TW202417415A - Inhibitors of plasma kallikrein - Google Patents

Abstract

The present disclosure provides compounds of Formula (IA) and their pharmaceutical compositions:

Description

Plasma microangiogenic factor inhibitors

This application claims priority to U.S. Provisional Patent Application Nos. 63/393,427 (filed on July 29, 2022) and 63/460,204 (filed on April 18, 2023), which are incorporated in their entirety as if fully set forth herein.

Plasma kallikrein is a serine protease that circulates in the blood as prekallikrein (an inactive pro-protein) and participates in the surface-mediated defense system through activation of factor XII and high molecular weight kininogen (HK)-involved signaling. Elements of the kallikrein-kinin system (KKS) are involved in activities such as surface-mediated defense responses, regulation of blood flow, fibrin deposition, blood pressure, smooth muscle contractility, pain perception, electrolyte transport, and mediator release. See Donald H. Miller, Harry S. Margolius, Chapter 19 The kallikrein-kinin-kininogen system, Editor(s): E. Edward Bittar, Neville Bittar, Principles of Medical Biology, Elsevier, Volume 8, 1997, Pages 363-384.

The present disclosure provides small molecule inhibitors of plasma microangiotensin and methods of using the inhibitors to treat diseases. Therefore, in various embodiments, the present disclosure provides a compound of formula (IA) or a pharmaceutically acceptable salt thereof: (IA)

In formula (IA), ^D1 is N or ^CR1 , ^D2 is N or ^CR2 , ^D3 is N or ^CR3 , and ^D4 is N or ^CR4 . In addition, no more than three of ^D1 , ^D2 , ^D3 , and ^D4 are N at the same time.

The substituents R ¹ , R ² , R ³ , and R ⁴ are independently selected from the group consisting of H, C ₂ -C ₆ -alkenyl, C ₁ -C ₆ -halogenalkyl, halogen, NR ^a R ^b , OR ^a , -NR ^a C(O)R ^b , -C(O)R ^a , -C(O)halogen, -OC(O)R ^a , -OC(O)OR ^a , -C(O)OR ^a , C ₆ -C ₁₀ -aryl, CN, -S(O) _0-2 R ^a , -S(O) ₂ OR ^a , and NO ₂ .

Each of ^Ra and ^Rb is independently selected from the group consisting of H, _C1 - _C6 -alkyl, and _C1 - _C6 -haloalkyl.

The substituents R ^c1 , R ^c2 , and R ^c3 are independently selected from the group consisting of H, C ₁ -C ₆ -alkyl, and C ₁ -C ₆ -haloalkyl.

Q ₁ , Q ₂ , and Q ₃ are independently selected from the group consisting of CR ⁵ , N, O, and S.

R ⁵ is selected from the group consisting of H, C ₁ -C ₆ -alkyl, OR ^a , —(C ₁ -C ₆ -alkyl)OR ^a , and C ₃ -C ₁₀ -cycloalkyl.

The ring member P is C or N.

L ¹ is -SO ₂ - or -C ₁ -C ₈ -alkylene-.

Moiety is a divalent monocyclic or bicyclic moiety selected from the group consisting of C ₃ -C ₁₀ -cycloalkyl, C ₆ -C ₁₀ -aryl, 3 to 10 membered heterocycloalkyl (wherein 1 to 4 ring members are independently selected from N, O, and S), 5 to 10 membered heteroaryl (wherein 1 to 4 heteroaryl members are independently selected from N, O, and S), and fused combinations thereof.

^L2 is selected from the group consisting of a bond, _-C1 - _C8 -alkylene, _-C2 - _C8 -alkenylene, _-C2 - _C8 -alkynylene, and a divalent moiety selected from the group consisting of _C3 - _C10 -cycloalkyl, _C3 - _C10 -cycloalkenyl, _C6 - _C10 -aryl, 5- to 10-membered heteroaryl (wherein 1 to 4 heteroaryl members are independently selected from N, O, and S), ( _C1 - _C8 -alkyl) _C3 - _C10 -cycloalkyl, ( _C1 - _C8 -alkyl) _C3 - _C10 -cycloalkenyl, ( _C1 - _C8 -alkyl) _C6 - _C10 -aryl, and ( _C1 - _C8 -alkyl) 5 to 10 membered heteroaryl (wherein 1 to 4 heteroaryl members are independently selected from N, O, and S). In L ² , any cycloalkyl, cycloalkenyl, aryl, and heteroaryl is monocyclic or bicyclic. L ² is optionally substituted with 1 to 3 substituents selected from the group consisting of C ₁ -C ₆ -alkyl, C ₁ -C ₆ -haloalkyl, and CN.

Z is selected from the group consisting of ^-ORc , -OC(O) ^Rc , -OC(O) ^{NRcRd ,} -S(O) _0-2Rc , ^-CN , -C(O) ^Rc , ^-C (O)ORc, -C(O) ^{NRcRd ,} -C(S) ^Rc _, ^-NRcRd , =NRc, ^{-NRcC (} O) ^{NRcRd , -NRcCO2Rd} , ^-NRc -NO, _-NO2 , ^-NRc - ^ORd , -N=C=O, -N= ^C =S ^, and ^{-NRc - NRcRd} ; and

Examples of R ^c and R ^d are independently selected from H, C ₁ -C ₆ -alkyl, C ₁ -C ₆ -haloalkyl, and C ₆ -C ₁₀ -aryl.

In another embodiment, the compound is one of Formula I: (Formula I)

In Formula I, the moiety R is i) a cyclic hydrocarbon, bicyclic hydrocarbon, or heterocyclic ring containing up to 10 atoms consisting of C or N, or R is ii) selected from the group consisting of cyclopentyl, cyclohexyl, phenyl, naphthyl, pyridine, pyrimidine, indene, 2,3-dihydro- 1H -indene, or any saturated and unsaturated cyclic hydrocarbon or heterocyclic ring thereof.

Z is selected from the group consisting of -OH, -OR', -OC(O)H, -OC(O)R', -OC(O) _NH2 , -OC(O)NHR', -OC(O)NH(R') ₂ , -SH, -SR', -S(O)R', -S(O) _2R ', -CN, -C(O)H, -C(O)R', -C(O)OH, -C(O)OR', -C(O) _NH2 , -C(O)NHR', _-C (O)NH(R') ₂ , -C(S)R', _-NH2 , _-NH2R ', -NR'2, =NH, =NR', -NHC(O)NH2, -NR'C(O) _NH2 , -NR'C(O)NHR', -NR'C(O) _{N(R')2 , -NHCO2} H, -NHCO ₂ R', -NR'CO ₂ R', -NH-NO, -NR'-NO, -NO ₂ , -NH-OH, -OH, -NR'-OR', -N=C=O, -N=C=S, -NH-NH 2 , and -NH-NHR', wherein each R' is independently an alkyl or alkyl halide. In some embodiments, Z is not a halogen or hydrogen.

Q ₁ , Q ₂ , and Q ₃ are selected from the group consisting of C, N, O, or S.

P is selected from the group consisting of C or N.

_L1 is a linking group selected from the group consisting of _C1 - _C8 alkyl linkers or _SO2 .

L ₂ is selected from i) a hydrocarbon without a double bond to O or S, ii) a hydrocarbon without a heteroatom such as O, N, or S; or iii) a linker selected from the group consisting of: a C ₁ -C ₈ alkyl linker comprising a saturated hydrocarbon, an unsaturated hydrocarbon, a branched hydrocarbon, a cyclic hydrocarbon, and a combination thereof; a cyclic hydrocarbon selected from the group consisting of: cyclopentyl, cyclohexyl, phenyl, naphthyl, indene, 2,3-dihydro-1 H -indene, or any saturated or unsaturated cyclic hydrocarbon thereof. In some embodiments, L ₂ is a hydrocarbon containing a heteroatom such as O, N, or S.

In some embodiments, the compositions comprise a compound of Formula I and a pharmaceutically acceptable salt thereof.

