[0059] In some cases, the position of deuterium isotopic enrichment includes two or more hydrogen atoms. For example, position 10 in Formula (I) includes a carbon atom bonded to two hydrogen atoms, one nitrogen atom, and one carbon atom. In cases where a position includes two or more hydrogen atoms, overall deuterium incoiporation is the arithmetic mean of deuterium incorporation at each hydrogen position. For example, if 60% of molecules have two deuterium atoms, 30% of molecules have one deuterium and one protium, and 10% of molecules have two protium atoms, then deuterium incorporation is 0.6(100%) + 0.3(50%) + 0.1(0%) = 75%.

[0060] In some cases, the enrichment in deuterium is selective, i.e. the deuterium incorporation at two or more particular chemical positions is significantly different. For example, the compound can have two chemical positions that have deuterium proportions that differ by 10-fold or more, such as 25-fold or more, 100-fold or more, or 1000-fold or more. For example, a deuteration position can have deuterium incoiporation of 90% whereas a protiation position can have only 0.09% deuterium, which is a 1000-fold difference.

[0061] Similarly, in some cases positions 1, 2, 11, or a combination thereof are each -CH=. In some cases, position 6, position 7, or a combination thereof are each -CH2-. In such cases, the positions are not significantly enriched in deuterium. For example, the amount of hydrogen atoms at such positions that are H (i.e. protium) can be 99% or more, such as 99.9% or more. Structural positions in Formula (I)

[0062] For the purpose of reference, positions in the skeletal structure of Formula (I) are illustrated below:wherein each numbered position may be deuterated.

[0063] As stated above, R1is -[C(R11)2]n-NRaRa. In some embodiments, each R11is Y, wherein each Y is independently H or D. Additionally, since n is 1 or 2 or 3, then the left-hand half of the R1group can be -CY2-, -CY2CY2-, or -CY2CY2CY2-. In other words, the left half of R1can be a methylene, ethylene, or propylene linkage. In some cases, the left half of R1is a methylene linkage, such as -CH2-, -CHD- or -CD2-.

[0064] The right half of R1is NRaRa. For example, the first Ragroup can be Y. As stated above, each Y is independently selected from H (protium) and D (deuterium). In some cases,Attorney Reference: ALLN-001WOthe second Rais also Y, which can be H or D. Therefore, in some cases, the right-hand half of R1is NY2, such as NH2.

[0065] In some embodiments, the first Rais Y but the second Rais a non-Y group. For example, the first Racan be Y and the second Racan be Ci-Ce alkyl or -C(O)Rb. For example, the second Racan be -C(O)(methyl), such as -C(O)(CHa).

[0066] The various aspects of R1can be chosen in any suitable combination. The different aspects of R1are: the value of n, the identity of each R11group, the first Ra, and the second Ra. In some cases, R1is -[CY2]n-NRaRa. In some cases, R1is -CY2-NRaRa. In some cases, each Rais independently selected from Y, Ci-Ce alkyl, and -C(O)Rb, wherein Rbis Ci-Ce alkyl. In some cases, R1is -[CY2]n-NH2 or -[CY2]n-NH-C(O)(Rb). In some cases, R1is -[CY2]n-NH2 or -[CY2]n-NY-C(O)(methyl). In some cases, R1is -CD2NH2 or -CD2NC(O)(CH3). In some cases, R1is -CH2NH2 or -CH2NC(O)(CH3).

[0067] In some embodiments, R1is enriched in deuterium, i.e. one or more position within R1are enriched in deuterium. For example, [C(R11)2]n can be -CD2-.

[0068] As stated above, R2is selected from H, Rb, and Rbsubstituted with one or more Ca-Cs cycloalkyl. In some cases, R2is an Rbgroup of Ci-Ce alkyl or C1-C6 haloalkyl. In some cases, R2is an Rbgroup of methyl. In some cases, R2is -CH3 (i.e. protiated methyl) or -CD3 (i.e. deuterated methyl). In some embodiments, R2is the Rbgroup of-(Ci-Ce alkylene)(C3-Cs cycloalkyl), such as -(methylene)(cyclopropyl). In some embodiments, R2is enriched in deuterium, e.g. R2is -CD3.

[0069] Formula (I) shows labels for positions 1 through 11. In some cases, positions 3 and 4 are each enriched in deuterium. In some cases, positions 3 and 4 are each -CD=. Structures herein depicting D at a particular position indicate that each D represents deuterium incorporation of 50% or more, such as 70%, 80%, 90%, or more. Working examples had greater than 90% deuterium incorporation at each position. Therefore, stating that position 3 is -CD= means that 50% or more of the molecules in the compound have carbon bonded to a deuterium atom at position 3, and therefore 50% or less of the molecules have carbon bonded to a protium atom at position 3.

[0070] In some cases, positions 5 and 9 are each enriched in deuterium. In some cases, positions 5 and 9 are each -CD2-. In some cases, position 8 is enriched in deuterium. In some cases, position 8 isAttorney Reference: ALLN-001WO

[0071] In some cases, position 10 is enriched in deuterium. In some cases, position 10 is -CD2-.

[0072] In some cases, each D represent deuterium incorporation of 60% or more, such as 70% or more, 80% or more, 90% or more, 95% or more, or 98% or more.

[0073] In some cases, positions 1, 2, 11, or a combination thereof are -CH=. In some cases, the proportion of protium (H) at position 1, 2, 11, or a combination thereof is 60% or more, 70% or more, 80% or more, 90% or more, 95% or more, 99% or more, or 99.9% or more. Similarly, in some cases, position 6, position 7, or a combination thereof are each -CH2-. In some cases, the proportion of protium (H) at position 6, 7, or a combination thereof is 60% or more, 70% or more, 80% or more, 90% or more, 95% or more, 99% or more, or 99.9% or more.

[0074] Formula (I) shows the carbon-carbon double bond at positions 3 and 4 as having an ambiguous cis / trans configuration. In some cases, the carbon-carbon double bond at positions 3 and 4 has a trans configuration.PHARMACEUTICAL COMPOSITIONS

[0075] Provided are pharmaceutical compositions that include a compound as described herein, such as a compound according to Formula (I). Such pharmaceutical compositions may be manufactured by any suitable method, such as mixing, dissolving, granulating, drageemaking, levigating, emulsifying, encapsulating, entrapping or lyophilization processes. The compositions may be formulated using one or more physiologically acceptable excipients, diluents, carriers, adjuvants or auxiliaries to provide preparations which can be used pharmaceutically.

[0076] The active compound(s) may be formulated in the pharmaceutical compositions per se, or in the form of a hydrate, solvate, or pharmaceutically acceptable salt. Typically, such salts are more soluble in aqueous solutions than the corresponding free acids and bases, but salts having lower solubility than the corresponding free acids and bases may also be formed.

[0077] Pharmaceutical compositions comprising the disclosed compound(s) may take a form suitable for virtually any mode of administration, including, for example, topical, ocular, oral, buccal, systemic, nasal, injection, such as i.v. or i.p., transdermal, rectal, vaginal, etc., or a form suitable for administration by inhalation or insufflation.

[0078] For topical administration, the active compound(s) (or a hydrate, solvate, N-oxide or pharmaceutically acceptable salt thereof) may be formulated as solutions, gels, ointments, creams, suspensions, etc. as are well-known in the art.Attorney Reference: ALLN-001WO

[0079] Systemic formulations include those designed for administration by injection, e.g., subcutaneous, intravenous, intramuscular, intrathecal or intraperitoneal injection, as well as those designed for transdermal, transmucosal oral or pulmonary administration.

[0080] Useful injectable preparations include sterile suspensions, solutions, or emulsions of the active compound(s) in aqueous or oily vehicles. The compositions may also contain formulating agents, such as suspending, stabilizing and / or dispersing agent. The formulations for injection may be presented in unit dosage form, e.g., in ampules or in multidose containers, and may contain added preservatives.

[0081] Alternatively, the injectable formulation may be provided in powder form for reconstitution with a suitable vehicle, including but not limited to sterile, pyrogen-free water, buffer, dextrose solution, etc., before use. To this end, the active compound(s) may be dried by any art-known technique, such as lyophilization, and reconstituted prior to use.

[0082] For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are known in the art.

[0083] For oral administration, the pharmaceutical compositions may take the form of, for example, lozenges, tablets or capsules prepared by conventional means with pharmaceutically acceptable excipients, such as: binding agents (e.g., pregelatinized maize starch, polyvinylpyrrolidone or hydroxypropyl methylcellulose); fillers (e.g., lactose, microcrystalline cellulose or calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc or silica); disintegrants (e.g., potato starch or sodium starch glycolate); and / or wetting agents (e.g., sodium lauryl sulfate). The tablets may be coated by methods well known in the art with, for example, sugars, films or enteric coatings.

[0084] Liquid preparations for oral administration may take the form of, for example, elixirs, solutions, syrups or suspensions, or they may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations may be prepared by conventional means with pharmaceutically acceptable excipients such as: suspending agents (e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifying agents (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol, cremophoreTM. or fractionated vegetable oils); and preservatives (e.g., methyl or propyl-p-hydroxybenzoates or sorbic acid). The preparations may also contain buffer salts, preservatives, flavoring, coloring and sweetening agents as appropriate.

[0085] Preparations for oral administration may be suitably formulated to give controlled release of the active compound, as is well known.

[0086] For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.Attorney Reference: ALLN-001WO

[0087] For rectal and vaginal routes of administration, the active compound(s) may be formulated as solutions (for retention enemas) suppositories or ointments containing conventional suppository bases, such as cocoa butter or other glycerides.

[0088] For nasal administration or administration by inhalation or insufflation, the active compound(s), hydrate, solvate, N-oxide, or pharmaceutically acceptable salt can be conveniently delivered in the form of an aerosol spray from pressurized packs or a nebulizer with the use of a suitable propellant, e.g.,) dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, fluorocarbons, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges for use in an inhaler or insufflator (for example capsules and cartridges comprised of gelatin) may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.

[0089] The pharmaceutical compositions can be in the form of a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution and isotonic sodium chloride solution.

[0090] For ocular administration, the active compound(s) may be formulated as a solution, emulsion, suspension, etc. suitable for administration to the eye. A variety of vehicles suitable for administering compounds to the eye are known in the art. Specific non-limiting examples are described in U. S. Pat. Nos. 6,261,547; 6,197,934; 6,056,950; 5,800,807; 5,776,445;5,698,219; 5,521,222; 5,403,841; 5,077,033; 4,882,150; and 4,738,851, which are incorporated herein by reference.

[0091] For prolonged delivery, the active compound(s) can be formulated as a depot preparation for administration by implantation or intramuscular injection. The active ingredient may be formulated with suitable polymeric or hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, e.g., as a sparingly soluble salt. Alternatively, transdermal delivery systems manufactured as an adhesive disc or patch which slowly releases the active compound(s) for percutaneous absorption may be used. To this end, permeation enhancers may be used to facilitate transdermal penetration of the active compound(s). Suitable transdermal patches are described in for example, U. S. Pat. Nos.5,407,713; 5,352,456; 5,332,213; 5,336,168; 5,290,561; 5,254,346; 5,164,189; 5,163,899;5,088,977; 5,087,240; 5,008,110; and 4,921,475, which are incoiporated herein by reference.Attorney Reference: ALLN-001WO

[0092] Alternatively, other pharmaceutical delivery systems may be employed. Liposomes and emulsions are well-known examples of delivery vehicles that may be used to deliver active compound(s). Certain organic solvents, such as dimethylsulfoxide (DMSO), may also be employed, although usually at the cost of greater toxicity.

[0093] The pharmaceutical compositions may, if desired, be presented in a pack or dispenser device which may contain one or more unit dosage forms containing the active compound(s). The pack may, for example, comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration.

[0094] The disclosed compounds, including those of Formula (I) may be used alone, in any combination, and in combination with, or adjunctive to, at least one second therapeutic agent. Furthermore, the disclosed compound or compounds, and / or the at least one second therapeutic, may be used in combination with any suitable excipient useful for forming compositions for administration to a subject. Excipients can be included in pharmaceutical compositions for a variety of purposes, such as to dilute a composition for delivery to a subject, to facilitate processing of the formulation, to provide advantageous material properties to the formulation, to facilitate dispersion from a delivery device, to stabilize the formulation (e.g., antioxidants or buffers), to provide a pleasant or palatable taste or consistency to the formulation, or the like. The pharmaceutically acceptable excipient(s) may include a pharmaceutically acceptable carrier(s) and / or a pharmaceutically acceptable adjuvant(s). Exemplary excipients include, but are not limited to: mono-, di-, and polysaccharides, sugar alcohols and other polyols, such as, lactose, glucose, raffinose, melezitose, lactitol, maltitol, trehalose, sucrose, mannitol, starch, or combinations thereof; surfactants, such as sorbitols, diphosphatidyl choline, and lecithin; bulking agents; buffers, such as phosphate and citrate buffers; anti-adherents, such as magnesium stearate; binders, such as saccharides (including disaccharides, such as sucrose and lactose,), polysaccharides (such as starches, cellulose, microcrystalline cellulose, cellulose ethers (such as hydroxypropyl cellulose)), gelatin, synthetic polymers (such as polyvinylpyrrolidone, polyalkylene glycols); coatings (such as cellulose ethers, including hydroxypropylmethyl cellulose, shellac, com protein zein, and gelatin); release aids (such as enteric coatings); disintegrants (such as crospovidone, crosslinked sodium carboxymethyl cellulose, and sodium starch glycolate); fillers (such as dibasic calcium phosphate, vegetable fats and oils, lactose, sucrose, glucose, mannitol, sorbitol, calcium carbonate, and magnesium stearate); flavors and sweeteners (such as mint, cherry, anise, peach, apricot or licorice, raspberry, and vanilla); lubricants (such as minerals, exemplified by talc or silica, fats, exemplified by vegetable stearin, magnesium stearate or stearic acid); preservatives (such as antioxidants exemplified by vitamin A, vitamin E, vitamin C, retinyl palmitate, and selenium, amino acids, exemplified by cysteineAttorney Reference: ALLN-001WOand methionine, citric acid and sodium citrate, parabens, exemplified by methyl paraben and propyl paraben); colorants; compression aids; emulsifying agents; encapsulation agents; gums; granulation agents; and combinations thereof.METHODS OF INHIBITING

[0095] Provided are methods of inhibiting shared epitope (SE) - calreticulin (CRT) binding that include contacting the calreticulin with an effective amount of a compound as described herein.

[0096] In some cases, the contacting occurs in vitro. In other cases, the contacting occurs in vivo. In some cases, the calreticulin is in an osteoclast, e.g. wherein the contacting inhibits osteoclastogenesis. In some cases, the contacting includes administering the compound as described herein to a subject.METHODS OF USE

[0097] Provided are methods for using the disclosed compounds. For example, in a method of treating a patient for a condition that include administering an effective amount of a compound described herein to the patient. Also described herein is the use of the present compounds in the manufacture of a medicament for treating a condition.

[0098] In some cases, the condition is mediated by shared epitope (SE) - calreticulin (CRT) binding. In some cases, the condition is an immunological disorder or an inflammatory disorder.

[0099] The disclosed compound or combinations of disclosed compounds will generally be used in an amount effective to achieve the intended result, for example, in an amount effective to treat, prevent or ameliorate a particular condition. The disclosed compound(s), or compositions thereof, can be administered therapeutically to achieve therapeutic benefit or prophylactically to achieve a prophylactic benefit. Therapeutic benefit means eradication or amelioration of the underlying disorder being treated and / or eradication or amelioration of one or more of the symptoms associated with the underlying disorder such that the patient reports an improvement in feeling or condition, notwithstanding that the patient may still be afflicted with the underlying disorder. For example, administration of a compound to a patient suffering from an allergy provides therapeutic benefit not only when the underlying allergic response is eradicated or ameliorated, but also when the patient reports a decrease in the severity or duration of the symptoms associated with the allergy following exposure to the allergen. As another example, therapeutic benefit in the context of asthma includes an improvement in respiration following the onset of an asthmatic attack or a reduction in the frequency or severity of asthmatic episodes.Attorney Reference: ALLN-001WOTherapeutic benefit also includes halting or slowing the progression of the disease, regardless of whether improvement is realized.

[0100] As known by those of ordinary skill in the art, the preferred dosage of disclosed compounds may depend on various factors, including the age, weight, general health, and severity of the condition of the patient or subject being treated. Dosage also may need to be tailored to the sex of the individual and / or the lung capacity of the individual, when administered by inhalation. Dosage may also be tailored to individuals suffering from more than one condition or those individuals who have additional conditions that affect lung capacity and the ability to breathe normally, for example, emphysema, bronchitis, pneumonia, and respiratory infections. Dosage, and frequency of administration of the disclosed compound(s) or compositions thereof, will also depend on whether the disclosed compound(s) are formulated for treatment of acute episodes of a condition or for the prophylactic treatment of a disorder. A person of ordinary skill in the art will be able to determine the optimal dose for a particular individual.

[0101] For prophylactic administration, the disclosed compound, combinations of disclosed compounds, or compositions thereof, can be administered to a patient or subject at risk of developing one of the previously described conditions. For example, if it is unknown whether a patient or subject is allergic to a particular drug, the disclosed compound, combinations of disclosed compounds, or compositions thereof, can be administered prior to administration of the drug to avoid or ameliorate an allergic response to the drug. Alternatively, prophylactic administration can be used to avoid or ameliorate the onset of symptoms in a patient diagnosed with the underlying disorder. For example, a disclosed compound(s), or composition thereof, can be administered to an allergy sufferer prior to expected exposure to the allergen. A disclosed compound, combinations of disclosed compounds, or compositions thereof, can also be administered prophylactically to healthy individuals who are repeatedly exposed to agents known to one of the above-described maladies to prevent the onset of the disorder. For example, a disclosed compound, combinations of disclosed compounds, or compositions thereof, can be administered to a healthy individual who is repeatedly exposed to an allergen known to induce allergies, such as latex, in an effort to prevent the individual from developing an allergy. Alternatively, a disclosed compound, combinations of disclosed compounds, or compositions thereof, can be administered to a patient suffering from asthma prior to partaking in activities which trigger asthma attacks to lessen the severity of, or avoid altogether, an asthmatic episode.

[0102] Effective dosages can be estimated initially from in vitro assays. For example, an initial dosage for use in subjects can be formulated to achieve a circulating blood or serumAttorney Reference: ALLN-001WOconcentration of active compound that is at or above an IC50 or EC50 of the particular compound as measured in an in vitro assay. Dosages can be calculated to achieve such circulating blood or serum concentrations, taking into account the bioavailability of the particular compound. See, Goodman and Gilman’s The Pharmacological Basis of Therapeutics, 14th cd. McGraw-Hill Education, 2022, and the references cited therein, provide additional guidance concerning effective dosages.