The present disclosure provides compounds, compositions, and methods for inhibiting plasma microangiotensin ("PKal"). The compounds can be used in exemplary embodiments to treat inflammatory and ocular diseases. Definitions

Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which the present disclosure pertains.

As used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context dictates otherwise. Thus, for example, reference to "an agent" includes a plurality of such agents and reference to "the cell" includes one or more cells (or cells) and equivalents thereof known to those skilled in the art, and so forth.

When ranges are used herein for physical properties (such as molecular weight) or chemical properties (such as chemical formula), all combinations and subcombinations of ranges and specific embodiments therein are intended to be included. When referring to a number or a numerical range, the term "about" means that the referenced number or numerical range is approximate within the experimental variability (or within statistical experimental error), so that the number or numerical range (in some cases) will vary between 1% and 15% of the stated number or numerical range.

In the present disclosure, the number of atoms of a particular element in a substituent is generally given as a range, such as an alkyl group containing 1 to 4 carbon atoms or a _C1-4 alkyl group. Reference to such ranges is intended to include specific reference to each of the integer numbers of atoms within the specified range. For example, an alkyl group of 1 to 4 carbon atoms includes each of _C1 , _C2 , _C3 , and _C4 . For example, a _C1-12 heteroalkyl group includes 1 to 12 carbon atoms in addition to one or more heteroatoms. Other numbers of atoms and other types of atoms may be indicated in a similar manner.

The term "comprising" (and related terms such as "comprise" or "comprises" or "having" or "including") is not intended to exclude certain other embodiments, such as any composition of matter, composition, method, or process, or the like (described herein), "consist of" or "consist essentially of".

"Alkyl" refers to a straight or branched chain hydrocarbon group comprising 1 to about 20 carbon atoms. For example, an alkyl group can have 1 to 10 carbon atoms or 1 to 6 carbon atoms. Exemplary alkyl groups include straight chain alkyl groups such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, and the _like , and also include branched chain isomers of straight chain alkyl groups such as, but not limited to, -CH( _{CH3 ) 2} , -CH( _{CH3 )} ( _CH2CH3 ), _-CH ( _CH2CH3 ) ₂ , -C( _{CH3 ) 3} , -C( _CH2CH3 ) ₃ , -CH2CH( _CH3 ) _{2 ,} -CH2CH _{( CH3} )( _{CH2CH3 )} , _{-CH2CH ( CH2CH3} ) ₂ , _-CH2C ( _CH3 ) ₃ , -CH2C( _CH2CH3 ) ₃ , -CH( _CH3 )CH( _CH3 )(CH2CH ₃ ), -CH ₂ CH ₂ CH(CH ₃ ) ₂ , -CH ₂ CH ₂ CH(CH ₃ )(CH ₂ CH ₃ ), -CH ₂ CH ₂ CH(CH ₂ CH ₃ ) ₂ , -CH ₂ CH ₂ C(CH ₃ ) ₃ , -CH ₂ CH ₂ C(CH ₂ CH ₃ ) ₃ , -CH(CH ₃ )CH ₂ CH(CH ₃ ) ₂ , -CH(CH ₃ )CH(CH ₃ ) CH(CH ₃ ) ₂ , and the like. Thus, alkyl includes primary alkyl, secondary alkyl, and tertiary alkyl. Alkyl groups may be substituted with one or more substituents as described herein, such as, for example, one or more halogens.

The terms "halogen", "halide", and "halo" refer to -F or fluoro, -Cl or chloro, -Br or bromo, or -I or iodo.

The term "alkenyl" refers to a straight or branched chain hydroxyl group comprising 2 to about 20 carbon atoms, having 1 to 3, 1 to 2, or at least one carbon to carbon double bond. Alkenyl groups may be unsubstituted or optionally substituted with one or more substituents as described herein.

"Alkyne" or "alkyne" refers to a straight or branched unsaturated hydrocarbon having the indicated number of carbon atoms and at least one triple bond. Examples of ( _C2 - _C8 )alkynyl groups include, but are not limited to, acetylene, propyne, 1-butyne, 2-butyne, 1-pentyne, 2-pentyne, 1-hexyne, 2-hexyne, 3-hexyne, 1-heptyne, 2-heptyne, 3-heptyne, 1-octyne, 2-octyne, 3-octyne, and 4-octyne. Alkyne groups may be unsubstituted or optionally substituted with one or more substituents as described herein.

The term "cycloalkyl" refers to a saturated monocyclic, bicyclic, tricyclic, or polycyclic, 3 to 14 membered ring system, such as C ₃ -C ₈ -cycloalkyl. The cycloalkyl group may be attached via any atom. Representative examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. The cycloalkyl group may be unsubstituted or optionally substituted with one or more substituents as described herein.

Embodiments of plasma microangiogenic factor inhibitors include compounds containing C ₁ -C ₈ alkyl linkers. In embodiments, "C _1-8 alkyl" is characterized by a branched or unbranched alkyl group having 1 to 8 carbon atoms. C _1-8 alkyl includes but is not limited to methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclopropylmethyl, n-butyl, isobutyl, dibutyl, tertiary butyl, cyclobutyl, pentyl, and cyclopentyl.

"Aryl" (Ar) when used alone or as part of another term means a carbocyclic aromatic group (whether or not fused) having the number of carbon atoms specified, or if no number is specified, up to 14 carbon atoms, such as _C6 - _C10 -aryl or _C6 - _C14 -aryl. In embodiments, Ar may be characterized as an aromatic group having a ring system comprising carbon atoms with conjugated π electrons (e.g., phenyl). The term includes aryl groups having 6 to 12 carbon atoms. Aryl groups may optionally include monocyclic, bicyclic, or tricyclic rings, wherein each ring has five or six members. Examples of aryl groups include phenyl, naphthyl, biphenyl, phenanthrenyl, fused tetraphenyl, and the like (see, e.g., Lang's Handbook of Chemistry (Dean, JA, ed) ^13th ed. Table 7-2 [1985]). "Aryl" also contemplates an aryl ring that is part of a fused polycyclic system, such as an aryl group fused to a cycloalkyl group as defined herein. An exemplary aryl group is phenyl. Aryl groups may be unsubstituted or optionally substituted with one or more substituents as described herein.

The term "heteroatom" refers to N, O, and S. The disclosed compounds containing N or S atoms can be optionally oxidized to the corresponding N-oxide, sulfide, or sulfonate compounds.

"Heteroaryl" (alone or in combination with any other moiety described herein) is a monocyclic aromatic ring structure containing 5 to 10 (e.g., 5 or 6) ring atoms, or a bicyclic aromatic group having 8 to 10 atoms, which contains one or more (e.g., 1 to 4, 1 to 3, or 1 to 2) heteroatoms independently selected from the group consisting of O, S, and N. Heteroaryl is also intended to include oxidized S or N, such as sulfinyl, sulfonyl, and N-oxides of tertiary ring nitrogens. The carbon or heteroatom is the point of attachment to the heteroaryl ring structure, thereby producing a stable compound. Examples of heteroaryl groups include, but are not limited to, pyridyl, pyridazinyl, pyrazinyl, quinaoxalyl, indolizinyl, benzo[b]thienyl, quinazolinyl, purinyl, indolyl, quinolinyl, pyrimidinyl, pyrrolyl, pyrazolyl, The heteroaryl group may be unsubstituted or optionally substituted with one or more substituents as described herein.

"Heterocycloalkyl" is a saturated or partially unsaturated nonaromatic monocyclic, bicyclic, tricyclic, or polycyclic ring system having 3 to 14 (e.g., 3 to 6) atoms, wherein 1 to 3 carbon atoms in the ring are heteroatoms of O, S, or N. Ring heteroatoms may also be oxidized S or N, such as sulfinyl, sulfonyl, and N-oxides of tertiary ring nitrogens. Heterocycloalkyl may be fused to another ring system, such as an aryl or heteroaryl with 5 to 6 ring members. The point of attachment of the heterocycloalkyl ring is at a carbon or heteroatom, thereby maintaining a stable ring. Examples of heterocycloalkyl include, but are not limited to, N-morpholino, tetrahydrofuranyl, dihydropyridinyl, piperidinyl, pyrrolidinyl, piperazinyl, dihydrobenzofuranyl, and dihydroindolyl. Heterocycloalkyl may be unsubstituted or optionally substituted with one or more substituents as described herein.

The terms "nitrile" or "cyano" are used interchangeably and refer to a -CN group.

The compounds described herein may exist in various isomeric forms, including configurational, geometrical, and conformational isomers, including, for example, cis or trans configurations. The compounds may also exist in one or more tautomeric forms, including a single tautomeric isomer and a mixture of multiple tautomeric isomers. The term "isomer" is intended to encompass all isomeric forms of the disclosed compounds, including tautomeric isomers of the compounds. The disclosed compounds may also exist in open chain or cyclized forms. In some cases, one or more of the cyclized forms may be due to the loss of water. The specific composition of the open chain and cyclized forms may depend on how the compound is isolated, stored, or administered. For example, a compound may exist primarily in an open chain form under acidic conditions but cyclize under neutral conditions. All forms are included in this disclosure.

Some of the compounds described herein may have asymmetric centers and therefore exist as different mirror isomers and non-mirror isomers. The compounds described herein may be in the form of optical isomers or non-mirror isomers. Therefore, the present disclosure covers the compounds described herein and their uses in the form of their optical isomers, non-mirror isomers, and mixtures thereof (including racemic mixtures). Optical isomers of the disclosed compounds can be obtained by known techniques, such as asymmetric synthesis, chiral chromatography, simulated moving bed technology, or by chemical separation of stereoisomers (by using optically active identifying separating agents).

Unless otherwise indicated, the term "stereoisomer" means one stereoisomer of a compound that is substantially free of other stereoisomers of the compound. Thus, a stereoisomerically pure compound having one chiral center will be substantially free of the opposite mirror image isomer of the compound. A stereoisomerically pure compound having two chiral centers will be substantially free of other non-mirror image isomers of the compound. A typical stereoisomerically pure compound comprises greater than about 80% by weight of one stereoisomer of the compound and less than about 20% by weight of other stereoisomers of the compound, e.g., greater than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of other stereoisomers of the compound, or greater than about 95% by weight of one stereoisomer of the compound and less than about 5% by weight of other stereoisomers of the compound, or greater than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of other stereoisomers of the compound, or greater than about 99% by weight of one stereoisomer of the compound and less than about 1% by weight of other stereoisomers of the compound. The stereoisomers described above may be considered as compositions comprising both stereoisomers, which are present in their respective percentages as described herein.

If there is a discrepancy between a drawn structure and the name given to that structure, the drawn structure controls. In addition, if the stereochemistry of a structure or portion of a structure is not indicated by bold or dashed lines, the structure or portion of a structure is to be interpreted as encompassing all of its stereoisomers. However, in some cases, if more than one chiral center is present, the structure and name may be indicated as a single mirror image isomer to help describe the relative stereochemistry. One of ordinary skill in the art will know if a compound is prepared as a single mirror image isomer from the process used to prepare it.

As used herein, and unless otherwise specified to the contrary, the term "compound" is inclusive, that is, it encompasses a compound or its pharmaceutically acceptable salts, stereoisomers, isotope-like molecules, and/or tautomers. Thus, for example, a compound includes a pharmaceutically acceptable salt of a tautomer of the compound. Similarly, a compound includes a pharmaceutically acceptable salt of an isotope-like molecule of the compound.

In the present disclosure, a "pharmaceutically acceptable salt" is a pharmaceutically acceptable, organic or inorganic acid or alkaline salt of a compound described herein. Representative pharmaceutically acceptable salts include, for example, alkali metal salts, alkali earth salts, ammonium salts, water-soluble and water-insoluble salts such as acetate, ansorlate (4,4-diaminostilbene-2,2-disulfonate), benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium, calcium edetate, camphorsulfonate, carbonate, chloride, citrate, clavulariate, dihydrochloride, edetate, edisylate, estolide sulfate, and succinate. late), esylate, fiunarate, gluceptate, gluconate, glutamine, glycollylarsanilate, hexafluorophosphate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate oate), iodide, 2-hydroxyethanesulfonate (isothionate), lactate, lactobionate, laurate, apple acid salt, cis-butenedioate, mandelate, methanesulfonate, methyl bromide, methyl nitrate, methyl sulfate, mucate, naphthalenesulfonate (napsylate), nitrate, N-methylglucamine (N-methylglucamine) ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate, palmitic acid salt, palmitic acid salt (1,1-methylene-bis- The pharmaceutically acceptable salts include 2-hydroxy-3-naphthoate, einbonate), pantothenate, phosphate/diphosphate, picrate, polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate, subacetate, succinate, sulfate, sulfosalicylate, suramate, tannate, tartrate, teoclate, toluenesulfonate, triethiodide, and valerate. A pharmaceutically acceptable salt may have more than one charged atom in its structure. In this case, the pharmaceutically acceptable salt may have multiple relative ions. Thus, a pharmaceutically acceptable salt may have one or more charged atoms and/or one or more counter ions.

The terms "treat", "treating" and "treatment" refer to the improvement or elimination of a disease or symptoms associated with a disease. In various embodiments, these terms refer to minimizing or slowing the spread, progression, or worsening of a disease as a result of administering one or more disease prevention or treatment compounds described herein to a patient suffering from such a disease.

The terms "prevent", "preventing" and "prevention" refer to preventing the onset, recurrence, or spread of a disease in a patient as a result of administering a compound described herein.

The term "effective amount" refers to an amount of a compound or other active ingredient as described herein that is sufficient to provide a therapeutic or disease preventive benefit in the treatment or prevention of a disease or to delay or minimize symptoms associated with the disease. In addition, with respect to a compound as described herein, a therapeutically effective amount means an amount of a therapeutic agent (alone or in combination with other therapies) that provides a therapeutic benefit in the treatment or prevention of a disease. Used in conjunction with a compound as described herein, the term may encompass an amount that enhances the overall therapy, reduces or avoids symptoms or causes of a disease, or increases the efficacy of a treatment or has a synergistic effect with another therapeutic agent.

"Patient" or "subject" includes animals, such as humans, cows, horses, sheep, lambs, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, rabbits, or guinea pigs. According to some embodiments, the animal is a mammal, such as non-primates and primates (e.g., monkeys and humans). In one embodiment, the patient is a human, such as a human infant, child, adolescent, or adult. In this disclosure, the terms "patient" and "subject" are used interchangeably. Inhibitors of Plasma Microangiotensin Compounds of Formula IA

In some embodiments, the inhibitor is a compound of formula (IA) or a pharmaceutically acceptable salt thereof: (IA)

In formula (IA), ^D1 is N or ^CR1 , ^D2 is N or ^CR2 , ^D3 is N or ^CR3 , and ^D4 is N or ^CR4 . In addition, no more than three of ^D1 , ^D2 , ^D3 , and ^D4 are N at the same time.

The substituents R ¹ , R ² , R ³ , and R ⁴ are independently selected from the group consisting of H, C ₂ -C ₆ -alkenyl, C ₁ -C ₆ -halogenalkyl, halogen, NR ^a R ^b , OR ^a , -NR ^a C(O)R ^b , -C(O)R ^a , -C(O)halogen, -OC(O)R ^a , -OC(O)OR ^a , -C(O)OR ^a , C ₆ -C ₁₀ -aryl, CN, -S(O) _0-2 R ^a , -S(O) ₂ OR ^a , and NO ₂ .

Each of ^Ra and ^Rb is independently selected from the group consisting of H, _C1 - _C6 -alkyl, and _C1 - _C6 -haloalkyl.

The substituents R ^c1 , R ^c2 , and R ^c3 are independently selected from the group consisting of H, C ₁ -C ₆ -alkyl, and C ₁ -C ₆ -haloalkyl.

Q ₁ , Q ₂ , and Q ₃ are independently selected from the group consisting of CR ⁵ , N, O, and S.

R ⁵ is selected from the group consisting of H, C ₁ -C ₆ -alkyl, OR ^a , —(C ₁ -C ₆ -alkyl)OR ^a , and C ₃ -C ₁₀ -cycloalkyl.