[0103] Initial dosages can also be estimated from in vivo data, such as animal models.Animal models useful for testing the efficacy of compounds to treat or prevent the various diseases described above are well-known in the art. Dosage amount and dosage interval can be adjusted for individuals to provide plasma levels of the disclosed compound that are sufficient to maintain therapeutic or prophylactic effect. For example, the compounds can be administered once per day, multiple times per day, once per week, multiple times per week (e.g., every other day), one per month, multiple times per month, or once per year, depending upon, amongst other things, the mode of administration, the specific indication being treated, and the judgment of the prescribing physician. Persons of ordinary skill in the art will be able to optimize effective local dosages without undue experimentation.

[0104] In some cases, the condition is an immunoregulatory abnormality, e.g. selected from the group consisting of: systemic lupus erythematosis, chronic rheumatoid arthritis, type I diabetes mellitus, type II diabetes, inflammatory bowel disease, biliary cirrhosis, multiple sclerosis (MS), Crohn's disease, ulcerative colitis, bullous pemphigoid, sarcoidosis, psoriasis, autoimmune myositis, Wegener’s granulomatosis, ichthyosis, Graves ophthalmopathy, asthma, transplantation of organs or tissue, graft - versus - host diseases brought about by transplantation, autoimmune syndromes including rheumatoid arthritis, systemic lupus erythematosus, Hashimoto’s thyroiditis, myasthenia gravis, uveitis, posterior uveitis, allergic encephalomyelitis, glomerulonephritis, post-infectious autoimmune dis eases including rheumatic fever and post-infectious glomerulonephritis, inflammatory and hyperproliferative skin diseases, psoriasis, atopic dermatitis, contact dermatitis, eczematous dermatitis, seborrhoeic dermatitis, lichen planus, pemphigus, bullous pemphigoid, epidermolysis bullosa, urticaria, angioedemas, vasculitis, erythema, cutaneous eosinophilia, lupus erythematosus, acne, alopecia areata, keratoconjunctivitis, vernal conjunctivitis, uveitis associated with Behcet’s disease, keratitis, herpetic keratitis, conical cornea, dystrophia epithelialis comeae, corneal leukoma, ocular pemphigus, Mooren’s ulcer, scleritis, Graves’ opthalmopathy, Vogt-Koyanagi-Harada syndrome, sarcoidosis, pollen allergies, reversible obstructive airway disease, bronchial asthma, allergic asthma, intrinsic asthma, extrinsic asthma, dust asthma, chronic or inveterate asthma, late asthma and airway hyper-responsiveness, bronchitis, gastric ulcers, vascular damage causedAttorney Reference: ALLN-001WOby ischemic diseases and thrombosis, ischemic bowel diseases, inflammatory bowel diseases (IBD), necrotizing enterocolitis, intestinal lesions associated with thermal bums, coeliac diseases, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn’s disease, ulcerative colitis, migraine, rhinitis, eczema, interstitial nephritis, Goodpasture’s syndrome, hemolytic-uremic syndrome, diabetic nephropathy, multiple myositis, Guillain-Barre syndrome, Meniere’s disease, polyneuritis, multiple neuritis, mononeuritis, radiculopathy, hyperthyroidism, Basedow’s disease, pure red cell aplasia, aplastic anemia, hypoplastic anemia, idiopathic thrombocytopenic puipura, autoimmune hemolytic anemia, agranulocytosis, pernicious anemia, megaloblastic anemia, anerythroplasia, osteoporosis, sarcoidosis, fibroid lung, idiopathic interstitial pneumonia, dermatomyositis, leukoderma vulgaris, ichthyosis vulgaris, photoallergic sensitivity, cutaneous T cell lymphoma, arteriosclerosis, atherosclerosis, aortitis syndrome, polyarteritis nodosa, myocardosis, scleroderma, Wegener’s granuloma, Sjogren’s syndrome, adiposis, eosinophilic fascitis, lesions of gingiva, periodontium, alveolar bone, substantia osseadentis, glomerulonephritis, male pattern alopecia or alopecia senilis by preventing epilation or providing hair germination and I or promoting hair generation and hair growth, muscular dystrophy, pyoderma and Sezary’s syndrome, Addison’s disease, ischemia - reperfusion injury of organs which occurs upon preservation, transplantation or ischemic disease, endotoxin -shock, pseudomembranous colitis, colitis caused by drug or radiation, ischemic acute renal insufficiency, chronic renal insufficiency, toxinosis caused by lung - oxygen or drugs, lung cancer, pulmonary emphysema, cataracta, siderosis, retinitis pigmentosa, senile macular degeneration, vitreal scarring, corneal alkali burn, dermatitis erythema multiforme, linear IgA ballons dermatitis and cement dermatitis, gingivitis, periodontitis, sepsis, pancreatitis, dis eases caused by environmental pollution, aging, carcinogenesis, metastasis of carcinoma and hypobaropathy, disease caused by histamine or leukotriene release, Behcet’s disease, autoimmune hepatitis, primary biliary cirrhosis, sclerosing cholangitis, partial liver resection, acute liver necrosis, necrosis caused by toxin, viral hepatitis, shock, or anoxia, B-virus hepatitis, non-A / non-B hepatitis, cirrhosis, alcoholic cirrhosis, hepatic failure, fulminant hepatic failure, late - onset hepatic failure, "acute-on-chronic ” liver failure, augmentation of chemotherapeutic effect, cytomegalovirus infection, human cytomegalovirus (HCMV) infection, acquired immune deficiency syndrome (AIDS), cancer, senile dementia, trauma, chronic bacterial infection, rejection of transplanted organ or tissue, insulin dependent diabetes, and non-insulin dependent diabetes.

[0105] In some cases, the condition is selected from the group consisting of rheumatoid arthritis, multiple sclerosis (MS), systemic lupus erythematosus, psoriasis, rejection ofAttorney Reference: ALLN-001WOtransplanted organ or tissue, inflammatory bowel disease (IBD), insulin dependent diabetes, non-insulin dependent diabetes, and periodontitis.

[0106] In some cases, the condition includes bone erosion, bone loss, or a combination thereof.

[0107] In some cases, the condition is a neuroinflanmiatory disorder. In particular, the present compounds are useful for treating or preventing neuroinflammatory conditions.Examples of such conditions include, without limitation, Alzheimer’s disease, multiple sclerosis, Parkinson’s disease and the like.

[0108] In some cases, the condition treated with one or more of the present compounds, such as a compound of Formula (I), includes a hematopoietic neoplasm, such as a myeloproliferative disorder (MPN). In some cases, the hematopoietic neoplasm is selected from leukemias and lymphomas.

[0109] In some cases, the condition is selected from the group consisting of rheumatoid arthritis, multiple sclerosis (MS), systemic lupus erythematosus, psoriasis, rejection of transplanted organ or tissue, inflammatory bowel disease (IBD), uveitis, Parkinson’s disease, insulin dependent diabetes, and non-insulin dependent diabetes.

[0110] In some cases, the method further includes one or more anti-inflammatory compounds, one or more anti-rheumatic compounds, or a combination thereof to the patient.EXAMPLES

[0111] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention nor are they intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g. amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric.Overview

[0112] Examples 1 - 22 describe the synthesis of the selectively-deuterated compounds shown below. Such compounds were also characterized by a battery of analytical methods including liquid chromatography-mass spectroscopy (LC-MS), proton NMR (1H NMR, and high performance liquid chromatography (HPLC). The deuterium enrichment level was calculated with the IsoPat2software program based on mass spectroscopy data. Example 23 describes theAttorney Reference: ALLN-001WOassessment of the pharmacokinetic properties of the synthesized compounds in rodents.Example 24 provides an assessment of pharmacokinetic properties of exemplary compounds in dogs.Attorney Reference: ALLN-001WO

[0113] In some embodiments, a compound according Formula (I), or a salt thereof, has at least one improved property compared to known modulators of shared epitope (SE) -calreticulin (CRT) binding. The one or more improved property may include, but are not limited to, physical properties, including but not limited to, melting point, glass transition temperature, flowability, and / or stability, such as thermal stability, mechanical stability, shelf life, stability against polymorphic transition, and the like; chemical properties, such as, but not limited to, hygroscopic properties, solubility in water and / or organic solvents, reactivity, compatibility with excipients and / or delivery vehicles; and / or pharmacokinetic and / or pharmacodynamic properties, such as, but not limited to, bioavailability, absorption, distribution, metabolism, excretion, toxicity including cytotoxicity, dissolution rate, selectivity, activity, and / or half-life. In one aspect, selectivity includes on-target versus off-target selectivity, e.g. inhibition of SE -CRT binding. In other aspects, selectivity includes selectivity for one tissue in a subject, asAttorney Reference: ALLN-001WOindicated for example by a partition coefficient. In certain embodiments, the present compounds exhibit at least one improved property relative to a compound disclosed in U. S. Patent No.10,723,718.Deuteration strategy

[0114] As described in the Examples below, deuterium atoms were installed in various locations of the molecules through various techniques.

[0115] Methyl-d3 groups were installed at the R2position of Formula (I) by performing a nucleophilic attack on CD3I (see Example 1). Two geminal deuterium atoms were installed at carbon-8 by reducing a carboxylate group with NaBD4 (see Example 2). Hydrogen atoms at carbons 3 and 7 were exchanged for deuteriums with a base-catalyzed reflux reaction usingNaOD in D2O (see Example 3). One deuterium was installed at each of carbons 1 and 2 by reducing an alkyne into an alkene in the presence of D2O (see Example 4). Two deuteriums were installed on the R1group by reducing an ester with NaBD4 (see Example 6). A deuterium was installed at tertiary carbon 6 by refluxing with NaOD and D2O in a base-catalyzed reaction that exchanged a hydrogen atom for a deuterium atom (see Example 19).Example 1: Preparation of R, E)- l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-oneStep 1:

[0116] To a solution of benzyl (R)-3-(hydroxymethyl)piperidine-l-carboxylate la (15 g, 60.166 mmol) in toluene (80 mL) was added DPPA (23.18 g, 84.232 mmol) at room temperature under nitrogen atmosphere, followed by the addition of DBU (12.82 g, 84.232 mmol). The resulting mixture was heated at 110°C for 2 h. The reaction progress was monitored by LCMS.Attorney Reference: ALLN-001WOAfter cooling to room temperature, the reaction mixture was concentrated under reduced pressure. The residue was dissolved in water (100 mL). The aqueous mixture was extracted with EA (3 x 100 mL). The combined organic extracts were washed with brine (2 x 100 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CI LCL / McOI I (9:1), to give benzyl (R)-3-(azidomethyl)piperidine-l -carboxylate lb (12 g, 72.71%) as a yellow oil. LC-MS (ESI): m / z, [M+H]+= 275.15.Step 2:

[0117] To a stirred solution of benzyl (R)-3-(azidomethyl)piperidine- 1 -carboxylate lb (10 g, 36.453 mmol) in t-BuOH (50 mL) and H2O (30 mL) was added tert-butyl N-(prop-2-yn-l-yl)carbamate (7.35 g, 47.389 mmol) at room temperature, followed by the addition of CuSO4.5H2O (1.82 g, 7.291 mmol). To the above mixture was added sodium ascorbate (1.44 g, 7.291 mmol). The resulting mixture was stirred at room temperature for 2 h, and concentrated under reduced pressure. The residue was dissolved in water (50 mL). The aqueous mixture was extracted with EA (3 x 100 mL). The combined organic extracts were washed with brine (2 x 100 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to a residue, which was purified by reversed-phase flash column chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 0% to 10% gradient in 10 min; detector, UV 254 nm. Removal of solvents afforded benzyl (R)-3-(4-(((tert-butoxycarbonyl)amino)methyl)-lH-l, 2, 3-triazoLl -yl)methyl)piperidine-l -carboxylate 1c (13.2 g, 84.31%) as a white solid. LC-MS (ESI): m / z [M+H]+= 430.25.Step 3:

[0118] To a stirred solution of benzyl (R)-3-((4-(((tert-butoxycarbonyl)amino)methyl)-lH-1, 2, 3-triazol-l-yl)methyl)piperidine-l -carboxylate 1c (13 g, 30.266 mmol) in methanol (130 mL) was added Pd / C (6.44 g) at room temperature. The resulting mixture was stirred and heated at 50°C for 16 h under a hydrogen atmosphere. The reaction progress was monitored by LCMS. The reaction mixture was filtered, the filter cake was washed with MeOH (3 x 100 mL). The combined filtrates were concentrated under reduced pressure to provide tert-butyl (R)-(1-(piperidin-3-ylmethyl)-1H-1,2,3-triazol-4-yl)methyl)carbamate 1d (7.8 g, 87.25%) as a light yellow semi-solid. This product was used in the next step directly without further purification. LC-MS (ESI): m / z [M+H]+= 296.25.Step 4:

[0119] To a stirred solution of lH-pyrazole-4-carbaldehyde le (2 g, 20.814 mmol) and K2CO3 (7.19 g, 52.035 mmol) in ACN (10 mL) were added CD3I (4.16 g, 31.221 mmol) dropwise at room temperature. The resulting mixture was heated to 60°C and continued for 2h. The mixtureAttorney Reference: ALLN-001WOwas cooled to room temperature, filtered, the filter cake was washed with EtOAc (3 x 100 mL). The combined filtrates were concentrated under reduced pressure. The residue was purified by reversed -phase flash column chromatography using the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. Removal of solvents afforded l-(methyl-d3)-lH-pyrazole-4-carbaldehyde If (2 g, 84.93%) as a light yellow oil. LC-MS (ESI): m / z [M+H]+= 114.09.Step 5:

[0120] To a stirred solution of l-(methyl-d3)-lH-pyrazole-4-carbaldehyde If (2 g, 17.678 mmol) and propanoic acid (1.57 g, 21.214 mmol) in pyridine (10 mL) was added piperidine (4.52 g, 53.034 mmol) dropwise at room temperature under a nitrogen atmosphere. The resulting mixture was heated to and stirred at 110°C for 6h. After cooling to rt, the mixture was concentrated under reduced pressure to a residue, added water (10 mL), and acidified to pH 3 with conc. HCl. The product 1g was precipitated. The precipitated solids were collected by filtration and washed with water (3 x 50 mL) to give (£’)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)acrylic acid 1g (2.1 g, 76.55%) as a light yellow solid. LC-MS (ESI): m / z [M+H]+= 156.10. Step 6:

[0121] To a stirred solution of (E)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)acrylic acid 1g (100 mg, 0.644 mmol) and tert-butyl (R)-((l-(piperidin-3-ylmethyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate Id (247.47 mg, 0.837 mmol) in DMF (2 mL) were added DIEA (416.47 mg, 3.220 mmol), EDCI (150.07 mg, 0.966 mmol) and HOBT (130.62 mg, 0.966 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2h. The mixture was purified by reversed-phase flash column chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. Removal of solvents provided tert-butyl (R, E')-((1-((1-(3-(l-(methyl-d3)-lH-pyrazol-4-yl)acryloyl)piperidin-3-yl)methyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate Ih (220 mg, 78.92%) as a white solid. LC-MS (ESI): m / z [M+H]+= 432.27.Step 7:

[0122] A solution of tert-butyl (R,E)-((1-((1-(3-(1-(methyl-d3)-1H-pyrazol-4-yl)acryloyl)piperidin-3-yl)methyl)-1H-1,2,3-triazol-4-yl)methyl)carbamate 1h (300 mg, 0.694 mmol) in a 2.0 M solution of HCl in 1,4-dioxane (6 mL) was stirred at room temperature for Ih. The resulting mixture was concentrated under reduced pressure, then diluted with water (5 mL), basified to pH 7 with a saturated Na2CC>3 aq. solution. The residue was purified by reversed-phase flash chromatography with the following conditions: column, Cl 8 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UVAttorney Reference: ALLN-001WO254 nm. After lyophilization, the titled product (R,E)-1-(3-((4-(aminomethyl)-1H-1,2,3-triazol-1-yl)methyl)piperidin-1-yl)-3-(1-(methyl-d3)-1H-pyrazol-4-yl)prop-2-en-1-one Ex.1 was obtained (60.1 mg, 26.07%) as a white solid.1H NMR (400 MHz, DMSO-d6) 5 (ppm): 7.81 - 8.00 (m, 3H), 7.35 (d, J= 15.2 Hz, 1H), 6.77 - 6.94 (m, 1H), 4.25 - 4.39 (m, 2H), 4.15 - 4.2 (m, 1H), 3.84 - 3.99 (m, 1H), 3.84 (s, 2H), 3.06 - 3.18 (m, 1H), 2.68 - 2.94 (m, 1H), 1.87 - 2.12 (m, 1H), 1.52 - 1.83 (m, 2H), 1.14 - 1.45 (m, 2H LC-MS (ESI): m / z [M+H]+= 333.15.Example 2: Preparation of (R,E)-1-(3-((4-(aminomethyl)-1H-1,2,3-triazol-1-yl)methyl-d2)piperidin-1-yl)-3-(1-methyl-1H-pyrazol-4-yl)prop-2-en-1-oneStep 1:

[0123] To a stirred solution of (37?)-l-(teit-butoxycarbonyl)piperidine-3-carboxylic acid 2a (3 g, 13.085 mmol) and Et3N (1.99 g, 19.628 mmol) in THF (30 mL) at -20°C under nitrogen atmosphere was added 2-methylpropyl chloroformate (2.68 g, 19.628 mmol, 1.5). The resulting mixture was then stirred at room temperature for Ih. To the above mixture was added methanol-d4 (21 mL) followed by sodium tetrahydroborate-d4 (0.84 g, 20.020 mmol, 1.53) in portions over 10 min at 0°C. The reaction mixture was stirred at room temperature for an additional 2.5h. The reaction progress was monitored by LCMS. The mixture was adjusted to pH 8 using a saturated NaHCO3 (aq.) solution. The aqueous mixture was extracted with EtOAc (3 x 50mL). The combined organic layers were washed with brine (3x50 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to a residue, which was purified by silica gel column chromatography, eluted with CH2Cl2 / MeOH (10:1) to afford tert-butyl (R)-3-(hydroxymethyl-d2)piperidine-1-carboxylate 2b (2.5 g, 87.92%) as a yellow oil. LC-MS (ESI): m / z [M+H]+= 218.00.Step 2:Attorney Reference: ALLN-001WO