The ring member P is C or N.

L ¹ is -SO ₂ - or -C ₁ -C ₈ -alkylene-.

Part is a divalent monocyclic or bicyclic moiety selected from the group consisting of C ₃ -C ₁₀ -cycloalkyl, C ₆ -C ₁₀ -aryl, 3 to 10 membered heterocycloalkyl (wherein 1 to 4 ring members are independently selected from N, O, and S), 5 to 10 membered heteroaryl (wherein 1 to 4 heteroaryl members are independently selected from N, O, and S), and fused combinations thereof.

^L2 is selected from the group consisting of a bond, _-C1 - _C8 -alkylene, _-C2 - _C8 -alkenylene, _-C2 - _C8 -alkynylene, and a divalent moiety selected from the group consisting of _C3 - _C10 -cycloalkyl, _C3 - _C10 -cycloalkenyl, _C6 - _C10 -aryl, 5- to 10-membered heteroaryl (wherein 1 to 4 heteroaryl members are independently selected from N, O, and S), ( _C1 - _C8 -alkyl) _C3 - _C10 -cycloalkyl, ( _C1 - _C8 -alkyl) _C3 - _C10 -cycloalkenyl, ( _C1 - _C8 -alkyl) _C6 - _C10 -aryl, and ( _C1 - _C8 -alkyl) 5 to 10 membered heteroaryl (wherein 1 to 4 heteroaryl members are independently selected from N, O, and S). In L ² , any cycloalkyl, cycloalkenyl, aryl, and heteroaryl is monocyclic or bicyclic. L ² is optionally substituted with 1 to 3 substituents selected from the group consisting of C ₁ -C ₆ -alkyl, C ₁ -C ₆ -haloalkyl, and CN.

Z is selected from the group consisting of ^-ORc , -OC(O) ^Rc , -OC(O) ^{NRcRd ,} -S(O) _0-2Rc , ^-CN , -C(O) ^Rc , ^-C (O)ORc, -C(O) ^{NRcRd ,} -C(S) ^Rc _, ^-NRcRd , =NRc, ^{-NRcC (} O) ^{NRcRd , -NRcCO2Rd} , ^-NRc -NO, _-NO2 , ^-NRc - ^ORd , -N=C=O, -N= ^C =S ^, and ^{-NRc - NRcRd} ; and

Examples of R ^c and R ^d are independently selected from H, C ₁ -C ₆ -alkyl, C ₁ -C ₆ -haloalkyl, and C ₆ -C ₁₀ -aryl.

In some embodiments, three of ^D1 , ^D2 , ^D3 , and ^D4 are N. In other embodiments, two of ^D1 , ^D2 , ^D3 , and ^D4 are N. In still other embodiments, one of ^D1 , ^D2 , ^D3 , and ^D4 is N. In still additional embodiments, no more than one of ^D1 , ^D2 , ^D3 , and ^D4 is N. In illustrative embodiments, the ring containing ^D1 , ^D2 , ^D3 , and ^D4 is selected from the group consisting of: , ,and .

In certain embodiments, the ring system containing D ¹ , D ² , D ³ , and D ⁴ is: .

In various embodiments, R ¹ , R ² , R ³ , and R ⁴ are independently selected from the group consisting of H, OR ^a , halogen, and CN. In some embodiments, R ¹ , R ² , R ³ , and R ⁴ are independently selected from the group consisting of H and halogen. For example, in one embodiment, at least one of R ¹ , R ² , R ³ , and R ⁴ is halogen. In another embodiment, each of R ¹ , R ² , R ³ , and R ⁴ is H.

According to various embodiments, some compounds of Formula (IA) are those wherein each of R ^c1 , R ^c2 , and R ^c3 is H. Thus, for example, some embodiments provide a ring system containing D ¹ , D ² , D ³ , and D ⁴ : , ,or compound of.

In various embodiments, ^Q1 is C; each of ^Q2 and ^Q3 is independently selected from the group consisting of ^CR5 , N, O, and S; and P is selected from the group consisting of C and N. In one embodiment, P is N. In additional embodiments, ^Q2 is ^CR5 and ^Q3 is N, or ^Q2 is N and ^Q3 is ^CR5 . Illustrative embodiments include a ring system containing P, ^Q1 , ^Q2 , and ^Q3 : , , , ,and compound of.

In additional embodiments, L ¹ is -C ₁ -C ₈ -alkylene-, such as -C ₁ -C ₃ -alkylene-. In an illustrative embodiment, L ¹ is methylene.

In other embodiments, part is a divalent monocyclic C ₆ -C ₁₀ -aryl group. In an exemplary embodiment, Department .

In additional embodiments, the disclosure provides compounds of formula (IA), wherein ^L is selected from -C ₁ -C ₈ -alkylene, C ₆ -C ₁₀ -aryl, and -(C ₁ -C ₈ -alkyl)C ₆ -C ₁₀ -aryl, any of which is optionally substituted, as described herein. In some embodiments, L is -C ₁ -C ₈ -alkylene, such as -C ₁ -C ₃ -alkylene. As generally described herein, the alkyl (or alkylene) portion may be linear or branched. In other embodiments, ^L is C ⁶ _{-C 10} -aryl, including phenyl.

In some embodiments, the moiety Z is selected from the group consisting of -OR ^c , CN, -C(O)OR ^c , and -C(O)NR ^c R ^d . In various embodiments, each R ^c and R ^d is independently H or C ₁ -C ₆ -alkyl. Thus, for example, Z can be selected from the group consisting of OH, OCH ₃ , -COOH, -C(O)NH ₂ , and -C(O)NHCH ₃ . In another embodiment, Z is CN. Compounds of Formula I

In some embodiments, the present disclosure provides PKal inhibitors of Formula I. (I) wherein R is i) a cyclic hydrocarbon, bicyclic hydrocarbon, or heterocyclic ring containing up to 10 atoms consisting of C or N, or R is ii) selected from the group consisting of cyclopentyl, cyclohexyl, phenyl, naphthyl, pyridine, pyrimidine, indene, 2,3-dihydro- 1H -indene, or any saturated and unsaturated cyclic hydrocarbon or heterocyclic ring thereof.

In some embodiments, Z is selected from the group consisting of -OH, -OR', -OC(O)H, -OC(O)R', -OC(O) _NH2 , -OC(O)NHR', -OC(O)NH(R') ₂ , -SH, -SR', -S(O)R', -S(O) _2R ', -CN, -C(O)H, -C(O)R', -C(O)OH, -C(O)OR', -C(O) _NH2 , -C(O)NHR', -C(O)NH(R') ₂ , -C(S ₎ R', -NH2, -NH2R', -NR'2, =NH, =NR', -NHC( _O )NH2, -NR'C(O) _NH2 , _-NR'C (O)NHR', -NR'C(O) _N (R') _{2 In some embodiments ,} Z is not a halogen or hydrogen.

In some embodiments, Q ₁ , Q ₂ , and Q ₃ are selected from the group consisting of C, N, O, or S.

In some embodiments, P is selected from the group consisting of C or N.

In some embodiments, L ₁ is a linker selected from the group consisting of a C ₁ -C ₈ alkyl linker or SO ₂ .

In some embodiments, _L2 is selected from i) a hydrocarbon without a double bond to O or S, ii) a hydrocarbon without heteroatoms such as O, N, or S; or iii) a linker selected from the group consisting of: a _C1 - _C8 alkyl linker comprising a saturated hydrocarbon, an unsaturated hydrocarbon, a branched hydrocarbon, a cyclic hydrocarbon, and a combination thereof; a cyclic hydrocarbon selected from the group consisting of: cyclopentyl, cyclohexyl, phenyl, naphthyl, indene, 2,3-dihydro- 1H -indene, or any saturated or unsaturated cyclic hydrocarbon thereof. In some embodiments, _L2 is a hydrocarbon containing heteroatoms such as O, N, or S.

In some embodiments, the compositions comprise a compound of Formula I and a pharmaceutically acceptable salt thereof.

In some embodiments, the compound comprises a subgenus with the following conditions: _Q1 , _Q2 and _Q3 are selected from the group consisting of C and N, P is N, _L1 is _CH2 , R is para-substituted benzene, _L2 is a methylene or ethylene alkyl linker at the para position of the R group, and Z is selected from the group consisting of CN and OH.

Additional embodiments include the following: • Q ₁ is C, Q ₂ is C, Q ₃ is N, L ₂ is a methylene bond linker, and Z is CN; • Q ₁ is C, Q ₂ is C, Q ₃ is N, L ₂ is a methylene bond linker, and Z is OH; • Q ₁ is C, Q ₂ is C, Q ₃ is N, L ₂ is an ethyl bond linker and Z is CN; • Q ₁ is C, Q ₂ is N, Q 3 is C, L 2 is an ethyl bond linker and Z is CN; • Q 1 is C, Q 2 is N, Q ₃ is C, L ₂ is an ethyl bond linker and Z is CN; • Q ₁ , Q ₂ , and Q ₃ are selected from the group consisting of C or N, P is N, L ₁ is CH2, R is benzene, L2 is selected from the group consisting of a branched ethyl bond linker and an isopropyl bond linker at the para position of the R group and Z is a nitrile group (CN); • Q • Q ₁ is C, Q ₂ is C, Q ₃ is N, and L ₂ is a branched ethyl linker; • Q ₁ is C, Q ₂ is N, Q ₃ is C, and L ₂ is a branched ethyl linker; • Q ₁ is C, Q ₂ is C, Q ₃ is N, and L ₂ is an isopropyl linker; • Q ₁ is C, Q ₂ is N, Q ₃ is C, and L ₂ is an isopropyl linker; • Q ₁ is C, Q ₂ is selected from the group consisting of N, O, or S, Q ₃ is selected from the group consisting of C or N, L ₁ is a methylene or ethyl linker, R is benzene, L ₂ is a methylene linker at the para position of the R group, and Z is CN; • Q ₂ is N, Q ₃ is C, P is N, and L • Q ₁ is a methylene bond linker; • Q ₂ is O, Q ₃ is N, P is C, and L ₁ is a methylene bond linker; • Q ₂ is S, Q ₃ is N, P is C, and L ₁ is a methylene bond linker; • Q ₂ is C, Q ₃ is N, P is N, and L ₁ is an ethylene bond linker; • Q ₁ is C, Q ₂ is C, Q ₃ is N, P is N, L ₁ is CH ₂ , R is benzene, L ₂ is a phenyl bond linker in the para position of the R group, and Z is CN; • Q ₁ is C, Q ₂ is C, Q ₃ is N, P is N, L ₁ is CH ₂ , R is benzene, L ₂ is a benzyl bond linker in the para position of the R group, and Z is CN in the para position; • Q ₁ is C, Q ₂ is C, Q ₃ is N, P is N, L • Q 1 is C, Q 2 is C, Q 3 is N, P is N, _{L 1} is CH ₂ , R is benzene, L ₂ is a methylene or ethylidene bond linker in the para position of the R group, and Z is selected from the group consisting of methoxy, carboxylic acid, amide, and C1 alkyl (methyl) substituted amide bonded to N; • Q ₁ is C, Q ₂ is C, Q ₃ is N, P is N, L ₁ is CH ₂ , R is benzene, L ₂ is an ethylidene bond linker in the para position of the R group, and Z is methoxy; • Q ₁ is C, Q ₂ is C, Q ₃ is N, P is N, L ₁ is CH 2 , R is benzene, L 2 is a methylene bond linker in the para position of the R group, and Z is carboxylic acid; • Q ₁ is C, Q ₂ is C, Q ₃ is N, P is N, L ₁ is CH ₂ , R is benzene, L ₂ is a methylene bond linker in the para position of the R group, and Z is carboxylic acid. • _Q1 is C, Q2 is C, _Q3 is N, P is N, _L1 is CH2, R is benzene, _L2 is a methylene bond linker in the para position of the R group, and Z is an amide; • Q1 is C, _Q2 is C, Q3 is N, P is N, L1 is CH2, R is benzene, _L2 is a methylene bond linker in the para position of the R group, and Z is an amide substituted by a C1 alkyl(methyl) bond to N; • _Q1 is C, _Q2 is C, _Q3 is N, P is N, _L1 is _CH2 , R is benzene, L2 is a cyclopentylene or a methylene bond linker contained in the cyclopentylene bonded in the para position to the R group, and Z is a CN bonded to the cyclopentylene bonded in the para position; • Q1 is C, Q2 is C, Q3 is N, P is N, L1 is CH2, R is benzene, _L2 is a cyclopentylene or a methylene bond linker contained in the cyclopentylene bonded in the para position to the R group, and Z is a CN bonded to the cyclopentylene bonded in the 3rd position; • _Q1 is C, _Q2 is C, _Q3 is N, P is N, _L1 is CH2, R is benzene, L _Q2 is a cyclopentyl group bonded to the para position of the R group, and Z is CN bonded to the 3-position of the cyclopentyl group; and • _Q1 is C, _Q2 is C, _Q3 is N, P is N, _L1 is CH2, R is benzene, _L2 is a cyclopentyl group having a methylene bond linker in the para position at the 1-position bonded to the R group, and Z is CN bonded to the 3-position of the cyclopentyl group.

Additional embodiments include the following: • Q ₁ is C, Q ₂ is N, Q ₃ is C, P is N, L ₁ is CH 2, B is 1H-indene, and Z is CN; and • Q ₁ is C, Q ₂ is C, Q ₃ is N, P is N, L ₁ is CH 2, B is 2,3-dihydro-1H-indene, and Z is CN. Compounds of Formula IB

In additional embodiments (optionally in combination with any other embodiments described herein), the disclosure provides a compound of formula (IB) or a pharmaceutically acceptable salt thereof: (IB)

Q ₁ , Q ₂ , and Q ₃ are independently selected from the group consisting of CR ⁵ , N, O, and S.

^R5 is selected from the group consisting of H, _C1 - _C6 -alkyl, ^ORa , -( _C1 - _C6 -alkyl) ^ORa , and _C3 - _C10 -cycloalkyl, wherein each ^Ra is independently selected from the group consisting of H, _C1 - _C6 -alkyl, and _C1 - _C6 -haloalkyl.

P is C or N.

L ¹ is -SO ₂ - or -C ₁ -C ₈ -alkylene-.

is a divalent monocyclic or bicyclic moiety selected from the group consisting of C ₃ -C ₁₀ -cycloalkyl, C ₆ -C ₁₀ -aryl, 3 to 10 membered heterocycloalkyl (wherein 1 to 4 ring members are independently selected from N, O, and S), 5 to 10 membered heteroaryl (wherein 1 to 4 heteroaryl members are independently selected from N, O, and S), and fused combinations thereof.

^L2 is selected from the group consisting of a bond, _-C1 - _C8 -alkylene, _-C2 - _C8 -alkenylene, _-C2 - _C8 -alkynylene, and a divalent moiety selected from the group consisting of _C3 - _C10 -cycloalkyl, _C3 - _C10 -cycloalkenyl, _C6 - _C10 -aryl, 5- to 10-membered heteroaryl (wherein 1 to 4 heteroaryl members are independently selected from N, O, and S), ( _C1 - _C8 -alkyl) _C3 - _C10 -cycloalkyl, ( _C1 - _C8 -alkyl) _C3 - _C10 -cycloalkenyl, ( _C1 - _C8 -alkyl) _C6 - _C10 -aryl, and ( _C1 - _C8 -alkyl) 5-10 membered heteroaryl (wherein 1-4 heteroaryl members are independently selected from N, O, and S), wherein the cycloalkyl, cycloalkenyl, aryl, and heteroaryl are monocyclic or bicyclic.

L ² is optionally substituted with 1 to 3 substituents selected from the group consisting of C ₁ -C ₆ -alkyl, C ₁ -C ₆ -haloalkyl, and CN.