[0124] To a stirred solution of tert-butyl (R)-3-(hydroxymethyl-d2)piperidine-1-carboxylate 2b (2.5 g, 11.505 mmol) in 1,4-dioxane (10 mL) was added HC1 in 1,4-dioxane (4.0 M, 10 mL) at room temperature. The resulting mixture was stirred at room temperature for Ih, and the progress was monitored by LCMS. The mixture was concentrated under reduced pressure to give (R)-piperidin-3-ylmethan-d2-ol 2c (1.9 g) as a white solid. The crude product mixture was used in the next step directly without further purification. LC-MS (ESI): m / z [M+H]+= 118.00. Step 3:

[0125] To a stirred solution of (2E)-3-(l-methylpyrazol-4-yl)prop-2-enoic acid 2d (500 mg, 3.286 mmol) and (R)-piperidin-3-ylmethan-d2-ol 2c (577.65 mg, 4.929 mmol ) in DMF (5 mL) at room temperature under a nitrogen atmosphere was added EDCI (765.24 mg, 4.929 mmol), DIEA (2123.64 mg, 16.430 mmol) and HOBT (666.07 mg, 4.929 mmol). The resulting mixture was stirred at room temperature for 2h. The reaction progress was monitored by LCMS. The mixture was filtered, the filter cake was washed with EtOAc (3 x 100 mL). The filtrate was concentrated under reduced pressure to a residue, which was purified by reversed-phase flash column chromatography using the following conditions: column, Cl 8 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. Solvents were removed under vacuo to give (E)-1-(3-(hydroxymethyl-d2)piperidin-1-yl)-3-(1-methyl-1H-pyrazol-4-yl)prop-2-en-1-one 2e (340 mg, 41.17%) as a white solid. LC-MS (ESI): m / z [M+H]+= 252.00.Step 4:

[0126] To a stirred solution of (E, E)-l-(3-(hydroxymethyl-d2)piperidin-l-yl)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en- 1 -one 2e (340 mg, 1.353 mmol) and DPPA (446.76 mg, 1.624 mmol) in toluene (3.5 mL) was added DBU (247.15 mg, 1.624 mmol) at room temperature under a nitrogen atmosphere. The resulting mixture was heated at 110°C for 2h. The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature and concentrated under reduced pressure. The residue was neutralized to pH 7 with a saturated Na2CO3 (aq.) solution. The aqueous mixture was extracted with EtOAc (3 x lOOmL). The combined organic extracts were washed with brine (3 x 100 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2Cl2 / MeOH (10:1) to afford (E)-1-(3-(azidomethyl-d2)piperidin-1-yl)-3-(1-methyl-1H-pyrazol-4-yl)prop-2-en-1-one 2f (300 mg, 80.25%) as a yellow green oil. LC-MS (ESI): m / z [M+H]+= 277.00.Step 5:

[0127] To a stirred solution of (E, E)-l-(3-(azidomethyl-d2)piperidin-l-yl)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en- 1 -one 2f (300 mg, 1.086 mmol) and tert-butyl N-(prop-2-yn-l-Attorney Reference: ALLN-001WOyl)carbamate (219.03 mg, 1.412 mmol) in t-BuOH (2.5 mL) and H2O (1.5 mL) was added sodium (2S)-2-[(lS)-l,2-dihydroxyethyl]-4-hydroxy-5-oxooxolan-3-olate (43.01 mg, 0.217 mmol) and CuSO4.5H2O (54.21 mg, 0.217 mmol) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2h. The mixture was filtered, the filter cake was washed with EtOAc (3 x 50 mL), and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2CI2 / MeOH (10:1) to afford tert-butyl (R,E)-((1-((1-(3-(1-methyl-1H-pyrazol-4-yl)acryloyl)piperidin-3-yl)methyl-d2)-1H-1,2,3-triazol-4-yl)methyl)carbamate 2g (210 mg, 44.83%) as a colorless oil. LC-MS (ESI): m / z [M+H]+= 432.00.Step 6:

[0128] To a stirred solution of tert-butyl (R,E)-((1-((1-(3-(1-methyl-1H-pyrazol-4-yl)acryloyl)piperidin-3-yl)methyl-d2)-1H-1,2,3-triazol-4-yl)methyl)carbamate 2g (150 mg, 0.348 mmol) in Dioxane (2 mL) was added HCl in 1,4-dioxane (4.0 M, 2 mL) at 0°C. The resulting mixture was stirred at room temperature for 2h, while the progress was monitored by LCMS. The mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash column chromatography using the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% HC1), 10% to 50% gradient in 10 min; detector, UV 254 nm. After lyophilization the titled product ( / ?,£’)-l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl-d2)piperidin-l-yl)-3-(l -methyl- lH-pyrazol-4-yl)prop-2-en-l -one Ex.2 was obtained (50.9 mg, 44.18%) as a white solid. 4 INMR (300 MHz, DMSO-d6) 5 (ppm):8.19 (s, 1H), 8.03 (s, 1H), 7.84 (s, 1H), 7.32 - 7.37 (d, 1H), 6.89 - 6.94 (d, 1H), 4.06 - 4.13 (m, 4H), 3.83 - 3.86 (s, 3H), 2.90 - 3.13 (m, 2H), 1.98 - 2.06 (m, 1H), 1.69 - 1.70 (m, 2H), 1.28 - 1.31 (m, 2H); LC-MS (ESI): m / z [M+H]+= 332.15.Attorney Reference: ALLN-001WOExample 3: Preparation of (R,E)-1-(3-((4-(aminomethyl)-1H-1,2,3-triazol-1-yl)methyl)piperidin-1-yl-2,2,6,6-d4)-3-(1-methyl-1H-pyrazol-4-yl)prop-2-en-1-oneStep 1:

[0129] To a stirred solution of ( / ?)-piperidin-3-yl methanol 3a (10 g, 77.424 mmol) and HOAc (6.97 g, 116.136 mmol) in H2O (30 mL) was added NaNCL (8.01 g, 116.095 mmol) at 0°C. The resulting mixture was stirred at room temperature overnight. The reaction was quenched with Na2COs at 0°C. The mixture was stirred at room temperature for another 5 h, and extracted with CH2Cl2(3 x 100 mL). The combined organic layers were washed with brine (2 x 200 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to afford (R)-(1-nitrosopiperidin-3-yl)methanol 3b (12 g) as a brown solid. This crude product was used in the next step directly without further purification. LC-MS (ESI): m / z [M+H]+= 145.10.Step 2:

[0130] A solution of (R)-(1-nitrosopiperidin-3-yl)methanol 3b (10 g, 63.228 mmol) and NaOD (aq.) (25 mL) in D2O (50 mL) was stirred at 100°C overnight. The reaction mixture was allowed to cool down to room temperature, and extracted with CH2Cl2(3 x 100 mL). The combined organic layers were washed with brine (2 x 200 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to give a crude product of (R)-(1-nitrosopiperidin-3-yl-2,2,6,6-d4)methanol 3c (5 g) as a yellow oil, used without purification in the next step. The crude product was used in the next step directly without further purification. LC-MS (ESI): m / z [M+H]+= 149.20.Attorney Reference: ALLN-001WOStep 3:

[0131] A mixture of (R)-(l-nitrosopiperidin-3-yl-2,2,6,6-d4)methanol 3c (5 g, 37.542 mmol) and [2- Aluminium-Nickel Alloy (5 g) in Deuterium oxide (20 mL) was stirred at 80°C for 4h. The mixture was allowed to cool down to room temperature, and filtered. The filter cake was washed with D2O (2 x 50 mL). The combined filtrate was acidified to pH 6 using cone. HC1, then concentrated under reduced pressure. The residue was dissolved in MeOH (20 mL), filtered, the filter cake was washed with MeOH (2 x 10 mL). The combined methanol filtrates were concentrated under reduced pressure to give (R)-(piperidin-3-yl-2,2,6,6-d4)methanol 3d (3.5 g, 78.21%) as a yellow solid. LC-MS (ESI): m / z [M+H]+= 120.15.Step 4:

[0132] A mixture of aldehyde 3e (10.48 g, 95.1 mmol) and piperidine (243 mg, 2.85 mmol) was taken up in 50 mL of pyridine followed by the addition of malonic acid (9.9 g, 95 mmol) in one portion. The mixture was stirred at 110° C under N2 for 6 hours, and then room temperature for overnight. The solution was diluted with 125 mL of water, cooled in an ice bath, and then treated dropwise with 55 mL of concentrated HC1 to a pH = 1. The precipitate was collected by filtration and the filter cake was washed with water, hexane, and then dried under high vacuum at 45° C for 1 hour to afford the acrylic acid 3f (5.5 g, 38%).1HNMR (DMSO-de) 5 (ppm): 12.04 (s, 1H), 8.05 (s, 1H), 7.81 (s, 1H), 7.43 (d, 7=16 Hz, 1H), 6.16 (d, 7=16 Hz, 1H), 3.81 (s, 3H).Step 5:

[0133] To a stirred solution of (E)-3-(l-methyl-lH-pyrazol-4-yl)acrylic acid 3f (350 mg, 2.300 mmol) and (R)-(piperidin-3-yl-2,2,6,6-d4)methanol 3d (329.04 mg, 2.760 mmol) in DMF (4 mL) were added HATU (1311.99 mg, 3.450 mmol) and DIEA (1486.54 mg, 11.500 mmol) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2 h, and quenched with H2O (10 mL). The aqueous mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with brine (3 x 30 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by reversed-phase flash column chromatography with the following conditions: column, Cl 8 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. Removal of solvents afforded (R, E)-l-(3-(hydroxymethyl)piperidin-l-yl-2,2,6,6-d4)-3-(l -methyl- lH-pyrazol-4-yl)prop-2-en-l -one 3g (418 mg, 71.73%) as a yellow semi-solid. LC-MS (ESI): m / z [M+H]+= 254.15.Step 6:

[0134] To a stirred solution of (R, E)-l-(3-(hydroxymethyl)piperidin-l-yl-2,2,6,6-d4)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one 3g (300 mg, 1.184 mmol) in toluene (10 mL) wereAttorney Reference: ALLN-001WOadded DBU (252.39 mg, 1.658 mmol) and DPPA (456.25 mg, 1.658 mmol, 1.4 equiv) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 110°C for 2 h. After cooling to rt, the mixture was concentrated under vacuum. The residue was dissolved in H2O (10 mL), extracted with EA (3 x 10 mL). The combined organic layers were washed with water (2 x 30 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to a residue which was purified by silica gel column chromatography, eluted with CI bCb / MeOI I (12:1) to afford ( / ?, E)-l-(3-(azidomethyl)piperidin-l-yl-2,2,6,6-d4)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one 3h (135 mg, 40.96%) as a yellow semi-solid. LC-MS (ESI): m / z [M+H]+= 279.15.Step 7:

[0135] To a solution of (7?,£)-l-(3-(azidomethyl)piperidin-l-yl-2,2,6,6-d4)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en- 1 -one 3h (135 mg, 0.485 mmol ) in t-BuOH (2.5 mL) and H2O (1.5 mL) was treated with tert-butyl N-(prop-2-yn-l-yl)carbamate (97.85 mg, 0.630 mmol) at room temperature, followed by the addition of CuSO4.5H2O (24.22 mg, 0.097 mmol) at room temperature. To the above mixture was added Sodium ascorbate (19.22 mg, 0.097 mmol). The resulting mixture was stirred at room temperature for 2 h. H2O (10 mL) was added, the aqueous mixture was extracted with EA (3 x 10 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CfhCh / MeOH (9:1) to provide tert-butyl ( / ?,£’)-((l-((l-(3-(l-methyl-lH-pyrazol-4-yl)acryloyl)piperidin-3-yl-2,2,6,6-d4)methyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate 3i (150 mg, 71.34%) as a white solid.Step 8:

[0136] To a solution of tert-butyl ( / ?,£)-((l-((l-(3-(l-methyl-lH-pyrazol-4-yl)acryloyl)piperidin-3-yl-2,2,6,6-d4)methyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate 3i (150 mg, 0.346 mmol) in 1,4-dioxane (3 mL) was treated with HC1 in 1,4-dioxane (2.0 M, 3 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h, then concentrated under vacuum. The residue was basified to pH 8 using a saturated Nal ICO? (aq.), and further purified by reversed-phase flash column chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. After lyophilization, the titled product (R, E)-1-(3-((4-(aminomethyl)- 1H-1,2,3-triazol- 1 -yl)methyl)piperidin- l-yl-2, 2,6, 6-d4)-3-(l -methyl- 1H-pyrazol-4-yl)prop-2-en-l-one Ex.3 (69.1 mg, 59.90%) was obtained as a white solid. ‘HNMR (300 MHz, CD3OD) 8 (ppm): 7.88 (s, 2H), 7.80 (s, 1H), 7.44 (d, J = 15.0 Hz, 1H), 6.61 - 7.03 (m, 1H), 4.23 - 4.55 (m, 2H), 3.75 - 4.01 (m, 5H), 1.99 - 2.37 (m, 1H), 1.64 - 1.99 (m, 2H), 1.10 - 1.64 (m, 2H LC-MS (ESI): m / z [M+H]+= 334.20.Attorney Reference: ALLN-001WOExample 4: Preparation of (E, E)-l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one-2,3-d2Step 1:

[0137] To a stirred solution of 4-ethynyl-l -methyl- IH-pyrazole 4a (2 g, 18.845 mmol) in THF (15 mL) was added n-BuLi (1.45 g, 22.614 mmol) dropwise at -78°C under a nitrogen atmosphere. The reaction mixture was stirred at -78 °C for 30min. To the above mixture was added methyl chloroformate (3.56 g, 37.690 mmol) dropwise over 10 min at -78°C. The reaction was continued at -78°C for an additional Ih, and quenched with water at 0°C. The aqueous mixture was extracted with EtOAc (3 x 100 mL). The organic extracts were concentrated under reduced pressure to a residue which was purified by silica gel column chromatography, eluted with CI FCF / McOI I (12:1) to provide methyl 3-(l-methyl-lH-pyrazol-4-yl)propiolate 4b (2.5 g, 80.81%) as a yellow solid. LC-MS (ESI): m / z [M+H]+= 165.05.Step 2:

[0138] To a stirred solution of methyl 3-(l-methyl-lH-pyrazol-4-yl)propiolate 4b (2.5 g, 15.229 mmol) in Deuterium oxide (10 mL) and THF (100 mL) was added PPh3 (7.99 g, 30.458 mmol) in portions at room temperature. The resulting mixture was heated at 85°C for 2h. The mixture was allowed to cool down to room temperature, and extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (2 x 100 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by reversed-phase flash column chromatography with the following conditions: column, Cl 8 silica gel; mobile phase, McCN in Water (0.1% NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm to afford methyl (E)-3-(l -methyl- lH-pyrazol-4-yl)acrylate-d24c (600 mg, 23.43%) as a yellow solid, and also methyl (Z)-3-(l-methyl-lH-pyrazoL4-yl)acrylate-d24d (500 mg, 19.52%) as a yellow solid. LC-MS (ESI): m / z [M+H]+= 169.10. The geometric configuration of 4c (E) andAttorney Reference: ALLN-001WO4d (Z) was determined by comparing spectroscopic data of 4c and 4d to their corresponding protio analogs that were generated using the same method.Step 3:

[0139] A mixture of methyl (E)-3-(l-mcthyl-lH-pyrazol-4-yl)acrylatc-d24c (600 mg, 3.567 mmol) and LiOH (170.88 mg, 7.134 mmol) in H2O (2 mL), THF (2 mL), and methanol (2 mL) was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure. The residue was dissolved in water (10 mL), acidified to pH 3 with HC1 (aq.). The resulting aqueous mixture was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (2 x 50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to afford (E)-3-(l-methyl-lH-pyrazol-4-yl)acrylic-2,3-d2 acid 4e (500 mg, 90.92%) as a yellow solid. LC-MS (ESI): m / z [M+H]+= 155.07.Step 4:

[0140] To a solution of (£)-3-(l-methyl-lH-pyrazol-4-yl)acrylic-2,3-d2 acid 4e (100 mg, 0.649 mmol) in DMF (2 mL) was added tert-butyl ( / ?)-(( l-(piperidin-3-ylmethyl)-l H-1,2,3-triazol-4-yl)methyl)carbamate Id (249.09 mg, 0.844 mmol) at room temperature for 1 min followed by the addition of DIEA (419.18 mg, 3.245 mmol). HATU (369.96 mg, 0.974 mmol) was added. The resulting mixture was stirred at room temperature for 2 h. The reaction progress was monitored by LCMS. The mixture was filtered, the filter cake was washed with EA (3 x 20 mL). The filtrates were concentrated under reduced pressure to a residue which was purified by reversed-phase flash column chromatography with the following conditions: column, Cl 8 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 0% to 25% gradient in 10 min; detector, UV 254 nm. Removal of solvents afforded tert-butyl (R, E)-((l-((l-(3-(l -methyl- 1H-pyrazol-4-yl)acryloyl-d2)piperidin-3-yl)methyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate 4f (200 mg, 71.45%) as a white solid. LC-MS (ESI): m / z [M+H]+= 432.25Step 5:

[0141] To a stirred solution of tert-butyl (R, E)-((l-((l-(3-(l-methyl-lH-pyrazol-4-yl)acryloyl-d2)piperidin-3-yl)methyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate 4f (200 mg, 0.463 mmol) in 1,4-dioxane (3.0 mL) was added HC1 in 1,4-dioxane (2.0 M, 3 mL) dropwise at room temperature. The resulting mixture was sti rred at room temperature for 2h and concentrated under reduced pressure. The residue was basified to pH 8 with saturated Nal ICO (aq.) solution, and purified by reversed-phase flash column chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. After lyophilization the titled product ( / ?, / ?)-! -(3-((4-( aminomethyl)- 1 H- 1,2,3-triazoL 1 -yl)methyl)piperidin-l -yl)-3-( 1 -methyl- 1 H-pyrazol-4-yl)prop-2-en-l-one-2,3-d2 Ex.4 was obtained (105.3 mg, 68.55%) as a white solid. 'H-NMR (300 MHz,Attorney Reference: ALLN-001WOCD3OD) 5 (ppm): 7.93 (s, 1H), 7.88 (s, 1H), 7.79 (s, 1H), 4.25 - 4.52 (m, 2H), 4.10 - 4.27 (m, 1H), 3.91 - 4.10 (m, 2H), 3.58 - 3.91 (m, 4H), 2.63 - 3.22 (m, 2H), 1.95 - 2.40 (m, 1H), 1.61 -1.95 (m, 2H), 1.01 - 1.61 (m, 2H); LC-MS (ESI): m / z [M+H]+= 332.20.Example 5: Preparation of (R, Z)-l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one-2,3-d2Step 1:

[0142] To a stirred solution of methyl (Z)-3-(l-methyl-lH-pyrazol-4-yl)acrylate-d24d (500 mg, 2.976 mmol), available from Preparative Example 4, in H2O (4 mL), THF (4 mL), and methanol (4 mL) was added LiOH (142.8 mg, 5.952 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h, and then concentrated under reduced pressure. The residue was dissolved in water (20 mL), acidified to pH 3 with HC1 (aq.). The aqueous mixture was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (2 x 50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure, resulted in (Z)-3-(l-methyl-lH-pyrazoL4-yl)acrylic-2,3-d2 acid 5a (368 mg, 89.28%) as a yellow solid. LC-MS (ESI): m / z [M+H]+= 155.07.Step 2:

[0143] To a solution of (Z)-3-(l-methyl-lH-pyrazol-4-yl)acrylic-2,3-d2 acid 5a (150 mg, 0.973 mmol) in DMF (2 mL) was added tert-butyl ( / ?)-((l-(piperidin-3-ylmethyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate Id (373.40 mg, 1.265 mmol) at room temperature for 1 min followed by the addition of DIEA (627.58 mg, 4.865 mmol). HATU (554.61 mg, 1.460 mmol) was added. The resulting mixture was stirred at room temperature for an additional 2 h. The mixture was filtered, the filter cake was washed with EA (3 x 20 mL). The combined filtrates were concentrated under reduced pressure. The residue was purified by reversed-phase flash column chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 0% to 25% gradient in 10 min; detector, UV 254 nm.Attorney Reference: ALLN-001WORemoval of solvents provided tert-butyl (R, Z)-((l-((l-(3-(l-methyl-lH-pyrazol-4-yl)acryloyl-d2)piperidin-3-yl)methyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate 5b (350 mg, 83.33%) as a white solid. LC-MS (ESI): m / z [M+H]+= 432.25.Step 3:

[0144] To a stirred solution of tert-butyl (R, Z)-((l-((l-(3-(l-methyl-lH-pyrazol-4-yl)acryloyl-d2)piperidin-3-yl)methyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate 5b (350 mg, 0.812 mmol, 1 equiv) in 1,4-dioxane (3.0 mL) was added HC1 in 1,4-dioxane (2.0 M, 3 mL) dropwise at room temperature. The resulting mixture was stirred for 2h. The reaction progress was monitored by LCMS. The mixture was concentrated under reduced pressure, the residue was basified to pH 8 with saturated Nal [('() (aq.) and 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 50% gradient in 10 min; detector, UV 254 nm. After removal of solvents the titled product (R, Z)-l-(3-((4-(aminomethyl)-lII-l,2,3-triazol-l-yl)methyl)piperidin-l-yl)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one-2,3-d2 Ex.5 was obtained (65.8 mg, 24.48%) as a white solid. 'II-NMR (300 MHz, CD3OD) 5 (ppm): 7.90 (s, 1H), 7.76 (s, 1H), 7.53 (d, J =15.0 Hz, 1H), 4.19 - 4.46 (m, 3H), 3.76 - 4.00 (m, 6H), 2.75 - 3.23 (m, 2H), 1.85 - 2.28 (m, 1H), 1.60 - 1.85 (m, 2H), 1.22 - 1.59 (m, 2H LC-MS (ESI): m / z [M+H]+= 332.15.Example 6: (R, E)-l-(3-((4-(aminomethyl-d2)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl)- 3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-oneStep 1:

[0145] To a stirred solution of benzyl (3R)-3-(azidomethyl)piperidine-l -carboxylate lb (1.38 g, 5.031 mmol) and methyl propiolate (0.55 g, 6.540 mmol) in H2O (6 mL) and t-BuOH (10 mL) was added Sodium ascorbate (0.20 g, 1.006 mmol) and CuSO4.5H2O (0.25 g, 1.006 mmol) atAttorney Reference: ALLN-001WOroom temperature. The resulting mixture was stirred for 2h. Reaction progress was monitored by LCMS. The reaction was quenched by the addition of water (20 mL). The aqueous mixture was extracted with EtOAc (3 x 50mL). The combined organic layers were washed with brine (3 x 50 mL), dried over anhydrous ISteSCU, and concentrated under reduced pressure to a residue, which was purified by silica gel column chromatography, eluted with DCM / MeOH (5: 1) to afford benzyl (3R)-3-{ [4-(methoxycarbonyl)-l, 2, 3-triazol-l-yl]methyl}piperi dine- 1 -carboxylate 6a (1.2 g, 66.56 %) as a white solid. LC-MS (ESI): m / z [M+H]+= 359.00.Step 2:

[0146] To a stirred solution of benzyl (3R)-3-{ [4-(methoxycarbonyl)-l,2,3-triazol-l-yl]methyl [piperidine- 1 -carboxylate 6a (1.2 g, 3.348 mmol) in CD3OD (12 mL) was added NaBD4 (0.28 g, 6.696 mmol) at room temperature. The resulting mixture was heated and stirred at 70°C overnight. Reaction progress was monitored by LCMS. The mixture was allowed to cool down to room temperature, then quenched with ice water. The aqueous mixture was filtered, the filter cake was washed with EtOAc (3 x 20 mL). The filtrate was concentrated under reduced pressure to a residue, which was purified by silica gel column chromatography, eluted with CILCh / MeOH (5:1) to provide benzyl (3R)-3-({4-[hydroxy(2H2)methyl]-l,2,3-triazol-l-yl}methyl)piperidine-l -carboxylate 6b (860 mg, 77.27 %) as a white solid. LC-MS (ESI): m / z [M+H]+= 333.20.Step 3:

[0147] To a stirred solution of benzyl (3R)-3-({4-[hydroxy(2H2)methyl]-l,2,3-triazol-l-yl}methyl)piperidine-l -carboxylate 6b (840 mg, 2.527 mmol) and DPPA (834.55 mg, 3.032 mmol) in toluene (8 mL) was added DBU (461.67 mg, 3.032 mmol) dropwise at room temperature under a nitrogen atmosphere. The resulting mixture was heated to and stirred at 110°C for 2h. The reaction progress was monitored by LCMS. The mixture was allowed to cool down to room temperature, and quenched by the addition of water (20 mL). The aqueous mixture was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (3x20 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with C bCh / MeOH (5:1) to afford benzyl (3R)-3-({4-[azido(2H2)methyl]-l,2,3-triazol-l-yl}methyl)piperidine-l-carboxylate 6c (750 mg, 83.04%) as a yellow oil. LC-MS (ESI): m / z [M+H]+= 358.22.Step 4:

[0148] To a stirred solution of benzyl (3R)-3-({4-[azido(2H2)methyl]-I,2,3-triazol-l-yl}methyl)piperidine-l -carboxylate 6c (750 mg, 2.098 mmol) in HFIP (7 mL) was added A1CL (839.34 mg, 6.294 mmol) at room temperature under a nitrogen atmosphere. The resulting mixture was slirred for 2h. The reaction progress was monitored by LCMS. The reaction wasAttorney Reference: ALLN-001WOquenched with ice water. The aqueous mixture was filtered, the filter cake was washed with MeOH (3 x 20 mL). The filtrate was concentrated under reduced pressure to a residue, which was purified by reversed-phase flash column chromatography with the following conditions: column, C18 silica gel; mobile phase, McCN in Water (10 mmol / T NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. Removal of solvents afforded (3R)-3-({4-razido(2H2)methyl]-l,2,3-triazol-l-yl}methyl)piperidine 6d (360 mg, 76.84%) as a brown oil. TC-MS (ESI): m / z [M+H]+= 224.00.Step 5:

[0149] To a stirred solution of (2E)-3-(l-methylpyrazol-4-yl)prop-2-enoic acid 3f (200 mg, 1.314 mmol) and (3R)-3-({4-[azido(2H2)methyl]-l,2,3-triazol-l-yl}methyl)piperidine 6d (350.24 mg, 1.569 mmol) in DMF (3 mL) was added DIEA (849.45 mg, 6.570 mmol) and HATU (749.71 mg, 1.971 mmol) at room temperature. The resulting mixture was stirred for 2h. The reaction progress was monitored by LCMS. The reaction mixture was filtered, the filter cake was washed with EtOAc (3 x 20 mL). The combined filtrates were concentrated under reduced pressure to a residue, which was purified by reversed-phase flash column chromatography with the following conditions: column, Cl 8 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. Removal of solvents gave (2E)-l-[(3R)-3-({4-[azido(2H2)methyl]-l,2,3-triazol-l-yl}methyl)piperidin-l-yl]-3-(l-methylpyrazol-4-yl)prop-2-en-l-one 6e (90 mg, 19.16%) as a colorless oil. LC-MS (EST): m / z [M+H]+= 358.15.Step 6:

[0150] To a stirred solution of (2E)-l-[(3R)-3-({4-[azido(2H2)methyl]-l,2,3-triazol-l-yl}methyl)piperidin-l-yl]-3-(l-methylpyrazol-4-yl)prop-2-en-l-one 6e (90 mg, 0.252 mmol) and PPh? (79.26 mg, 0.302 mmol) in THF (0.8 mL) and H2O (0.2 mL) was added KOH (35.32 mg, 0.630 mmol) at room temperature. The resulting mixture was stirred for Ih. Water (50 mL) was added, and the aqueous mixture was extracted with EtOAc (3 x 50mL). The combined organic layers were washed with brine (3 x 150 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by reversed-phase flash column chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. Removal of solvents afforded the titled product (2£’)-l-[(3R)-3-({4-[amino(2H2)rnethyl]-l,2,3-triazol-l-yl}methyl)piperidin-l-yl]-3-(l-methylpyrazol-4-yl)prop-2-en-l-one Ex.6 (39.0 mg, 46.73%) as a white solid. 'HNMR (400 MHz, CD3OD) 5 (ppm): 7.94 (s, IH), 7.88 (s,lH), 7.80 (s,lH), 7.44 (d, J= 15.2 Hz, IH), 6.76 - 6.92 (m, IH), 4.20 - 4.40 (m, 2H), 4.15 - 4.20 (m, IH), 3.95 - 4.15Attorney Reference: ALLN-001WO(m, 1H), 3.80 - 3.90 (m, 3H), 3.20 - 3.30 (m, 1H), 2.80 - 3.10 (m, 1 H), 2.10 - 2.30 (m, 1 H), 1.71 - 1.88 (m, 2 H), 1.30 - 1.65 (m, 2 H); LC-MS (ESI): m / z [M+H]+= 332.20.Example 7: (R, E)-l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl-d2)piperidin-l-yl)- 3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-oneStep 1:

[0151] To a stirred solution of (E)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)acrylic acid 1g (500 mg, 3.222 mmol) in DMF (5 mL) was added (R)-piperidin-3-ylmethan-d2-ol 2c (755.22 mg, 6.444 mmol) and D1EA (1.84 g, 14.236 mmol) at room temperature under a nitrogen atmosphere. HATU (1837.82 mg, 4.833 mmol) was added. The resulting mixture was stirred for 2h.Reaction progress was monitored by LCMS. The mixture was filtered, the filter cake was washed with EtOAc (3 x 50 mL). The combined filtrates were concentrated under reduced pressure. The residue was purified by reversed-phase flash column chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. Removal of solvents afforded (R,£)-l-(3-(hydroxymethyl-d2)piperidin-l-yl)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one 7a (250 mg, 30.50%) as a yellow semi-solid. LC-MS (ESI): m / z [M+H]+= 255.20.Step 2:

[0152] To a stirred solution of (R, E)-l-(3-(hydroxymethyl-d2)piperidin-l-yl)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one 7a (230 mg, 0.904 mmol) in toluene (5 mL) was added DBU (165.20 mg, 1.085 mmol) and DPPA (298.63 mg, 1.085 mmol) dropwise at room temperature under a nitrogen atmosphere. The resulting mixture was heated to and stirred at 110°C for an additional 2h. The mixture was concentrated under reduced pressure. The residue was dissolved in water (10 mL). The aqueous mixture was extracted with EtOAc (3 x 10 mL).Attorney Reference: ALLN-001WOThe combined organic extracts were dried over anhydrous Na2SC>4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with DCM / MeOH (10:1) to give (R, E)-l-(3-(azidomethyl-d2)piperidin-l-yl)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-cn- 1 -one 7b (180 mg, 71.25%) as a yellow viscous oil. LC-MS (ESI): m / z [M+H]+= 280.18.Step 3:

[0153] To a stirred solution of ( / ?, E)-l-(3-(azidomethyl-d2)piperidin-l-yl)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one 7b (160 mg, 0.573 mmol) and tert-butyl N-(prop-2-yn-l-yl)carbamate (115.55 mg, 0.745 mmol) in water (1.5 mL) and t-BuOH (2.5 mL) was added sodium ascorbate (113.47 mg, 0.573 mmol) and CuSC)4.5H2O (28.60 mg, 0.115 mmol) in portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred for 2h. The reaction progress was monitored by LCMS. The mixture was filtered, the filter cake was washed with EtOAc (3 x 100 mL). The filtrate was concentrated under reduced pressure to a residue, which was purified by silica gel column chromatography, eluted with DCM / MeOH (10:1) to give tert-butyl (7?, E)-((l-((l-(3-(l-(methyl-d3)-lH-pyrazol-4-yl)acryloyl)piperidin-3-yl)methyl-d2)-lH-l,2,3-triazol-4-yl)methyl)carbamate 7c (130 mg, 52.23%) as a light yellow solid. LC-MS (ESI): m / z [M+H]+= 435.25.Step 4:

[0154] To a stirred solution of tert-butyl (E, E)-((l-((l-(3-(l-(methyl-d3)-lH-pyrazol-4-yl)acryloyl)piperidin-3-yl)methyl-d2)-lH-l,2,3-triazol-4-yl)methyl)carbamate 7c (110 mg, 0.253 mmol) was added HC1 in 1,4-dioxane (2.0 M, 6 mL) dropwise at room temperature. The resulting mixture was stirred for 2h, then concentrated under reduced pressure. The residue was neutralized to pH 7 with saturated Na2CO3 (aq.), and 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 50% gradient in 10 min; detector, UV 254 nm. After lyophilization the titled product (7?, E)-l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl-d2)piperidin-l-yl)-3-(l-(methyl-d )-lH-pyrazol-4-yl)prop-2-en-l-one Ex.7 (56.6 mg, 66.86%) was obtained as a white semi-solid.1HNMR (300 MHz, CD3OD) 5 (ppm): 7.95 (s, 1H), 7.89 (s, 1H), 7.80 (s, 1H), 7.45 (d, J = 15.0 Hz, 1H), 6.76 - 6.93 (m, 1H), 4.15 - 4.29 (m, 1H), 3.89 - 4.11 (m, 3H). 3.06 - 3.16 (m. 1H), 2.74 - 2.98 (m, 1H), 2.01 - 2.25 (m, 1H). 1.71 - 1.91 (m. 2H), 1.46 - 1.81 (m, 2H); LC-MS (ESI): m / z [M+H]+= 335.15.Attorney Reference: ALLN-001WOExample 8: (R,E)-1-(3-((4-(aminomethyl)-1H-1,2,3-triazol-1-yl)methyl)piperidin-1-yl-2,2,6,6-d4)-3-(1-(methyl-d3)-1H-pyrazol-4-yl)prop-2-en-1-oneStep 1:

[0155] To a solution of (E)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)acrylic acid 1g (300 mg, 1.933 mmol) in DMF (3 mL) was added (3R)-(2,2,6,6-2H4)piperidin-3-ylmethanol 3d (345.68 mg, 2.90 mmol) at room temperature under a nitrogen atmosphere, followed by the addition of DIEA (1249.40 mg, 9.665 mmol). To the above mixture was added HATU (1102.69 mg, 2.900 mmol). The resulting mixture was stirred for 2 h. The reaction progress was monitored by LCMS. The reaction was quenched with water (50 mL). The aqueous mixture was extracted with EA (3 x 50 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by reversed-phase flash column chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 0% to 25% gradient in 10 min; detector, UV 254 nm. Removal of solvents provided (R,£)-l-(3-(hydroxymethyl)piperidin-l-yl-2,2,6,6-d4)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one 8a (312 mg, 61.03) as a yellow oil. LC-MS (ESI): m / z [M+H]+= 257.15.Step 2:

[0156] To a solution of (R, E)-l-(3-(hydroxymethyl)piperidin-l-yl-2,2,6,6-d4)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one 8a (290 mg, 1.131 mmol) in toluene (8 mL) was added DPPA (435.85 mg, 1.583 mmol) at room temperature under a nitrogen atmosphere, followed by the addition of DBU (241.11 mg, 1.583 mmol). The resulting mixture was heated to and stirred at 110°C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure. The residue was dissolved in water (50 mL), and extracted with EA (3 x 100 mL). The combined organic layers were washed with brine (2 x 100 mL), dried over anhydrous Na2SO4,Attorney Reference: ALLN-001WOand concentrated under reduced pressure to a residue, which was purified by silica gel column chromatography, eluted with CH2Cl2 / MeOH (9:1) to afford ( / ?, E)-l-(3-(azidomethyl)piperidin-l-yl-2,2,6,6-d4)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one 8b (100 mg, 31.42%) as a yellow semi-solid. LC-MS (ESI): m / z [M+H]+= 282.20.Step 3:

[0157] To a solution of (R, E)-l-(3-(azidomethyl)piperidin-l-yl-2,2,6,6-d4)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one 8b (100 mg, 0.355 mmol) in t-BuOH (2 mL) and H2O (1.5 mL) was added tert-butyl N-(prop-2-yn-l-yl)carbamate (71.70 mg, 0.461 mmol) at room temperature, followed by the addition of CuSO4.5H2O (17.75 mg, 0.071 mmol). To the above mixture was added sodium ascorbate (14.08 mg, 0.071 mmol). The resulting mixture was stirred at room temperature for 2h. H2O (10 mL) was added. The aqueous mixture was extracted with EA (3 x 10 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2Cl2 / MeOH (9:1) to provide tert-butyl (R, E)-((l-((l-(3-(l-(methyl-d3)-lH-pyrazol-4-yl)acryloyl)piperidin-3-yl-2,2,6,6-d4)methyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate 8c (89 mg, 57.36%) as a white solid. LC-MS (ESI): m / z [M+H]+= 437.25.Step 4:

[0158] To a stirred solution of tert-butyl (R,£’)-((l-((l-(3-(l-(methyl-d3)-lH-pyrazol-4-yl)acryloyl)piperidin-3-yl-2,2,6,6-d4)methyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate 8c (85 mg, 0.195 mmol) in 1,4-dioxane (2 mL) was added HCl in 1,4-dioxane (2.0M, 2 mL) at room temperature. The resulting mixture was stirred for 2 h, then concentrated under reduced pressure to a residue. The residue was basified to pH 8 with saturated NaHCO3 (aq.), and 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 50% gradient in 10 min; detector, UV 254 nm. Removal of solvents afforded the titled product (R, E)-l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl-2,2,6,6-d4)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one Ex.8 (32.8 mg, 50.07%) as a colorless solid. 'HNMR (300 MHz, CD3OD) 6 (ppm): 7.88 (s, 2H), 7.80 (s, 1H), 7.44 (d, J = 15.0 Hz, 1H), 6.68 - 7.05 (m, 1H), 4.20 - 4.57 (m, 2H), 3.90 (s, 2H), 2.00 - 2.32 (m, 1H), 1.63 - 2.00 (m, 2H), 1.19 - 1.63 (m, 2H LC-MS (ESI): m / z [M+H]+= 337.25.Attorney Reference: ALLN-001WOExample 9: (7?, E)-l-(3-((4-(aminomethyl-d2)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl)- 3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-oneStep 1:

[0159] To a solution of (£’)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)acrylic acid 1g (900 mg, 5.800 mmol) in DMF (10 mL) was added (3R)-piperidin-3-ylmethanol (1002.04 mg, 8.700 mmol) at room temperature followed by the addition of DIEA (2248.92 mg, 17.400 mmol). To the above mixture was added HATU (3308.07 mg, 8.700 mmol). The resulting mixture was stirred for an additional 2 h. Reaction progress was monitored by LCMS. The mixture was filtered: the filter cake was washed with EtOAc (3 x 100 mL). The filtrate was concentrated under reduced pressure to a residue, which was purified by reversed-phase flash column chromatography with the following conditions: column, Cl 8 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 0% to 25% gradient in 10 min; detector, UV 254 nm. Removal of solvents gave (R, E)-l-(3-(hydroxymethyl)piperidin-l-yl)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one 9a (920 mg, 62.86%) as a white solid. LC-MS (ESI): m / z [M+H]+= 253.20.Step 2:

[0160] To a solution of (R, E)-l-(3-(hydroxymethyl)piperidin-l-yl)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one 9a (430 mg, 1.704 mmol) in toluene (8 mL) was added DPPA (656.57 mg, 2.386 mmol) at room temperature followed by the addition of DBU (363.21 mg, 2.386 mmol). The resulting mixture was heated to and stirred at 110°C for 2 h. Reaction progress was monitored by LCMS. The reaction mixture was concentrated under reducedAttorney Reference: ALLN-001WOpressure. The residue was dissolved in water (50 mL), and the aqueous mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with brine (2 x 100 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with DCM / MeOH (9:1) to provide (R, E)-l-(3-(azidomethyl)piperidin-l-yl)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one 9b (466 mg, 98.60%) as a yellow oil. LC-MS (ESI): m / z [M+H]+= 278.20.Step 3:

[0161] To a solution of (R,E)-1-(3-(azidomethyl)piperidin-1-yl)-3-(1-(methyl-d3)-1H-pyrazol-4-yl)prop-2-en-1-one 9b (460 mg, 1.659 mmol) in t-BuOH (5 mL) and H2O (3 mL) was added methyl propiolate (181.28 mg, 2.157 mmol) at room temperature followed by the addition of CuSO4.5H2O (82.82 mg, 0.332 mmol). To the above mixture was added Sodium ascorbate (65.71 mg, 0.332 mmol). The resulting mixture was stirred for 2 h, and quenched by the addition of H2O (5mL). The aqueous mixture was extracted with EA (3 x 10 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with DCM / MeOH (9:1) to give methyl ( / ?, E)-l-((l-(3-(l-(methyl-d3)-lH-pyrazol-4-yl)acryloyl)piperidin-3-yl)methyl)-lH-l,2,3-triazole-4-carboxylate 9c (500 mg, 83.41%) as a white solid. LC-MS (ESI): m / z [M+H]+= 362.20.Step 4:

[0162] To a stirred solution of methyl ( / ?, E)-l-((l-(3-(l-(methyl-d3)-lH-pyrazol-4-yl)acryloyl)piperidin-3-yl)methyl)-lH-l,2,3-triazole-4-carboxylate 9c (200 mg, 0.553 mmol) in THF (3 mL) was added Lithium aluminum deuteride (25.55 mg, 0.608 mmol) dropwise at room temperature. The resulting mixture was stirred for 1 h. The reaction was quenched by the addition of Na2SO4.10H2O (100 mg) at room temperature. The resulting mixture was filtered, the filter cake was washed with EA (3 x 20 mL). The filtrate was concentrated under reduced pressure to afford (R,£)-l-(3-((4-(hydroxymethyl-d2)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one 9d (150 mg, 80.81%) as a white solid. LC-MS (ESI): m / z [M+H]+= 336.25.Step 5:

[0163] To a solution of (R,£)-l-(3-((4-(hydroxymethyl-d2)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one 9d (150 mg, 0.447 mmol) in toluene (5 mL) was added DPPA (172.30 mg, 0.626 mmol) at room temperature followed by the addition of DBU (95.31 mg, 0.626 mmol). The resulting mixture was stirred at 110°C for 2 h. Reaction progress was monitored by LCMS. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in water (50 mL). The aqueous mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed withAttorney Reference: ALLN-001WObrine (2 x 100 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with DCM / MeOH (9:1) to provide (R,E)-1-(3-((4-(azidomethyl-d2)-1H-1,2,3-triazol-1-yl)methyl)piperidin-1-yl)-3-(1-(methyl-d3)-1H-pyrazol-4-yl)prop-2-en-1-one 9e (100 mg, 62.04%) as a dark yellow oil. LC-MS (ESI): m / z [M+H]+= 361.25.Step 6:

[0164] To a stirred solution of (R, E)-l-(3-((4-(azidomethyl-d2)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one 9e (100 mg, 0.277 mmol) in THF (2 mL) and H2O (0.5 mL) was added KOH (38.91 mg, 0.693 mmol) followed by PPh3 (87.32 mg, 0.332 mmol) in portions at room temperature. The resulting mixture was stirred for 1 h, and then 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% TEA), 10% to 50% gradient in 10 min; detector, UV 254 nm. Removal of solvents afforded the titled product (R, E)-l-(3-((4-(aminomethyl-d2)-lH-l,2,3-triazol- 1 -yl)methyl)piperidin- 1 -y 1) -3 -( 1 -(methyl-d3)- 1 H-pyrazol-4-yl)prop-2-en- 1 -one Ex.9 (22.3 mg, 24.03%) as a light yellow solid. 'HNMR (300 MHz, CD3OD) 5 (ppm): 7.88 (s, 2H), 7.79 (s, 1H), 7.44 (d, J =15.3 Hz, 1H), 6.51 - 7.03 (m, 1H), 4.29 - 4.60 (m, 2H), 3.82 - 4.29 (m, 2H), 2.66 - 3.21 (m, 2H), 2.01 - 2.29 (m, 1H). 1.65 - 2.00 (m, 2H), 1.30 - 1.65 (m, 2H); LC-MS (ESI): m / z [M+H]+= 335.20.Attorney Reference: ALLN-001WOExample 10: (Zf, E) -(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)inetIiyl)piperi(iin-l-yl)-3- (l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one-2,3-d2Step 1:

[0165] To a stirred solution of 4-ethynyl-lH-pyrazole 10a (2 g, 21.715 mmol) and K2CO3 (7.50 g, 54.288 mmol) in MeCN (20 mL) was added CD3I (4.72 g, 32.572 mmol) dropwise at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 60 °C for 2h. The mixture was cooled to room temperature, filtered, and the filter cake was washed with MeCN (3 x 30 mL). The combined filtrates were concentrated under reduced pressure to give a crude product, 4-ethynyl-l-(methyl-d3)-lH-pyrazole 10b (1.5 g, 63.29%) as a white solid. 10b was used in the next step directly without purification. LC-MS (ESI): m / z [M+H]+= 110.05. Step 2:

[0166] To a stirred solution of 4-ethynyl-l-(methyl-d3)-lH-pyrazole 10b (1.6 g, 14.659 mmol) in THF (15 mL) at -78°C under a nitrogen atmosphere was added n-BuLi (1.13 g, 17.591 mmol) dropwise. The resulting mixture was stirred at -78 °C for 30 min. To the above mixture was added methyl chloroformate (2.77 g, 29.318 mmol) dropwise over 10 min at -78°C. The reaction mixture was stirred at -78 °C for an additional 2h. The reaction was quenched with the addition of water at 0°C. The aqueous mixture was extracted with EtOAc (3 x 100 mL). The organicAttorney Reference: ALLN-001WOextracts were concentrated under reduced pressure to a residue, which was purified by silica gel column chromatography, eluted with CH2Cl2 / MeOH (12:1) to afford methyl 3-(1-(methyl-d3)-1H-pyrazol-4-yl)propiolate 10c (1.2 g, 48.96%) as a yellow solid. LC-MS (ESI): m / z [M+H]+= 168.05.Step 3:

[0167] To a stirred solution of methyl 3-(l-(methyl-d3)-lH-pyrazol-4-yl)propiolate 10c (1.2 g, 7.178 mmol) in Deuterium oxide (10 mL) and THF (100 mL) were added PPh3 (7.99 g, 30.458 mmol) at room temperature. The resulting mixture was heated to and stirred at 85 °C for 2h. The mixture was cooled down to room temperature, and extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (2 x 100 mL), dried over anhydrous Na2SO4, and 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% NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm to provide two geometric isomers, methyl (E)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)acrylate-d2 lOd (490 mg, 39.87%) as a yellow solid, and methyl (Z)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)acrylate-d2 lOe (400 mg, 32.55%) as a yellow solid. The geometric configuration of the trans- and cis- alkenyl moiety was assigned based the comparison of spectra data of these two products to their protio analogs synthesized under the same conditions. lOd: LC-MS (ESI): m / z [M+H]+= 172.10; lOe: LC-MS (ESI): m / z [M+H]+= 172.10.Step 4:

[0168] To a stirred solution of methyl (E)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)acrylate-d2 lOd (200 mg, 1.169 mmol) in a mixture of solvents H2O (1 mL), THF (1 mL) and MeOH (1 mL) was added LiOH (56 mg, 2.339 mmol) at room temperature. The resulting mixture was stirred for 2 h and concentrated under reduced pressure. The residue was dissolved in water (10 mL), acidified to pH 3 with HC1 (aq.). The resulting aqueous mixture was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (2 x 50 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to give (E)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)acrylic-2,3-d2 acid lOf (160 mg, 87.43%) as a yellow solid. LC-MS (ESI): m / z [M+H]+= 158.05.Step5:

[0169] To a solution of (E)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)acrylic-2,3-d2 acid lOf (100 mg, 0.637 mmol) in DMF (2 mL) was added tert-butyl ( / ?)-(( l-(piperidin-3-ylmethyl)-lH- 1,2,3-triazol-4-yl)methyl)carbamate Id (250 mg, 0.828 mmol) followed by the addition of DIEA (419.18 mg, 3.245 mmol) at room temperature. To the above mixture was added HATU (370 mg, 0.955 mmol). The reaction mixture was stirred for additional 2 h. The mixture was filtered,Attorney Reference: ALLN-001WOthe filter cake was washed with EA (3 x 20 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 (10 mmol / L NH4HCO3), 0% to 25% gradient in 10 min; detector, UV 254 nm. Removal of solvents generated tert-butyl (R, E)-((l-((l-(3-(l-(methyl-d3)-lH-pyrazol-4-yl)acryloyl-d2)piperidin-3-yl)methyl)-lH-l,2,3- triazol-4-yl)methyl)carbamate 10g (200 mg, 71.45%) as a white solid. LC-MS (ESI): m / z [M+H]+= 435.25.Step 6:

[0170] To a stirred solution of tert-butyl (R, E)-((l-((l-(3-(l-(methyl-d3)-lH-pyrazol-4- yl)acryloyl-d2)piperidin-3-yl)methyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate 10g (200 mg, 0.460 mmol) in 1,4-dioxane (4 mL) was added HC1 in 1,4-dioxane (2.0 M, 4 mL) dropwise at room temperature. The resulting mixture was stirred for 2h. The reaction progress was monitored by LCMS. The mixture was concentrated under reduced pressure, basified to pH 8 with saturated NaHCO3 (aq.). and 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 50% gradient in 10 min; detector, UV 254 nm. After lyophilization, the titled product (R,£)-l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl)-3-(l- (methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one-2,3-d2 Ex.10 (93.5 mg, 60.74%) was obtained as a white solid. ¹HNMR (300 MHz, CD3OD) δ (ppm): 7.93 (s, 1H). 7.88 (s, 1H), 7.79 (s, 1H), 3.70 - 4.66 (m, 6H), 2.65 - 3.26 (m, 2H), 2.00 - 2.50 (m, 1H), 1.65 - 2.00 (m, 2H), 1.21 - 1.65 (m, 2H); LC-MS (ESI): m / z [M+H]+= 335.25.Example 11: (R,Z)-1-(3-((4-(aminomethyl)-1H-1,2,3-triazol-1-yl)methyl)piperidin-1-yl)-3-(1-(methyl-d3)-1H-pyrazol-4-yl)prop-2-en-1-one-2,3-d2Step 1:Attorney Reference: ALLN-001WO

[0171] To a stirred solution of methyl (Z)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)acrylate-d2 lOe (200 mg, 1.169 mmol), available from Preparative Example 10, in a mixture of solvents H2O (1 mL), THF (1 mL) and methanol (1 mL) was added LiOH (56 mg, 2.339 mmol) at room temperature. The resulting mixture was stirred for 2 h and concentrated under reduced pressure. The residue was dissolved in water (10 mL), acidified to pH 3 with HC1 (aq.), and extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (2 x 50 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to provide (Z)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)acrylic-2,3-d2acid Ila (150 mg, 81.52%) as a yellow solid. LC-MS (ESI): m / z [M+H]+= 158.05.Step 2:

[0172] To a solution of (Z)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)acrylic-2,3-d2 acid Ila (100 mg, 0.637 mmol) in DMF (2 mL) was added tert-butyl (R)-((l-(piperidin-3-ylmethyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate Id (250 mg, 0.828 mmol) at room temperature followed by the addition of DIEA (419.18 mg, 3.245 mmol). To the above mixture was added HATU (370 mg, 0.955 mmol). The resulting mixture was stirred for an additional 2 h. The reaction progress was monitored by LCMS. The mixture was filtered, the filter cake was washed with EA (3 x 20 mL). The filtrate was concentrated under reduced pressure to a residue, which was purified by reversed- phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 0% to 25% gradient in 10 min; detector, UV 254 nm. Removal of solvents provided tert-butyl (7?, Z)-((l-((l-(3-(l-(methyl-d3)-lH-pyrazol-4-yl)acryloyl-d2)piperidin-3-yl)methyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate lib (180 mg, 84.90%) as a white solid. LC-MS (ESI): m / z [M+H]+= 435.25.Step 3:

[0173] To a stirred solution of tert-butyl ( / ?, Z)-((l-((l-(3-(l-(methyl-d3)-lH-pyrazol-4-yl)acryloyl-d2)piperidin-3-yl)methyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate 11b (180 mg, 0.460 mmol) in 1,4-dioxane (4 mL) were added HC1 in 1,4-di oxane (2.0 M, 4 mL) dropwise at room temperature. The resulting mixture was stirred for 2h, then concentrated under reduced pressure. The mixture was basified to pH 8 with saturated NaHCO3 (aq.), and 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 50% gradient in 10 min; detector, UV 254 nm. After lyophilization, the titled product (7?, Z)-l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl)-3-(l-(methyl-d3)-lH-pyrazol-4-yl)prop-2-en-l-one-2,3-d2Ex.ll (113.4 mg, 81.88%) was obtained as a white solid. ¹HNMR (300 MHz, CD3OD) δ (ppm): 7.94 (s, 1H), 7.78 (s, 1H), 7.55 (s, 1H), 4.31 - 4.45 (m, 2H), 4.20 - 4.30 (m, 1H), 3.90 -Attorney Reference: ALLN-001WO4.02 (m, 2H), 3.75 - 3.89 (m, 1H). 2.71 - 3.25 (m, 2H), 1.62 - 2.21 (m, 3H), 1.21 - 1.61 (m, 2H): LC-MS (ESI): m / z [M+H]+= 335.20.Example 12: (R, E)-l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl- 2,2,6,6-d4)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one-2,3-d2Step 1:

[0174] To a stirred solution of (E)-3-( 1 -methyl- lH-pyrazol-4-yl)acrylic-2,3-d2 acid 4e (100 mg, 0.649 mmol) available from Preparative Example 4, and (R)-(piperidin-3-yl-2,2,6,6-d4)methanol 3d (154.64 mg, 1.298 mmol) available from Preparative Example 3, in DMF (2 mL) was added DIEA (419.18 mg, 3.245 mmol) and HATU (369.96 mg, 0.974 mmol) at room temperature. The resulting mixture was stirred for 2 h. The reaction progress was monitored by LCMS. The mixture was filtered, the filter cake was washed with ethyl acetate (3 x 20 mL), and the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase flash column chromatography with the following conditions: column, Cl 8 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. Removal of solvents gave (R,£)-l-(3-(hydroxymethyl)piperidin-l-yl-2,2,6,6-d4)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one-2,3-d212a (120 mg, 72.45%) as a yellow oil. LC-MS (ESI): m / z [M+H]+= 256.15.Step 2:

[0175] To a solution of (R,£)-l-(3-(hydroxymethyl)piperidin-l-yl-2,2,6,6-d4)-3-(l-methyl- lH-pyrazol-4-yl)prop-2-en-l -one-2, 3-d212a (120 mg, 0.470 mmol) in toluene (5 mL) was added DPPA (181.06 mg, 0.658 mmol) at room temperature under a N2 atmosphere, followed byAttorney Reference: ALLN-001WOthe addition of DBU (100.16 mg, 0.658 mmol). The resulting mixture was heated to and stirred at 110°C for 2 h. The reaction was monitored by LCMS. The mixture was concentrated under reduced pressure. The residue was dissolved in water (50 mL). The aqueous mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with brine (2 x 100 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with DCM / MeOH (9:1) to afford ( / / <)-!-(3-(azidomethyl)piperidin-l-yl-2,2,6,6-d4)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one-2,3-d2 12b (100 mg, 75.90%) as a white solid. LC-MS (ESI): m / z [M+H]+= 281.20.Step3:

[0176] To a solution of (R,£’)-l-(3-(azidomethyl)piperidin-l-yl-2,2,6,6-d4)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l -one-2, 3-d212b (100 mg, 0.357 mmol) in t-BuOH (2.5 mL) and H2O (1.5 mL) was added tert-butyl N-(prop-2-yn-l-yl)carbamate (71.96 mg, 0.464 mmol) at room temperature, followed by the addition of CuSO4.5H2O (17.81 mg, 0.071 mmol). To the above mixture was added Sodium ascorbate (14.13 mg, 0.071 mmol). The resulting mixture was stirred for an additional 3 h. H2O (5 mL) was added; the aqueous mixture was extracted with EA (3 x 10 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with DCM / MeOH (9: 1) to give tertbutyl ( / ?, E')-((l-((l-(3-(l-niethyl-lH-pyrazol-4-yl)acryloyl-d2)piperidin-3-yl-2,2,6,6-d4)niethyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate 12c (100 mg, 64.37%) as a white solid. LC-MS (ESI): m / z [M+H]+= 436.25.Step 4:

[0177] To a stirred solution of tert-butyl (R, E)-((l-((l-(3-(l-methyl-lH-pyrazol-4-yl)acryloyl-d2)piperidin-3-yl-2,2,6,6-d4)methyl)-lH-l,2,3-triazol-4-yl)methyl)carbamate 12c (100 mg, 0.230 mmol) in dioxane (2 mL) was added HC1 in 1,4-dioxane (4.0 M, 2 mL) at room temperature. The resulting mixture was stirred for 2 h and concentrated under vacuum. The residue was basified to pH 8 with saturated Na2CO3 (aq.), and 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 50% gradient in 10 min; detector, UV 254 nm. Removal of solvents afforded the titled product (R, E)-l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl-2,2,6,6-d4)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one-2,3-d2 Ex.12 (36.7 mg, 47.65%) as a white solid.1HNMR (300 MHz, CD3OD) 5 (ppm): 7.88 (s, 2H), 7.79 (s, 1H), 4.16 - 4.60 (m, 2H), 3.71 -4.09 (m, 5H), 1.98 - 2.41 (m, 1H), 1.62 - 1.98 (m, 2H), 1.16 - 1.62 (m, 2H); LC-MS (ESI): m / z [M+H]+= 336.25.Attorney Reference: ALLN-001WOExample 13: (R, E')-l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl-d2)piperidin-l-yl)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one-2,3-d2Step 1:

[0178] To a stirred solution of (E)-3-(l-methyl-lH-pyrazol-4-yl)acrylic-2,3-d2 acid 4e (100 mg, 0.649 mmol) available from Preparative Example 4 and (R)-piperidin-3-ylmethan-d2-ol 2c (154.64 mg, 1.298 mmol) available from Preparative Example 2 in DMF (2 mL) was added DIEA (419.18 mg, 3.245 mmol) and HATU (369.96 mg, 0.974 mmol) at room temperature. The resulting mixture was stirred for 2 h. Reaction progress was monitored by LCMS. The mixture was filtered, the filter cake was washed with ethyl acetate (3 x 20 mL), and 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 50% gradient in 10 min; detector, UV 254 nm. Removal of solvents gave (R, E)-l-(3-(hydroxymethyl-d2)piperi din- l-yl)-3-(l -methyl- lH-pyrazol-4-yl)prop-2-en-l -one-2,3-d213a (120 mg, 72.45%) as a yellow oil. LC-MS (ESI): m / z [M+H]+= 254.15.Step 2:

[0179] To a solution of (R, E)-l-(3-(hydroxymethyl-d2)piperidin-l-yl)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l -one-2, 3-d213a (120 mg, 0.470 mmol) in toluene (5 mL) was added DPPA (181.06 mg, 0.658 mmol) at room temperature, followed by the addition of DBU (100.16 mg, 0.658 mmol) at room temperature under a N2 atmosphere. The resulting mixture was heated and stirred at 100°C for 2 h. The mixture was concentrated under the reduced pressure. The residue was dissolved in water (10 mL). The aqueous mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with brine (2 x 20 mL), dried over anhydrousAttorney Reference: ALLN-001WONa2SO4, an the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with DCM / MeOH (9:1) to provide (R, E)-l-(3-(azidomethyl-d2)piperidin-l-yl)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l -one-2, 3-d213b (100 mg, 75.90%) as a white solid. LC-MS (ESI): m / z [M+H]+= 279.20.Step 3:

[0180] To a solution of ( / ?, E’)-l-(3-(azidomethyl-d2)piperidin-l-yl)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one-2,3-d213b (100 mg, 0.357 mmol) in t-BuOH (2.5 mL) and H2O (1.5 mL) was added with tert-butyl N-(prop-2-yn-l-yl)carbamate (71.96 mg, 0.464 mmol) at room temperature, followed by the addition of CuSO4.5H2O (17.81 mg, 0.071 mmol). To the above mixture was added Sodium ascorbate (14.13 mg, 0.071 mmol). The reaction mixture was stirred for an additional 4 h. H2O (10 mL) was added; the aqueous mixture was extracted with EA (3 x 10 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with DCM / MeOH (9:1) to provide tert-butyl (R,E)-((1-((1-(3-(1-methyl-1H-pyrazol-4-yl)acryloyl-d2)piperidin-3-yl)methyl-d2)-1H-1,2,3-triazol-4-yl)methyl)carbamate 13c (110 mg, 66.55%) as a white solid. LC-MS (ESI): m / z [M+H]+= 434.25.Step 4:

[0181] To a stirred solution of tert-butyl ( / ?,£’)-((l-((l-(3-(l-methyl-lH-pyrazol-4-yl)acryloyl-d2)piperidin-3-yl)methyl-d2)-lH-l,2,3-triazol-4-yl)methyl)carbamate 13c (110 mg, 0.323 mmol) in 1,4-dioxane (2 mL) was treated with HC1 in 1,4-dioxane (2.0 M, 2 mL) at room temperature. The resulting mixture was stirred for 2 h and concentrated under vacuum. The residue was basified to pH 8 with saturated Na2CO? (aq.) and 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 50% gradient in 10 min; detector, UV 254 nm. After lyophilization, the titled product (R, E)-l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl-d2)piperidin-l-yl)-3-(l -methyl- lH-pyrazol-4-yl)prop-2-en-l -one-2, 3 -d2 Ex.13 (47.2 mg, 43.84%) was obtained as a white solid. 'HNMR (300 MHz, CD3OD) 5 (ppm): 7.88 (s, 2H), 7.79 (s, 1H), 4.00 - 4.37 (m, 2H), 3.72 - 4.00 (m, 5H), 2.73 - 3.28 (m, 2H), 2.00 - 2.36 (m, 1H), 1.66 - 2.00 (m, 2H), 1.15 - 1.66 (m, 2H); LC-MS (ESI): m / z [M+H]+= 334.15.Attorney Reference: ALLN-001WOExample 14: (R, E)- l-(3-((4-(aminomethyl-d2)-l H- l,2,3-triazol-l-yl)methyl)piperidin-l-yl)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one-2,3-d2Step 1:

[0182] To a stirred solution of (E)-3-(l-methyl-lH-pyrazol-4-yl)acrylic-2,3-d2 acid 4e (100 mg, 0.649 mmol) available from Preparative Example 4 and (R)-3-((4-(azidomethyl-d2)-1H-1,2,3-triazol-1-yl)methyl)piperidine 6d (217.25 mg, 0.974 mmol) available from Preparative Example 6 in DMF (3 mL) was added DIEA (419.18 mg, 3.245 mmol), followed by HATU (369.96 mg, 0.974 mmol) at room temperature. The resulting mixture was stirred for 2h. The reaction progress was monitored by LCMS. The mixture was filtered, the filter cake was washed with EtOAc (3 x 20 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 (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. Removal of solvents gave (R, E)-l-(3-((4-(azidomethyl-d2)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl)-3-(l -methyl- lH-pyrazol-4-yl)prop-2-en-l -one-2, 3-d214a (70 mg, 30.02%) as a white solid. LC-MS (ESI): m / z [M+H]+= 360.33.Step 2:

[0183] To a stirred solution of (R, E)-l-(3-((4-(azidomethyl-d2)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one-2,3-d214a (70 mg, 0.195 mmol) in THF (0.8 mL) and H2O (0.2 mL) was added PPh3 (61.30 mg, 0.234 mmol) and KOH (27.37 mg, 0.4875 mmol) at room temperature. The resulting mixture was stirred for Ih. The reaction progress was monitored by LCMS. The mixture was filtered, the filter cake was washed with EtOAc (3 x 20 mL). The combined filtrates were 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 50% gradient in 10 min; detector, UV 254 nm. Upon lyophilization, the titled product (R, E)-1 -(3-((4-(aminomethyl-d2)- 1 H- 1,2,3-triazoL 1 -yl)methyl)piperidin- 1 -y 1) -3 -( 1 -methyl- 1 H-Attorney Reference: ALLN-001WOpyrazol-4-yl)prop-2-en-l -one-2, 3-d2 Ex.14 (28.1 mg, 43.27%) was obtained as a white solid. ‘IINMR (400 MHz, CD30D) 5 (ppm): 7.88 (s, 2H), 7.80 (s,lH), 4.30 - 4.55 (m, 2H), 4.15 - 4.30 (m, 1H), 3.95 - 4.10 (m, 1H), 3.80 - 3.90 (m, 3H), 2.75 - 3.30 (m, 2H), 2.10 - 2.30 (m, 1H), 1.70 - 1.90 (m, 2H), 1.25 - 1.60 (m, 1H); LC-MS (ESI): m / z [M+H]+= 334.15.Example 15: (2?,£’)-l-(3-((4-(aminomethyl-d2)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl- 2,2,6,6-d4)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-oneStep 1:

[0184] To a stirred mixture of ( / ?, E)-l-(3-(azidomethyl)piperidin-l-yl-2,2,6,6-d4)-3-(l- methyl-lH-pyrazol-4-yl)prop-2-en-l-one 3h (420 mg, 1.509 mmol) available from Preparative Example 3 and propiolic acid methyl ester (164.91 mg, 1.962 mmol) in t-BuOH (4 mL) and H2O (2 mL) was added CuSO4.5H2O (75.35 mg, 0.302 mmol) and Sodium ascorbate (59.78 mg, 0.302 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The reaction progress was monitored by LCMS. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2Ch / MeOH (17:1) to afford methyl (7?, E)-l-((l-(3-(l-methyl-lH-pyrazol-4- yl)acryloyl)piperidin-3-yl-2,2,6,6-d4)methyl)-lH-l,2,3-triazole-4-carboxylate 15a (450 mg, 82.29%) as a colorless oil. LC-MS (ESI): m / z [M+H]+= 363.25.Step 2:

[0185] To a solution of methyl (7?, E)-l-((l-(3-(l-methyl-lH-pyrazol-4-yl)acryloyl)piperidin- 3-yl-2,2,6,6-d4)methyl)-lH-l,2,3-triazole-4-carboxylate 15a (450 mg, 1.242 mmol) in THF (5 mL) was treated with Lithium aluminum deuteride (62.55 mg, 1.490 mmol) at 0 °C for 5 minAttorney Reference: ALLN-001WOunder a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2 h. The reaction progress was monitored by LCMS. The reaction was quenched with the addition of Na2SO4.10H2O at room temperature. The mixture was filtered, the filter cake was washed with EtOAc (3 x 10 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase flash column chromatography with the following conditions: column, C18 silica gel: mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 0% to 30% gradient in 10 min; detector, UV 254 nm. Removal of solvents gave a crude product (400 mg), which was further purified by Prep-HPLC with the following conditions (Column: XSelect CSH Fluoro Pheny, 19*250 mm, 5pm; Mobile Phase A: Water(10mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 25 mL / min mL / min; Gradient (B%): 8% B to 30% B inlOmin; Wave Length: 254nm nm; RTl(min): 9.01) to afford (2E)-l-[(3S)-3-({4-[hydroxy(2H2)methyl]-l,2,3-triazol-l-yl}methyl)(2,2,6,6-2H4)piperidin-l-yl]-3-(l-methylpyrazol-4-yl)prop-2-en-l-one 15b (23.3 mg, 38.83%) as an off-white solid. LC-MS (ESI): m / z [M+H]+= 337.20.Step 3:

[0186] To a solution of(R, E)-l-(3-((4-(hydroxymethyl-d2)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl-2,2,6,6-d4)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one 15b (160 mg, 0.476 mmol) in toluene (2 mL) was treated with DBU (101.36 mg, 0.666 mmol) at room temperature followed by the addition of DPPA (183.24 mg, 0.666 mmol). The resulting mixture was heated and stirred at 110 °C for 2 h. The reaction progress was monitored by LCMS. The reaction mixture was concentrated under reduced pressure to a residue, which was purified by silica gel column chromatography, eluted with CH2Cl2 / MeOH (9:1) to afford (R, E)-l-(3-((4-(azidomethyl-d2)- 1 H- 1,2,3-triazol- 1 -yl)methyl)piperidin- 1 -yl-2,2,6,6-d4)-3-(l -methyl- 1H-pyrazol-4-yl)prop-2-en-l-one 15c (110 mg, 63.99%) as a dark grey oil. LC-MS (ESI): m / z [M+H]+= 362.30.Step 4:

[0187] To a solution of (R, E)-l-(3-((4-(azidomethyl-d2)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yl-2,2,6,6-d4)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one 15c (150 mg, 0.415 mmol) in THF (2 mL) and H2O (1.2 mL) was added KOH (58.21 mg, 1.037 mmol) followed by the addition of PPh3 (130.62 mg, 0.498 mmol) at room temperature. The resulting mixture was stirred for 2 h. The reaction progress was monitored by LCMS. The mixture was concentrated under reduced pressure to a residue, which was diluted with MeOH (3 mL) and purified by reversed-phase flash chromatography with the following conditions: column, Cl 8 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 0% to 20% gradient in 10 min; detector, UV 254 nm. After lyophilization, the titled product (R, E)-l-(3-((4-(aminomethyl-d2)-lH- 1,2, 3 -triazol- l-yl)methyl)piperidin-l -yl-2, 2,6, 6-d4)-3-(l -methyl- lH-pyrazol-4-yl)prop-Attorney Reference: ALLN-001WO2-en- 1-one Ex.15 (60.9 mg, 43.75%) was obtained as a white solid. 1HNMR (300 MHz, CD3OD) 5 (ppm): 7.89 (s, 1H), 8.06 (s, 1H), 7.85 (s, 1H), 7.80 (d, J = 15.3 Hz, 1H), 6.75 - 6.93 (m, 1H), 4.21 - 4.51 (m, 2H), 3.81 (s, 3H), 2.05 - 2.31 (m, 1H), 1.67 - 1.91 (m, 2H), 1.26 - 1.63 (m, 2H); LC-MS (ESI): m / z [M+H]+= 336.25.Example 16: (7f, E)-l-(3-((4-(aminomethyl-d2)-lH-l,2,3-triazol-l-yl)methyl-d2)piperidin-l- yl)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-oneStep 1:

[0188] To a solution of (R,£’)-l-(3-(azidomethyl-d2)piperidin-l-yl)-3-(l-methyl-lH-pyrazol-4- yl)prop-2-en-l-one 2f (700 mg, 2.533 mmol) available from Preparative Example 2 in t-BuOH (5 mL) and H2O (3 mL) was added methyl propiolate (276.86 mg, 3.293 mmol, 1.3) followed by the addition of CuSO4.5H2O (126.49 mg, 0.507 mmol) at room temperature. To the above mixture was added Sodium ascorbate (100.36 mg, 0.507 mmol). The resulting mixture was stirred for 2 h. The mixture was filtered, the filter cake was washed with EA (3 x 20 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 (10 mmol / L NH4HCO3), 0% to 10% gradient in 10 min; detector, UV 254 nm. Removal of solvents afforded methyl (R, E)-l-((l-(3-(l-methyl-lH-pyrazol-4- yl)acryloyl)piperidin-3-yl)methyl-d2)-lH-l,2,3-triazole-4-carboxylate 16a (620 mg, 67.91%) as a white solid. LC-MS (ESI): m / z [M+H]+= 361.20.Step 2:

[0189] To a stirred solution of methyl (R, E)-l-((l-(3-(l-methyl-lH-pyrazol-4- yl)acryloyl)piperidin-3-yl)methyl-d2)-lH-l,2,3-triazole-4-carboxylate 16a (600 mg, 1.665 mmol) in THF (6 mL) was added Lithium aluminum deuteride (76.87 mg, 1.832 mmol)Attorney Reference: ALLN-001WOdropwise at room temperature. The resulting mixture was stirred for 0.5 h. The reaction progress was monitored by TLC. The reaction was quenched by the addition of Na2SO4. IOH2O (100 mg) at room temperature. The mixture was filtered, the filter cake was washed with EA (3 x 20 mL). The filtrate was concentrated under reduced pressure to give (E, E)-l-(3-((4-(hydroxymethyl-d2)-lH-l,2,3-triazol-l-yl)methyl-d2)piperidin-l-yl)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one 16b (400 mg, 71.85%) as a light yellow solid. LC-MS (ESI): m / z [M+H]+= 335.25.Step 3:

[0190] To a solution of (E, E)-l-(3-((4-(hydroxymethyl-d2)-lH-l,2,3-triazol-l-yl)methyl-d2)piperi din- 1 -yl)-3-( 1 -methyl- lH-pyrazol-4-yl)prop-2-en-l -one 16b (400 mg, 1.196 mmol) in toluene (8 mL) was treated with DPPA (460.85 mg, 1.674 mmol) followed by the addition of DBU (254.94 mg, 1.674 mmol) at room temperature. The resulting mixture was heated and stirred at 110°C for 2 h. The reaction progress was monitored by LCMS. The mixture was concentrated under reduced pressure. The residue was dissolved in water (50 mL), and extracted with EA (3 x 100 mL). The combined organic layers were washed with brine (2 x 100 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2Cl2 / MeOH (9:1) to afford R, E)-l-(3-((4-(azidomethyl-d2)-lH-L2,3-triazol-l-yl)methyl-d2)piperidin-l-yl)-3-(l-methyl-lH-pyrazol-4-yl)prop-2-en-l-one 16c (300 mg, 69.78%) as a dark green semi-solid. LC-MS (ESI): m / z [M+H]+= 360.25.Step 4:

[0191] To a stirred solution of (E, E)-l-(3-((4-(azidomethyl-d2)-lH-l,2,3-triazol-l-yl)methyl-d2)piperi din- 1 -yl)-3-( 1 -methyl- lH-pyrazol-4-yl)prop-2-en-l -one 16c (200 mg, 0.556 mmol) in THF (5 mL) and H2O (3 mL) was added KOH (78.05 mg, 1.390 mmol) and PPh3 (175.14 mg, 0.667 mmol) in portions at room temperature. The resulting mixture was stirred at room temperature for 2h and 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 50% gradient in 10 min; detector, UV 254 nm. Upon lyophilization, the titled product (E, E)-l-(3-((4-(aminomethyl-d2)-lH-l,2,3-triazol-l-yl)methyl-d2)piperidin-l-yl)-3-(l -methyl- lH-pyrazoL4-yl)prop-2-en-l -one Ex.16 (43.1 mg, 23.23%) was obtained as a white solid. 1HNMR (300 MHz, CD3OD) 6 (ppm): 7.88 (s, 2H), 7.79 (s, 1H), 7.44 (d, J =15.3 Hz, 1H), 6.59 - 7.05 (m, 1H), 3.96 - 4.41 (m, 2H). 3.75 - 3.96 (m, 3H), 2.94 - 3.22 (m, 1H), 2.68 - 2.94 (m, 1H), 1.96 - 2.32 (m, 1H), 1.65 - 1.96 (m, 2H), 1.21 - 1.65 (m, 2H); LC-MS (ESI): m / z [M+H]+= 334.20.Attorney Reference: ALLN-001WOExample 17: ( / f, E')-l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl-d2)piperidin-l-yl)- 3-(l-(cyclopropylmethyl)-lH-pyrazol-4-yl)prop-2-en-l-oneStep 1:

[0192] To a solution of lH-pyrazole-4-carbaldehyde 17a (1 g, 10.407 mmol) in DMF (10 mL) was added (iodomethyl)cyclopropane (2.84 g, 15.611 mmol) at room temperature, followed by the addition of K2CO3 (3.59 g, 26.017 mmol). The resulting mixture was stirred at 60 °C for Ih, filtered, and concentrated. The residue was purified by reversed phase flash column chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 0% to 25% gradient in 30 min; detector, UV 254 nm. Removal of solvents provided 1 -(cyclopropylmethyl)- lH-pyrazole-4-carbaldehyde 17b (1.29 g, 82.54%) as a light yellow oil. LC-MS (ESI): m / z [M+H]+= 151.00.Step 2:

[0193] A solution of 1 -(cyclopropylmethyl)- lH-pyrazole-4-carbaldehy de 17b (1.24 g, 8.257 mmol) in pyridine (12 mL) was treated with malonic acid (1.03 g, 9.908 mmol) at room temperature under a nitrogen atmosphere, followed by the addition of piperidine (2.11 g, 24.771 mmol). The resulting mixture was heated to and stirred at 110 °C for 6 h under a nitrogen atmosphere. The reaction progress was monitored by LCMS. Upon cooling to room temperature, the reaction mixture was concentrated under reduced pressure. Water (100 mL) was added to the residue, the aqueous mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (2 x 200 mL), dried over anhydrous Na2SO4, and 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),Attorney Reference: ALLN-001WO0% to 25% gradient in 15 min; detector, UV 254 nm. Removal of solvents gave (E)-3-( 1 -(cyclopropylmethyl)- lH-pyrazol-4-yl)acry lie acid 17c (1 g, 63.01%) as a light yellow solid. LC-MS (ESI): m / z [M+H]+= 193.20.Step 3:

[0194] A solution of (2E)-3-[l-(cyclopropylmethyl)pyrazol-4-yl]prop-2-enoic acid 17c (500 mg, 2.601 mmol) in DMF (5 mL) was treated with (3R)-piperidin-3-yl(2H2)methanol 2c (396.28 mg, 3.381 mmol) at room temperature for 1 min followed by the addition of DIEA (1.68 g, 13.005 mmol). To the above mixture was added HATU (1.48 g, 3.901 mmol). The resulting mixture was stirred at room temperature for an additional 2 h. The mixture was filtered, the filter cake was washed with EA (3 x 20 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 (10 mmol / L NH4HCO3), 0% to 25% gradient in 10 min; detector, UV 254 nm. Removal of solvents afforded (2E)-3-[l-(cyclopropylmethyl)pyrazol-4-yl]-l-[(3R)-3-[hydroxy(2H2)methyl]piperidin-l-yl]prop-2-en-l-one 17d (330 mg, 43.54%) as a yellow viscous oil. LC-MS (ESI): m / z [M+H]+= 292.17.Step 4:

[0195] A solution of (R, E)-3-( 1 -(cyclopropylmethyl)- 1 H-pyrazol-4-yl)- 1 -(3- (hydroxy methyl-d2)piperidin-l-yl)prop-2-en-l-one 17d (330 mg, 1.132 mmol) in toluene (10 mL) was treated with DPPA (436.34 mg, 1.585 mmol) at room temperature for 1 min under a nitrogen atmosphere followed by the addition of DBU (241.38 mg, 1.585 mmol). The resulting mixture was heated to and stirred at 110°C for 2 h. Reaction progress was monitored by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in water (50 mL). The resulting aqueous mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (2 x 100 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2CI2 I MeOH (9:1) to provide (R, E)-l-(3-(azidomethyl-d2)piperidin-l-yl)-3-(l-(cyclopropylmethyl)-lH-pyrazol-4-yl)prop-2-en-l-one 17e (300 mg, 83.72%) as a yellow oil. LC-MS (ESI): m / z [M+H]+= 317.22.Step 5:

[0196] A solution of-(3-(azidomethyl-d2)piperidin- 1 -yl)-3-(l -(cyclopropylmethyl)-lH-pyrazol-4-yl)prop-2-en-l-one 17e (300 mg, 0.948 mmol) in t-BuOH (2 mL) was treated with tert-butyl N-(prop-2-yn-l-yl)carbamate (191.30 mg, 1.232 mmol) at room temperature followed by the addition of CuSO4.5H2O (47.35 mg, 0.190 mmol). To the above mixture was added sodium ascorbate (37.57 mg, 0.190 mmol). The resulting mixture was stirred at room temperature for 2 h, filtered, and the filter cake was washed with DCM (3x10 mL). TheAttorney Reference: ALLN-001WOcombined filtrates were concentrated under reduced pressure. The residue was purified by reversed-phase flash column chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 0% to 10% gradient in 10 min; detector, UV 254 nm to afford tert-butyl (R, E)-((l-((l-(3-(l-(cyclopropylmcthyl)-lH-pyrazol-4-yl)acryloyl)piperidin-3-yl)methyl-d2)-lH-l,2,3-triazol-4-yl)methyl)carbamate 17f (210 mg, 46.96%) as a white solid. LC-MS (ESI): m z [M+HJ+= 427.27.Step 6:

[0197] A solution of tert-butyl (R, E)-((1-((1-(3-(1-(cyclopropylmethyl)-1H-pyrazol-4-yl)acryloyl)piperidin-3-yl)methyl-d2)-1H-1,2,3-triazol-4-yl)methyl)carbamate 17f (200 mg, 0.424 mmol) in HC1 in 1,4-dioxane (2.0 M, 6 mL) was stirred at room temperature for 2h. The resulting mixture was concentrated under vacuum. The residue was basified to pH 8 with saturated Na2CC>3 (aq.), taken up and purified by reversed-phase flash column chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. Upon lyophilization, the titled product ( / ?, E)-l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl-d2)piperidin-l-yl)-3-(l-(cyclopropylmethyl)-lH-pyrazol-4-yl)prop-2-en-l-one Ex.17 (63.6 mg, 40.37%) was obtained as a white solid. ’I I-NMR (300 MHz, CD3OD) 5 (ppm): 8.00 (s, 1H), 7.80 (s, 2H), 7.46 (d, J =15.0 Hz, 1H), 6.65 - 7.10 (m, 1H), 3.73 - 4.46 (m, 6H), 2.69 - 3.27 (m, 2H), 2.00 - 2.38 (m, 1H), 1.69 - 2.00 (m, 2H), 1.15 - 1.69 (m, 3H), 0.50 - 0.80 (m, 2H), 0.25 -0.50 (m, 2H); LC-MS (ESI): m / z [M+H]+= 337.25.Example 18: (Z?, E')-l-(3-((4-(aminomethyl)-lH-l,2,3-triazol-l-yl)methyl)piperidin-l-yI- 2,2,6,6-d4)-3-(l-(cyclopropylmethyI)-lH-pyrazol-4-yI)prop-2-en-l-oneStep 1:

[0198] A solution of (2E)-3-[l-(cyclopropylmethyl)pyrazol-4-yl]prop-2-enoic acid 17c (300 mg, 1.561 mmol) available from Preparative Example 17, and (R)-(piperidin-3-yl-2,2,6,6-d₄)methanol 3d (279.06 mg, 2.341 mmol) in DMF (3 mL) was treated with DIEA (806.88 mg,Attorney Reference: ALLN-001WO6.244 mmol) at room temperature, followed by the addition of HATU (890.17 mg, 2.341 mmol). The resulting mixture was stirred for an additional 2 h. The reaction progress was monitored by LCMS. The mixture was concentrated and dissolved in water (50 mL). The aqueous mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (2 x 300 mL), dried over anhydrous Na2SO4, and 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), 0% to 25% gradient in 10 min; detector, UV 254 nm. Removal of solvents afforded (2£)-3-[l-(cyclopropylmethyl)pyrazol-4-yl]-l-[(3R)-3-(hydroxymethyl)(2,2,6,6-2d4)piperidin-l-yl]prop-2-en-l-one 18a (290 mg, 63.33%) as a yellow semi-solid. LC-MS (ESI): m / z [M+H]+= 294.25. Step 2:

[0199] A solution of (2E)-3-[1-(cyclopropylmethyl)pyrazol-4-yl]-1-[(3R)-3-(hydroxymethyl)(2,2,6,6-2d4)piperidin-1-yl]prop-2-en-1-one 18a (290 mg, 0.988 mmol) in toluene (3 mL) was treated with DPPA (326.41 mg, 1.186 mmol) at room temperature, followed by the addition of DBU (180.57 mg, 1.186 mmol). The resulting mixture was heated to and stirred at 110°C for 2 h. The reaction progress was monitored by LCMS. The mixture was concentrated under a reduced pressure. The residue was dissolved in water (50 mL). The aqueous mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (2 x 300 mL), dried over anhydrous Na2SO4, and concentrated under a reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2Cl2 / MeOH (9:1) to afford (2E)-1-[(3S)-3-(azidomethyl)(2,2,6,6-2d4)piperidin-1-yl]-3-[1-(cyclopropylmethyl)pyrazol-4-yl]prop-2-en-1-one 18b (140 mg, 44.48%) as a yellow oil. LCMS (ESI): m / z [M+H]+= 319.25.Step 3:

[0200] To a mixture of (2E)-1-[(3S)-3-(azidomethyl)(2,2,6,6-2d4)piperidin-1-yl]-3-[1-(cyclopropylmethyl)pyrazol-4-yl]prop-2-en-1-one 18b (140 mg, 0.471 mmol) and sodium ascorbate (18.66 mg, 0.094 mmol) in t-BuOH (2 mL) and H2O (1.2 mL) was added CuSO4.5H2O (23.52 mg, 0.094 mmol) at room temperature, followed by the addition of tertbutyl A-(prop-2-yn-l-yl)carbamate (95.04 mg, 0.612 mmol). The resulting mixture was stirred at room temperature for 2 h. Water (50 mL) was added. The aqueous mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (2 x 300 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, Cl 8 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 0% to 10% gradient in 10 min; detector, UV 254 nm. Removal of solvents provided tert-butyl N-[(1-{[(3S)-1-[(2E)-3-[1-Attorney Reference: ALLN-001WO(cyclopropylmethyl)pyrazol-4-yl]prop-2-enoyl](2,2,6,6-2H4)piperidin-3-yl]methyl}-l,2,3-triazol-4-yl)methyl]carbamate 18c (150 mg, 67.23%) as a yellow solid. LC-MS (ESI): m / z [M+H]+= 474.40.Step 4:

[0201] Into a 50 mL round-bottom flask was added tert-butyl N-[(1-{[(3S)-1-[(2E)-3-[1-(cyclopropylmethyl)pyrazol-4-yl]prop-2-enoyl](2,2,6,6-2H4)piperidin-3-yl]methyl}-1,2,3-triazol-4-yl)methyl]carbamate 18c (150 mg, 0.317 mmol) and HC1 in 1,4-dioxane (2.0 M, 4 mL) at room temperature. The resulting mixture was stirred at room temperature for 2h. The reaction progress was monitored by LCMS. The mixture was concentrated under reduced pressure, then diluted with water (5 mL). The aqueous residue was basified to pH 8 with saturated Na2CO3(aq.), and purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 0% to 25% gradient in 15 min; detector, UV 254 nm. Upon lyophilization, the titled product (2£)-l-[(3S)-3-{[4-(aminomethyl)-l,2,3-triazol-l-yl]methyl}(2,2,6,6-2H4)piperidin-l-yl]-3-[l-(cyclopropylmethyl)pyrazol-4-yl]prop-2-en-l-one Ex.18 (35.7 mg, 30.18%) as a white solid.1HNMR (300 MHz, CD3OD) δ (ppm): 8.00 (s, 1H), 7.92 (s, 1H), 7.82 (s, 1H), 7.47 (d, J= 15.3 Hz, 1H), 6.71 - 6.95 (m, 1H), 4.30 - 4.50 (m, 2H), 3.85 - 4.08 (m, 4H), 2.05 - 2.30 (m, 1H), 1.70 - 1.95 (m, 2H), 1.39 - 1.65 (m, 2H), 1.18 - 1.38 (m, 1H), 0.55 - 0.74 (m, 2H), 0.35 - 0.50 (m, 2H); LC-MS (ESI): m / z [M+H]+= 374.25.Attorney Reference: ALLN-001WOExample 19: (R, E)-1-(3-((4-(aminomethyl)-1H-1,2,3-triazol-1-yl)methyl)piperidin-1-yl-2,2,3,6,6-d5)-3-(1-methyl-1H-pyrazol-4-yl)prop-2-en-1-oneStep 1:

[0202] To a stirred solution of ( / ?)-piperidine-3 -carboxylic acid 19a (10 g, 77.519 mmol) and HOAc (7.01 g, 116.279 mmol) in H2O (30 mL) was added NaNO2(8.05 g, 116.279 mmol) at 0°C. The resulting mixture was stirred at room temperature overnight. The reaction was quenched with Na2COs at 0°C. The reaction mixture was stirred at room temperature for an additional 5 h. The aqueous mixture was extracted with CH2CI2 (3 x 100 mL). The combined organic layers were washed with brine (2 x 200 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to give (R)-1-nitrosopiperidine-3-carboxylic acid 19b (11 g) as a brown solid. This crude product was used in the next step directly without further purification. LC-MS (ESI): m / z [M+H]+= 159.10.Step 2:

[0203] A solution of (R)-1-nitrosopiperidine-3-carboxylic acid 19b (10 g, 63.228 mmol) and NaOD (aq.) (25 mL) in D2O (50 mL) was stirred at 100°C overnight. The mixture was allowed to cool down to room temperature and extracted with CH2Cl2(3 x 100 mL). The combined organic layers were washed with brine (2 x 200 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to provide (R)-1-nitrosopiperidine-3-carboxylic-2,2,3,6,6-d5 acid 19c (4.5 g) as a yellow oil. This crude product was used in the next step directly without further purification. LC-MS (ESI): m / z [M+H]+= 164.15.Step 3:Attorney Reference: ALLN-001WO

[0204] A mixture of (R)-l-nitrosopiperidine-3-carboxylic-2,2,3,6,6-d5 acid 19c (4.5 g, 33.582 mmol) and Aluminum-Nickel alloy in Deuterium oxide (20 mL) was stirred at 80°C for 4h. The mixture was allowed to cool down to room temperature and filtered, the filter cake was washed with D2O (2 x 50 mL). The filtrate was acidified to pH 5 with cone. HC1 and concentrated under reduced pressure. The residue was dissolved in MeOH (20 mL), filtered, the filter cake was washed with MeOH (2 x 10 mL). The combined filtrates were concentrated under reduced pressure to afford (R)-piperidine-3-carboxylic-2,2,3,6,6-ds acid 19d (3.3 g) as a yellow solid. LC-MS (ESI): m / z [M+H]+= 135.20.Step 4:

[0205] To a solution of (E)-3-(1-methyl-1H-pyrazol-4-yl)acrylic acid 2d (330 mg, 2.169 mmol) in DMF (5 mL) was added (R)-piperidine-3-carboxylic-2,2,3,6,6-d5 acid 19d (1164.16 mg, 8.676 mmol) at room temperature under nitrogen atmosphere, followed by the addition of DIEA (1401.60 mg, 10.845 mmol). To the above mixture was added HATU (1237.02 mg, 3.253 mmol). The resulting mixture was stirred for 2 h. The reaction progress was monitored by LCMS. H2O (10 mL) was added. The aqueous mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with brine (3 x 30 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, Cl 8 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 0% to 25% gradient in 10 min; detector, UV 254 nm. Removal of solvents afforded methyl (R,E)-1-(3-(1-methyl-1H-pyrazol-4-yl)acryloyl)piperidine-3-carboxylate-2,2,3,6,6-d5 19e (140 mg, 22.86%) as a colorless oil. LC-MS (ESI): m / z [M+H]+= 283.20.Step 5:

[0206] To a stirred solution of methyl (R, E)-1-(3-(1-methyl-1H-pyrazol-4-yl)acryloyl)piperidine-3-carboxylate-2,2,3,6,6-d5 19e (120 mg, 0.425 mmol) in THF (5 mL) was added Lithium aluminum hydride (17.74 mg, 0.468 mmol) at room temperature. The resulting mixture was stirred for 2 h. The reaction progress was monitored by TLC. The reaction was quenched by the addition of Na2SO4.10H2O (100 mg) at room temperature. The mixture was filtered; the filter cake was washed with EA (3 x 20 mL). The filtrate was concentrated under reduced pressure to give (R, E)-1-(3-(hydroxymethyl)piperidin-1-yl-2,2,3,6,6-d5)-3-(1-methyl-1H-pyrazol-4-yl)prop-2-en-1-one 19f (40 mg, 37.00%) as a white solid. LC-MS (ESI): m / z [M+H]+= 255.25.Step 6:

[0207] To a solution of (R, E)-1-(3-(hydroxymethyl)piperidin-1-yl-2,2,3,6,6-d5)-3-(1-methyl-1H-pyrazol-4-yl)prop-2-en-1-one 19f (40 mg, 0.079 mmol) in toluene (2 mL) was treated withAttorney Reference: ALLN-001WODPPA (30.30 mg, 0.111 mmol) at room temperature under a nitrogen atmosphere, followed by the addition of DBU (16.76 mg, 0.111 mmol). The resulting mixture was stirred at 110°C for 2 h. The mixture was concentrated under reduced pressure. The residue was dissolved in water (5 mL) and extracted with EA (3 x 10 mL). The combined organic layers were washed with brine (2 x 30 mL), dried over anhydrous Na2SO4and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2Cl2 / MeOH (9:1) to afford (R,E)-1-(3-(azidomethyl)piperidin-1-yl-2,2,3,6,6-d5)-3-(1-methyl-1H-pyrazol-4-yl)prop-2-en-1-one 19g (35 mg, 79.67%) as a yellow oil. LC-MS (ESI): m / z [M+H]+= 280.20.Step 7:

[0208] To a solution of (R, E)-1-(3-(azidomethyl)piperidin-1-yl-2,2,3,6,6-d5)-3-(1-methyl-1H-pyrazol-4-yl)prop-2-en-1-one 19g (35 mg, 0.125 mmol) in t-BuOH (2.5 mL) and H2O (1.5 mL) was treated with tert-butyl N-(prop-2-yn-l-yl)carbamate (25.28 mg, 0.163 mmol) at room temperature, followed by the addition of CuSO4.5H2O (6.26 mg, 0.025 mmol). To the above mixture was added Sodium ascorbate (4.96 mg, 0.025 mmol). The resulting mixture was stirred at room temperature for 2 h. H2O (10 mL) was added. The aqueous mixture was extracted with EA (3 x 10 mL), and the combined extracts were dried over anhydrous Na2SO4, concentrated under reduced pressure. The residue was purified by reversed-phase flash column chromatography with the following conditions: column, Cl 8 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 0% to 50% gradient in 10 min; detector, UV 254 nm. Removal of solvents gave tert-butyl (R, E)-((1-((1-(3-(1-methyl-1H-pyrazol-4-yl)acryloyl)piperidin-3-yl-2,2,3,6,6-d5)methyl)-1H-1,2,3-triazol-4-yl)methyl)carbamate 19h (35 mg, 64.29%) as a yellow oil. LC-MS (ESI): m / z [M+H]+= 435.25.Step 8:

[0209] To a solution of tert-butyl (R, E)-((1-((1-(3-(1-methyl-1H-pyrazol-4-yl)acryloyl)piperidin-3-yl-2,2,3,6,6-d5)methyl)-1H-1,2,3-triazol-4-yl)methyl)carbamate 19h (20 mg, 0.046 mmol) in 1,4-dioxane (2 mL) was added HCl in 1,4-dioxane (2.0 M, 2 mL) at room temperature. The resulting mixture was stirred for 2 h and concentrated under reduced pressure. The residue was basified to pH 8 with saturated NaHCO3(aq.) and purified by reversed-phase flash column chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. After lyophilization, the titled product (R,E)-1-(3-((4-(aminomethyl)-1H-1,2,3-triazol-1-yl)methyl)piperidin-1-yl-2,2,3,6,6-d5)-3-(1-methyl-1H-pyrazol-4-yl)prop-2-en-1-one Ex.19 (6.2 mg, 40.28%) was obtained as a white solid.1HNMR (300 MHz, CD3OD) 3 (ppm): 7.90 (s, 1H), 7.82 (s, 2H), 7.46 (d, I = 15.3 Hz, 1H), 6.78 - 7.02 (m, 1H), 4.30 - 4.51 (m, 2H), 3.75 - 4.02 (m, 5H), 1.75 - 1.89 (m, 2H), 1.31 - 1.60 (m, 2H); LC-MS (ESI): m / z [M+H]+= 335.25.Attorney Reference: ALLN-001WOExample 20: (R, E)-N-((1-((1-(3-(1-methyl-1H-pyrazol-4-yl)acryloyl)piperidin-3-yl)methyl)-1H-1,2,3-triazol-4-yl)methyl-d2)acetamide

[0210] To a stirred solution of (2E)-1-[(3R)-3-({4-[amino(2H2)methyl]-1,2,3-triazol-1-yl}methyl)piperidin-1-yl]-3-(1-methylpyrazol-4-yl)prop-2-en-1-one Ex.6 (50 mg, 0.151 mmol, available from Preparative Example 6) in pyridine (2 mL) was added acetyl chloride (14.21 mg, 0.181 mmol) dropwise at room temperature under a nitrogen atmosphere. The resulting mixture was stirred for 2 h. The reaction progress was monitored by LCMS. The reaction was quenched with ice water. The aqueous mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with water (3 x 30 mL), dried over anhydrous Na2SO4and concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, Cl 8 silica gel; mobile phase, MeCN in Water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min; detector, UV 254 nm. After lyophilization, the titled product N-[(l-{ [(3R)-l-[(2E)-3-(l-methylpyrazol-4-yl)prop-2-enoyl]piperidin-3-yl]methyl}-l,2,3-triazol-4-yl)(2H2)methyl]acetamide Ex.20 (20.3 mg, 36.03%) was obtained as a white solid.1HNMR (400 MHz, CD3OD) 5 (ppm): 7.88 (s, 1H), 7.86 (s, 1H), 7.79 (s, 1H), 7.42 - 7.45 (m, 1H), 6.74 - 6.91 (m, 1H), 4.30 - 4.50 (m, 2H), 4.10 - 4.25 (m, 1H), 3.95 - 4.09 (m, 1H), 3.89 (s, 3H), 3.20 - 3.30 (m, 1H), 2.80 - 3.09 (m, 1H), 2.10 - 2.30 (in, 1H), 1.95 (s, 3H), 1.75 - 1.90 (m, 2H), 1.44 - 1.62 (m, 2H); LC-MS (ESI): m / z [M+H]+= 374.20.Example 21: (R,E)-N-((1-((1-(3-(1-methyl-1H-pyrazol-4-yl)acryloyl)piperidin-3-yl-2,2,6,6-d4)methyl)-1H-1,2,3-triazol-4-yl)methyl)acetamide

[0211] Follow the procedure set forth in Preparative Example 20, by using (2£)-l-[(3S)-3-{[4-(aminomethyl)-l,2,3-triazol-l-yl]methyl}(2,2,6,6-2H4)piperidin-l-yl]-3-(l-methylpyrazol-4-yl)prop-2-en-l-one Ex.3 (210 mg, 0.630 mmol) prepared in Preparative Example 3, acetyl chloride (59.33 mg, 0.756 mmol), and pyridine (3 mL), the titled product N-[(1-{[(3S)-1-[(2E)-3-(1-methylpyrazol-4-yl)prop-2-enoyl](2,2,6,6-2H4)piperidin-3-yl]methyl}-1,2,3-triazol-4-Attorney Reference: ALLN-001WOyl)methyl] acetamide Ex.21 (26.4 mg, 11.16%) was prepared as a white solid.1HNMR (300 MHz, CD3OD) 8 (ppm): 7.88 (s, 2H), 7.79 (s, 1H), 7.44 (d, J= 15.3 Hz, 1H), 6.65 - 6.98 (m, 1H), 4.25 - 4.52 (m, 4H), 3.82 - 3.99 (m, 3H), 2.06 - 2.24 (m, 1H), 1.90 - 2.03 (m, 3H), 1.65 -1.89 (m, 2H), 1.29 - 1.60 (m, 2H); LC-MS (ESI): m / z [M+H]+= 376.20.Example 22: (R, E)-N-((l-((l-(3-(l-methyl-lH-pyrazol-4-yl)acryloyl)piperidin-3-yl-2, 2,6,6-d4)methyl)-lH-l,2,3-triazol-4-yl)methyl-d2)acetamide

[0212] Follow the procedure set forth in Preparative Example 20, by using (2E)-l-[(3S)-3-( { 4- [amino(2H2)methyl]- 1,2,3-triazol- 1 -yl }methyl)(2,2,6,6-2H4)piperidin- 1 -y 1] -3 -( 1 -methylpyrazol-4-yl)prop-2-en-l-one Ex.15 (50 mg, 0.149 mmol) prepared in Preparative Example 15, acetyl chloride (14.04 mg, 0.179 mmol), and pyridine (1 mL), the titled product N-[(l-{[(3S)-l-[(2E)-3-(l-methylpyrazol-4-yl)prop-2-enoyl](2,2,6,6-2H4)piperidin-3-yl]methyl}-l,2,3-triazol-4-yl)(2H2)methyl]acetamide Ex.22 (41.5 mg, 73.76%) was obtained as a white solid. ‘I INMR (300 MHz, CD3OD) 5 (ppm): 7.91 (s, 2H), 7.82 (s, 1H), 7.47 (d, 7= 15.0 Hz, 1H), 6.76 - 6.94 (m, 1H), 4.26 - 4.51 (m, 2H), 3.80 - 4.00 (s, 3H), 2.06 - 2.20 (m, 1H), 1.92 -2.05 (s, 3H), 1.69 - 1.91 (m, 2H), 1.25 - 1.65 (m, 2H); LC-MS (ESI): m / z [M+H]+= 378.20. Example 23: Evaluation of the pharmacokinetic properties of the exemplified compounds in rodents

[0213] Test#l in mice.

[0214] General procedure for mice: compound (30 mg / kg) is formulated in 0.25% methyl cellulose (4000cps) in purified water at a concentration of 6.0 mg / mL, well stirred, dosed to three male C57BL6 mice via oral gavage. Plasma samples were collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, and 8 hour. Approximately 0.3 mL blood sample was collected at each time point. Blood of each sample was transferred into plastic micro centrifuge tubes containing Heparin-Na. Collection tubes with blood samples and anticoagulant were inverted several times for proper mixing of the tube contents and then placed on wet ice prior to centrifugation for plasma. The blood samples were centrifugated at 4000g for 5 minutes at 4 °C to obtain plasma. All samples were stored in a freezer at -75 ± 15 °C prior to analysis. The concentration of the compound (analyte) was determined by LC / MS / MS. The pharmacokinetic (PK) parameters wereAttorney Reference: ALLN-001WOsummarized in the table below. AUC is an abbreviation for “area under the curve”.AUC(last) / D is dose normalized AUC.

[0215] Test #2 in rats.

[0216] General procedures: a compound (30 mg / kg) is formulated in 0.25% methyl cellulose (4000cps) in purified water at a concentration of 6.0 mg / mL, well stirred, dosed to three male SD rats via oral gavage. Plasma samples were collected at 0.033, 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hour. Approximately 0.2 mL blood sample was collected at each time point. Blood of each sample was transferred into plastic micro centrifuge tubes containing Heparin-Na.Collection tubes with blood samples and anticoagulant were inverted several times for proper mixing of the tube contents and then placed on wet ice prior to centrifugation for plasma. The blood samples were centrifugated at 4000g for 5 minutes at 4°C to obtain plasma. All samples were stored in a freezer at -75±15°C prior to analysis. The concentration of the compound (analyte) was determined by LC / MS / MS. The pharmacokinetic (PK) parameters were summarized in the table below. AUC is an abbreviation for “area under the curve”.AUC(last) / D is dose normalized AUC.

[0217] Attorney Reference: ALLN-001WOExample 24: Evaluation of the pharmacokinetic properties of the exemplified compounds in non-rodentsPharmacokinetic properties of compounds upon oral administration in dogs.

[0218] General procedures: a compound (10 mg / kg) was formulated in 0.25% methyl cellulose (4000cps) in purified water at a concentration of 2.0 mg / mL, well stirred, dosed to three male beagle dogs via oral gavage. Plasma samples were collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hour. Approximately 0.6 mL blood sample was collected at each time point. Blood of each sample was transferred into plastic micro centrifuge tubes containing Potassium (K2)EDTA. Whole blood samples were gently inverted several times to ensure anticoagulation then placed on wet ice prior to centrifugation. Within 30 minutes of collection, the samples were centrifuged at approximately 2,000 x g (force) for 10 minutes at 2 to 8°C to obtain plasma. The resultant plasma samples were immediately separated divided into 2 aliquots (-150 pl. each). Samples were maintained on wet ice throughout processing and stored in a freezer at -75±15°C prior to analysis. The concentration of the compound (analyte) was determined by LC / MS / MS. The pharmacokinetic (PK) parameters are summarized in the table below.

[0219] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it is readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

[0220] Accordingly, the preceding merely illustrates the principles of the invention. It will be appreciated that those skilled in the art will be able to devise various arrangements which, although not explicitly described or shown herein, embody the principles of the invention and are included within its spirit and scope. Furthermore, all examples and conditional language recited herein are principally intended to aid the reader in understanding the principles of theAttorney Reference: ALLN-001WOinvention and the concepts contributed by the inventors to furthering the art, and are to be construed as being without limitation to such specifically recited examples and conditions. Moreover, all statements herein reciting principles, aspects, and embodiments of the invention as well as specific examples thereof, arc intended to encompass both structural and functional equivalents thereof. Additionally, it is intended that such equivalents include both currently known equivalents and equivalents developed in the future, i.e., any elements developed that perform the same function, regardless of structure. Moreover, nothing disclosed herein is intended to be dedicated to the public regardless of whether such disclosure is explicitly recited in the claims.

[0221] The scope of the present invention, therefore, is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of present invention is embodied by the appended claims. In the claims, 35 U. S. C. § 112(f) is expressly defined as being invoked for a limitation in the claim only when the exact phrase "means for" or the exact phrase "step for" is recited at the beginning of such limitation in the claim; if such exact phrase is not used in a limitation in the claim, then 35 U. S. C. § 112 (f) is not invoked.

Claims

Attorney Reference: ALLN-001WOCLAIMSWhat is claimed is:

1. A compound of Formula (I):or a pharmaceutically acceptable salt thereof,wherein:the compound is enriched in deuterium;R1is -[C(R11)2]n-NRaRa;R2is selected from H, Rb, and Rbsubstituted with one or more Ca-Cs cycloalkyl; each R11is independently selected from hydrogen and Ci-Ce alkyl;n is 1, 2, or 3;each Rais independently selected from hydrogen, Ci-Ce alkyl, Ci-Ce haloalkyl, and -C(O)Rb, or two Raare taken together with the nitrogen atom to which they are bonded to form a 5- to 8-membered cycloheteroalkyl which may optionally include one or more of the same or different additional heteroatoms and which may optionally be substituted with one or more of the same or different Rb, -C(O)Rb, or both;each Rbis independently selected from Ci-Ce alkyl, -(Ci-Ce alkyleneXCj-Cs cycloalkyl), Ci-C6haloalkyl, -(Ci-Ce haloalkyl)(Cj-Cs cycloalkyl), and Cj-Cs cycloalkyl.

2. The compound of claim 1, wherein R1is enriched in deuterium.

3. The compound of any one of claims 1-2, wherein R1is -[CY2]n-NRaRa; andeach Y is independently selected from H and D.

4. The compound of claim 3, wherein R1is -CY2-NRaRa.

5. The compound of any one of claims 3-4, wherein each Rais independently selected from Y, Ci-C6alkyl, and -C(O)Rb, wherein Rbis Ci-C6alkyl.

6. The compound of claim 5, wherein R1is -[CY2]n-NH2 or -[CY2]n-NH-C(0)(Rb).

7. The compound of claim 6, wherein R1is -[CY2]n-NH2or -[CY2]n-NY-C(O)(methyl).Attorney Reference: ALLN-001WO8. The compound of claim 7, wherein R1is -CD2NH2 or -CD2NC(O)(CH3).

9. The compound of claim 1, wherein R1is -CH2NH2or -CH2NC(O)(CH3).

10. The compound of any one of claims 1-9, wherein R2is enriched in deuterium.

11. The compound of any one of claims 1-10, wherein R2is Ci-Ce alkyl or-(Ci-Ce alkylene)(C3-Cs cycloalkyl).

12. The compound of claim 11, wherein R2is methyl or -(methylene)(cyclopropyl).

13. The compound of claim 12, wherein R2is -CD3.

14. The compound of any one of claims 1-13, wherein positions 3 and 4 are each -CD=.

15. The compound of any one of claims 1-14, wherein positions 5 and 9 are each -CD2-.

16. The compound of any one of claims 1-15, wherein position 8 is17. The compound of any one of claims 1-16, wherein position 10 is -CD2-.

18. The compound of any one of claims 1-17, wherein each D represents deuterium incorporation of 80% or more.

19. The compound of any one of claims 1-18, wherein positions 1, 2, 11, or a combination thereof are each -CH=.

20. The compound of any one of claims 1-19, wherein position 6, position 7, or a combination thereof are each -CH2-.

21. The compound of any one of claims 1-20, wherein the carbon-carbon double bond at positions 3 and 4 has a trans configuration.

22. A pharmaceutical composition comprising a compound of any one of claims 1-21.

23. The pharmaceutical composition of claim 22, further comprising a pharmaceutically acceptable carrier.

24. A method for inhibiting shared epitope (SE) - calreticulin (CRT) binding, comprising:Attorney Reference: ALLN-001WOcontacting the calreticulin with an effective amount of a compound according to any one of claims 1-21.

25. The method of claim 24, wherein the contacting occurs in vitro.

26. The method of claim 24, wherein the contacting occurs in vivo.

27. The method of any one of claims 24-26. wherein the calreticulin is in an osteoclast.

28. The method of claim 27, wherein the contacting inhibits osteoclastogenesis.

29. The method of claim 26, wherein the contacting comprises administering the compound to a subject.

30. A method of treating a patient for a condition, the method comprising:administering an effective amount of a compound of any one of claims 1-21 to the patient.

31. The method of claim 30, wherein the condition is mediated by shared epitope (SE) -calreticulin (CRT) binding.

32. The method of any one of claims 30-31, wherein the condition is an immunological disorder or inflammatory disorder.

33. The method of any one of claims 30-32, wherein the condition is selected from the group consisting of rheumatoid arthritis, multiple sclerosis (MS), systemic lupus erythematosus, psoriasis, rejection of transplanted organ or tissue, inflammatory bowel disease (IBD), uveitis, Parkinson's disease, insulin dependent diabetes, non-insulin dependent diabetes, fibrosis, and periodontitis.

34. The method of any one of claims 30-33, wherein the condition includes bone erosion, bone loss, or a combination thereof.

35. The method of any one of claims 30-34, wherein the condition is a neuroinfl ammatory disorder.

36. The method of any one of claims 30-35, wherein the condition comprises a hematopoietic neoplasm.Attorney Reference: ALLN-001WO37. The method of claim 36, wherein the hematopoietic neoplasm is a myeloproliferative disorder (MPN).

38. The method of claim 36, wherein the hematopoietic neoplasm is selected from leukemias and lymphomas.

39. The method of any one of claims 30-38, further comprising administering one or more antiinflammatory compounds, one or more anti-rheumatic compounds, or a combination thereof to the patient.

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