Z is selected from the group consisting of ^-ORc , -OC(O) ^Rc , -OC(O) ^{NRcRd ,} -S(O) _0-2Rc , ^-CN , -C(O) ^Rc , ^-C (O)ORc, -C(O) ^{NRcRd ,} -C(S) ^Rc _, -NRcRd ^, = ^NRc , ^{-NRcC (} O) ^NRcRd , ^-NRcCO2Rd , ^-NRc -NO, _-NO2 , -NRc ^{- ORd} , -N=C=O, -N= ^C =S ^, and ^{-NRc - NRcRd} .

Each example of R ^c and R ^d is independently selected from H, C ₁ -C ₆ -alkyl, C ₁ -C ₆ -haloalkyl, and C ₆ -C ₁₀ -aryl. Compounds of Formula IB-1

In additional embodiments (optionally in combination with any other embodiments described herein), the disclosure provides a compound of formula (IB-1) or a pharmaceutically acceptable salt thereof: (IB-1)

Q ₁ , Q ₂ , and Q ₃ are independently selected from the group consisting of CR ⁵ , N, O, and S.

^R5 is selected from the group consisting of H, _C1 - _C6 -alkyl, ^ORa , -( _C1 - _C6 -alkyl) ^ORa , and _C3 - _C10 -cycloalkyl, wherein each ^Ra is independently selected from the group consisting of H, _C1 - _C6 -alkyl, and _C1 - _C6 -haloalkyl.

P is C or N.

R ^e and R ^f are each independently selected from the group consisting of H, C ₁ -C ₆ -alkyl, and C ₁ -C ₆ -haloalkyl. Compounds of Formula IC

In additional embodiments (optionally in combination with any other embodiments described herein), the present disclosure provides a compound of formula (IC) or a pharmaceutically acceptable salt thereof: (IC)

^D1 is N or ^CR1 , ^D2 is N or ^CR2 , ^D3 is N or ^CR3 , and ^D4 is N or ^CR4 , wherein no more than three of ^D1 , ^D2 , ^D3 , and ^D4 are N at the same time.

R ¹ , R ² , R ³ , and R ⁴ are independently selected from the group consisting of H, C ₂ -C ₆ -alkenyl, C ₁ -C ₆ -halogenalkyl, halogen, NR ^a R ^b , OR ^a , -NR ^a C(O)R ^b , -C(O)R ^a , -C(O)halogen, -OC(O)R ^a , -OC(O)OR ^a , -C(O)OR ^a , C ₆ -C ₁₀ -aryl, CN, -S(O) _0-2 R ^a , -S(O) ₂ OR ^a , and NO ₂ .

Each of ^Ra and ^Rb is independently selected from the group consisting of H, _C1 - _C6 -alkyl, and _C1 - _C6 -haloalkyl.

R ^c1 , R ^c2 , and R ^c3 are independently selected from the group consisting of H, C ₁ -C ₆ -alkyl, and C ₁ -C ₆ -haloalkyl.

Q ₁ , Q ₂ , and Q ₃ are independently selected from the group consisting of CR ⁵ , N, O, and S.

R ⁵ is selected from the group consisting of H, C ₁ -C ₆ -alkyl, OR ^a , —(C ₁ -C ₆ -alkyl)OR ^a , and C ₃ -C ₁₀ -cycloalkyl.

P is C or N.

L ¹ is -SO ₂ - or -C ₁ -C ₈ -alkylene-.

is a divalent monocyclic or bicyclic moiety selected from the group consisting of C ₃ -C ₁₀ -cycloalkyl, C ₆ -C ₁₀ -aryl, 3 to 10 membered heterocycloalkyl (wherein 1 to 4 ring members are independently selected from N, O, and S), 5 to 10 membered heteroaryl (wherein 1 to 4 heteroaryl members are independently selected from N, O, and S), and fused combinations thereof.

^L2 is selected from the group consisting of a bond, _-C1 - _C8 -alkylene, _-C2 - _C8 -alkenylene, _-C2 - _C8 -alkynylene, and a divalent moiety selected from the group consisting of _C3 - _C10 -cycloalkyl, _C3 - _C10 -cycloalkenyl, _C6 - _C10 -aryl, 5- to 10-membered heteroaryl (wherein 1 to 4 heteroaryl members are independently selected from N, O, and S), ( _C1 - _C8 -alkyl) _C3 - _C10 -cycloalkyl, ( _C1 - _C8 -alkyl) _C3 - _C10 -cycloalkenyl, ( _C1 - _C8 -alkyl) _C6 - _C10 -aryl, and ( _C1 - _C8 -alkyl) 5-10 membered heteroaryl (wherein 1-4 heteroaryl members are independently selected from N, O, and S), wherein the cycloalkyl, cycloalkenyl, aryl, and heteroaryl are monocyclic or bicyclic.

L ² is optionally substituted with 1 to 3 substituents selected from the group consisting of C ₁ -C ₆ -alkyl, C ₁ -C ₆ -haloalkyl, and CN.

Z is selected from the group consisting of ^-ORc , -OC(O) ^Rc , -OC(O) ^{NRcRd ,} -S(O) _0-2Rc , ^-CN , -C(O) ^Rc , ^-C (O)ORc, -C(O) ^{NRcRd ,} -C(S) ^Rc _, -NRcRd ^, = ^NRc , ^{-NRcC (} O) ^NRcRd , ^-NRcCO2Rd , ^-NRc -NO, _-NO2 , -NRc ^{- ORd} , -N=C=O, -N= ^C =S ^, and ^{-NRc - NRcRd} .

Examples of R ^c and R ^d are independently selected from H, C ₁ -C ₆ -alkyl, C ₁ -C ₆ -haloalkyl, and C ₆ -C ₁₀ -aryl.

At least one of D ¹ , D ² , D ³ , or D ⁴ is not CH or at least one of R ^c1 , R ^c2 , or R ^c3 is not H.

Illustrative embodiments of the present disclosure pertain to the specific compounds presented in the Tables and Examples described herein.

The present disclosure also provides a pharmaceutical composition comprising a therapeutically effective amount of one or more compounds as described herein, or a pharmaceutically acceptable salt, stereoisomer, and/or tautomer thereof (admixed with a pharmaceutically acceptable carrier). In some embodiments, the composition further contains (according to recognized pharmaceutical formulation practices) one or more additional therapeutic agents, pharmaceutically acceptable excipients, diluents, adjuvants, stabilizers, emulsifiers, preservatives, colorants, buffers, flavor enhancers. The pharmaceutical composition can be administered by any suitable means, resulting in a concentration of the compound in the subject that is effective for treating a disease or condition suitable for treatment with the compounds of the present disclosure.

In some embodiments, the compound is present in an amount of 1 to 95% by weight of the total weight of the composition. The "therapeutically effective amount" of the compound or its pharmaceutically acceptable salt, stereoisomer, and/or tautomer is determined by such considerations and is the minimum amount necessary to induce PKal inhibition. This amount may be lower than the amount that is toxic to normal cells or the entire subject. Generally, the initial therapeutically effective amount of the compound of the present disclosure (or its pharmaceutically acceptable salt, stereoisomer, or tautomer) administered is in the range of about 0.001 to about 200 mg/kg or about 0.1 to about 20 mg/kg of patient weight per day, and the typical initial range is about 0.3 to about 15 mg/kg/day. Oral unit dosage forms (such as tablets and capsules) may contain about 0.1 mg to about 1000 mg of the disclosed compound (or its pharmaceutically acceptable salt, stereoisomer, or tautomer). In another embodiment, such dosage forms contain about 50 mg to about 500 mg of the disclosed compound (or its pharmaceutically acceptable salt, stereoisomer, or tautomer). In yet another embodiment, such dosage forms contain about 25 mg to about 200 mg of the disclosed compound (or its pharmaceutically acceptable salt, stereoisomer, or tautomer). In yet another embodiment, such dosage forms contain about 10 mg to about 100 mg of the disclosed compound (or its pharmaceutically acceptable salt, stereoisomer, or tautomer). In a further embodiment, such dosage forms contain about 5 mg to about 50 mg of a compound of the disclosure (or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof). In other embodiments, the compound is administered in a dosage form for administration into or around the eye, as described herein, for which a therapeutically effective amount of the compound may be in the range of about 0.0005 mg/kg to about 0.005 mg/kg, about 0.0007 mg/kg to about 0.004 mg/kg, or about 0.001 mg/kg to about 0.003 mg/kg of patient weight. In any of the foregoing embodiments, the dosage form may be administered once a day or twice a day.

Although the attending physician will ultimately select the appropriate dosage and dosing regimen, in additional embodiments, the therapeutically effective amount of the compounds described herein may be, for example, in the range of 0.0035 μg to 20 μg/kg body weight/day or 0.010 μg to 140 μg/kg body weight/week. In some embodiments, the therapeutically effective amount is in the range of 0.025 μg to 10 μg/kg, for example, at least 0.025, 0.035, 0.05, 0.075, 0.1, 0.25, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 5.0, 6.0, 7.0, 8.0, or 9.0 μg/kg body weight administered daily, every other day, or twice a week. In some embodiments, a therapeutically effective amount may be in the range of 0.05 μg to 20 μg/kg, for example at least 0.05, 0.7, 0.15, 0.2, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 10.0, 12.0, 14.0, 16.0, or 18.0 μg/kg of body weight administered daily, every other day, or twice a week. In some embodiments, a therapeutically effective amount of a compound may be in the range of 100 μg/m ² to 100,000 μg/m ² (of the subject's body surface area), for example administered daily, every other day, once a week, or every other week. In some embodiments, the therapeutically effective amount is in the range of 1000 μg/m ² to 20,000 μg/m ² , e.g., at least 1000, 1500, 4000, or 14,000 μg/m ² of the compound administered daily, every other day, twice weekly, weekly, or every other week.

In some embodiments, the compounds of the present disclosure are administered to adult humans at a daily dose of about 0.01 mg to 1000 mg (e.g., 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35 , 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mg).

In certain embodiments, the compounds described herein, or their pharmaceutically acceptable salts or solvents, are substantially pure, i.e., they contain less than about 5%, or less than about 2%, or less than about 1%, or less than about 0.5%, or less than about 0.1% of other small organic molecules, such as unreacted intermediates or synthesis by-products produced, for example, in one or more steps of the synthesis method.

In various embodiments, the compositions are provided in dosage forms suitable for oral, parenteral (e.g., intravenous, intramuscular, subcutaneous, intraarterial), buccal, sublingual, rectal, dermal, nasal, vaginal, intranasal, inhalation, transdermal, ocular, intraosseous, otic, or intracranial routes of administration. Therefore, in some embodiments, the dosage form of the composition is selected from tablets, capsules, tablets, powders, granules, suspensions, emulsions, solutions, gels (including hydrogels), pastes, patches, ointments, creams, plasters, drenches, osmotic delivery devices, suppositories, enemas, injections, implants, sprays, and aerosols. Pharmaceutical compositions are formulated in accordance with traditional pharmaceutical practice (see, e.g., Remington: The Science and Practice of Pharmacy, 20th edition, 2000, ed. A.R. Gennaro, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York).

The pharmaceutical composition can be formulated to release the active compound immediately after administration or to release the active compound at any predetermined time or any period after administration (e.g., a controlled release formulation). Examples of controlled release formulations include (i) formulations that produce a substantially constant concentration of the disclosed agent in the body over an extended period of time; (ii) formulations that produce a substantially constant concentration of the disclosed agent in the body over an extended period of time after a predetermined delay time; (iii) formulations that maintain the effect of the agent for a predetermined period of time (by maintaining a relatively constant level of the agent in the body) while minimizing fluctuations in the plasma level of the agent (such as a controlled release formulation). (iv) formulations that localize the effect of an agent, such as spatial placement of a controlled-release composition adjacent to or in a diseased tissue or organ; (v) formulations that achieve convenience of administration, such as administration of a composition once a week or every two weeks; and (vi) formulations that target the effect of an agent by using a carrier or chemical derivative to deliver the compound to a specific target cell type. In some embodiments, for compounds that have a narrow absorption window in the gastrointestinal tract or have a relatively short biological half-life, it is desirable to administer the compound in the form of a controlled-release formulation.

In some embodiments, controlled release is achieved by appropriate selection of various formulation parameters and ingredients, including, for example, various types of controlled release compositions and coatings. In some embodiments, the compound is formulated with an appropriate excipient into a pharmaceutical composition that releases the compound in a controlled manner after administration. Examples include single or multiple unit tablet or capsule compositions, oil solutions, suspensions, emulsions, microcapsules, molecular complexes, microspheres, nanoparticles, patches, and liposomes.

Pharmaceutical compositions comprising compounds as described herein can be administered parenterally by injection, infusion, or implantation (e.g., intraocularly, subcutaneously, intravenously, intramuscularly, intraperitoneally) via dosage forms, formulations, or by suitable delivery devices or implants containing known, non-toxic, pharmaceutically acceptable carriers and adjuvants. The formulations and preparations of such compositions are well known to those with ordinary knowledge in the art of pharmaceutical formulations.

Suitable oral compositions as described herein include, but are not limited to, tablets, lozenges, lachrymal lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups, or elixirs.

In another embodiment, also encompassed are pharmaceutical compositions suitable for single unit doses, which comprise a compound of the present disclosure or a pharmaceutically acceptable stereoisomer, salt, or tautomer thereof and a pharmaceutically acceptable carrier.

The compositions of the present disclosure suitable for oral use can be prepared according to any method known to the art of pharmaceutical composition manufacturing technology. For example, the liquid formulation of the compound of the present disclosure contains one or more agents selected from the group consisting of sweeteners, flavor enhancers, coloring agents, and preservatives to provide a pharmaceutically palatable preparation of the compound of the present disclosure.

For tablet compositions, the compounds of the present disclosure are blended with non-toxic pharmaceutically acceptable excipients for the manufacture of tablets. Examples of such excipients include, but are not limited to, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrants such as corn starch, or alginic acid; binders such as starch, gelatin, or gum arabic; and lubricants such as magnesium stearate, stearic acid, or talc. Tablets may be uncoated or they may be coated by known coating techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained therapeutic effect over a desired period of time. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed.

Formulations for oral use may also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent (e.g., calcium carbonate, calcium phosphate, or kaolin), or as soft gelatin capsules in which the active ingredient is mixed with water or an oil medium (e.g., peanut oil, liquid paraffin, or olive oil).

For aqueous suspensions, the compounds of the present disclosure are mixed with a suitable excipient for maintaining a stable suspension. Examples of such excipients include, but are not limited to, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and gum arabic.

The oral suspension may also contain a dispersant or a wetting agent such as a natural phosphatide (e.g., lecithin) or a condensation product of an alkylene oxide with a fatty acid (e.g., polyoxyethylene stearate) or a condensation product of an alkylene oxide with a long chain fatty alcohol (e.g., heptadecaethyleneoxycetanol), or a condensation product of an alkylene oxide with a partial ester derived from a fatty acid and a hexanol (e.g., polyoxyethylene sorbitan monooleate), or a condensation product of an alkylene oxide with a partial ester derived from a fatty acid and a hexanol (e.g., polyethylene sorbitan monooleate). The aqueous suspension may also contain one or more preservatives (for example ethyl p-hydroxybenzoate or n-propyl p-hydroxybenzoate), one or more coloring agents, one or more flavoring agents, and one or more sweetening agents (such as sucrose or saccharin).

Oily suspensions can be prepared by suspending the compounds of the present disclosure in vegetable oils (e.g., peanut oil, olive oil, sesame oil, or coconut oil), or in mineral oils (e.g., liquid wax). Oily suspensions may contain a thickening agent, such as beeswax, hard wax, or cetyl alcohol.

Sweeteners (such as those listed above) and flavor enhancers may be added to provide palatable oral preparations. These compositions may be preserved by the addition of an antioxidant (ascorbic acid).

Dispersible powders and granules suitable for preparing aqueous suspensions by adding water provide the compounds of the present disclosure in admixture with a dispersant or lubricant, a suspending agent, and one or more preservatives. Suitable dispersants or lubricants and suspending agents are exemplified by those mentioned above. Additional excipients (e.g., sweeteners, flavoring agents, and coloring agents) may also be present.

The pharmaceutical composition disclosed herein can also be in the form of an oil-in-water emulsion. The oily phase can be a vegetable oil (e.g., olive oil or peanut oil) or a mineral oil (e.g., liquid paraffin) or a mixture of these. Suitable emulsifiers can be natural gums (e.g., gum arabic or tragacanth), natural phospholipids (e.g., soybeans, lecithin), and esters or partial esters derived from fatty acids and hexitol, anhydrides (e.g., sorbitan monooleate), and condensation products of these specific esters and ethylene oxide (e.g., polyoxyethylene sorbitan monooleate). Emulsions can also contain sweeteners and flavor enhancers.

Syrups and syrups may be formulated with sweeteners (e.g., glycerol, propylene glycol, sorbitol, or sucrose). Such formulations may also contain demulcents, preservatives, and flavor enhancers and colorants. The pharmaceutical composition may be in the form of a sterile injection, an aqueous suspension, or an oily suspension. This suspension may be prepared according to known techniques using the appropriate dispersants or lubricants and suspending agents mentioned above. Sterile injectable preparations may also be sterile injectable solutions or suspensions in non-toxic parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol. Other acceptable vehicles and solvents that may be used include water, Ringer's solution, and isotonic sodium chloride solution. In addition, it is customary to employ sterile, non-volatile oils as solvents or suspending media. For this purpose, any non-irritating, non-volatile oil may be employed, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are useful in the preparation of injectables.

The compounds of the present disclosure may be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials are cocoa butter and polyethylene glycols.

Compositions for parenteral administration are administered in a sterile medium. Depending on the vehicle used and the concentration of the drug in the formulation, parenteral formulations may be suspensions or solutions containing dissolved drug. Adjuvants (such as local anesthetics, preservatives, and buffering agents) may also be added to parenteral compositions.

In some embodiments, the composition is specifically adapted for administration into or around the eye. For example, the composition can be adapted for use as eye drops, or injected into the eye, such as using peribulbar or intravitreal injection. Such compositions should be sterile and substantially free of endotoxins, and within an acceptable pH range. In some embodiments, a formulation without a preservative is used. Formulations for ocular administration are known in the art, see, for example, Ocular Therapeutics and Drug Delivery: A Multi-Disciplinary Approach, Reddy, Ed. (CRC Press 1995); Kaur and Kanwar, Drug Dev Ind Pharm. 2002 May; 28(5):473-93; Clinical Ocular Pharmacology, Bartlett et al. (Butterworth-Heinemann; 4th edition (Mar. 15, 2001)); and Ophthalmic Drug Delivery Systems (Drugs and the Pharmaceutical Sciences: a Series of Textbooks and Monographs), Mitra (Marcel Dekker; 2nd Rev&Ex edition (Mar. 1, 2003)).

Compositions for parenteral use may be provided in unit dosage form (e.g., in single-dose ampoules), or in vials containing several doses, to which a suitable preservative may be added (see below). The composition may be in the form of a solution, suspension, emulsion, infusion device, or implantable delivery device, or it may be presented as a dry powder for reconstitution with water or another suitable vehicle before use. In addition to the active agent(s), the composition may include a suitable parenterally acceptable carrier and/or formulation. The active agent(s) may be incorporated into microspheres, microcapsules, nanoparticles, liposomes, or the like for controlled release. Additionally, the composition may include a suspending agent, a solubilizing agent, a stabilizer, a pH adjuster, a osmotic pressure adjuster, and/or a dispersing agent.

In some embodiments, the pharmaceutical composition disclosed herein is in a form suitable for sterile injection. To prepare such compositions, the active agent(s) are dissolved or suspended in a parenteral acceptable liquid medium. Acceptable vehicles and solvents that can be used include water, water adjusted to a suitable pH by adding an appropriate amount of hydrochloric acid, sodium hydroxide, or a suitable buffer, 1,3-butylene glycol, Ringer's solution, dextrose solution, and isotonic sodium chloride solution. Aqueous formulations may also contain one or more preservatives (e.g., methyl parahydroxybenzoate, ethyl parahydroxybenzoate, or n-propyl parahydroxybenzoate). In cases where the solubility of the compound in water is limited, a solubility enhancer or solubilizing agent may be added, or the solvent may include 10 to 60% w/w propylene glycol.

The pharmaceutical composition can be administered to a subject in a single dose or in multiple doses. For example, the compounds described herein can be administered once a week, or 2, 3, 4, 5, 6, 7, 8, 10, 15, 20, or more weeks. It should be understood that for any particular subject, the specific dosage regimen should be adjusted over time according to the individual needs and the professional judgment of the health care provider administering or supervising the administration of the compound. For example, if a lower dose does not provide sufficient biological activity (e.g., in the treatment of a disease or condition described herein), the dose of the compound can be increased. Conversely, the dose of the compound can be reduced, for example, if the disease or condition is reduced or eliminated, or to reduce undesirable side effects. Use and Treatment Methods

As an advantage of the present disclosure, the compounds described herein are potent inhibitors of plasma microangiotensin, i.e., the compounds can reduce the activity of plasma microangiotensin. In various embodiments, the compounds described herein can be characterized by an inhibition constant _IC50 (half maximal inhibitory concentration) of no greater than 500 nM (e.g., less than 500, 450, 400, 350, 300, 250, 200, 150, 100, 50, 10, 1, or 0.1 nM).

In one embodiment, the present disclosure provides a method for inhibiting plasma microangiotensin. In some embodiments, the method comprises contacting PKal with a compound of the present disclosure in an amount effective to inhibit the activity of PKal. In some embodiments, the compounds of the present disclosure inhibit PKal activity with an IC50 value in the range of 0.1 to 500 nM (e.g., 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, ₄₅₀ , or 500 nM). It will be understood that all ranges in the present disclosure include one or both endpoints, all values between the endpoints (up to one significant digit), and any sub-ranges between the endpoints. In some embodiments, the compounds of the present disclosure inhibit PKal activity with an _IC50 of less than or equal to 500 nM, such as less than or equal to 500, 450, 400, 350, 300, 250, 200, 150, 100, 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1 nM.

In some embodiments, the method comprises administering to a subject an effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof. In some embodiments, the methods inhibit PKal activity in vivo with an _IC50 (half maximal inhibitory concentration) value in the range of 0.1 to 500 nM (e.g., 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, or 500 nM). In some embodiments, the methods inhibit PKal activity in vivo with an _IC50 of greater than 100 nM, such as 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 nM.

In additional embodiments, the present disclosure provides a method for treating a subject suffering from a disease or condition in a subject. The method comprises administering to the subject an effective amount of a compound as disclosed herein or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the compound is selected from the group consisting of any compound in Table 1, a combination thereof, and a pharmaceutically acceptable salt thereof.

In various embodiments, the disease or condition is selected from the group consisting of ischemic stroke, hemorrhagic stroke, hypertension, retinopathy, diabetic retinopathy, nephropathy, cerebral edema suppression, pulmonary hypertension, inflammation, acute myocardial infarction, deep venous embolism, complications from fibrinolytic therapy, stroke, angina, vascular edema, sepsis, arthritis, complications of cardiopulmonary bypass, microvascular leak syndrome, inflammatory bowel disease, diabetes Vascular complications of disease, diabetic macular edema, macular degeneration, neuropathy, age-related macular degeneration, retinal vein occlusion, cerebral edema, ischemia-reperfusion injury, angiogenesis, asthma, acute allergy, Alzheimer's disease, Parkinson's disease, multiple sclerosis, glioblastoma multiforme, complications of fibrinolytic therapy, increased albumin secretion, macroalbuminuria, pain, muscle Atrophic lateral sclerosis, Kugelman's disease, epilepsy, brain trauma, high altitude cerebral edema, cancer, generalized intravascular coagulation, pancreatitis, inflammation, shock, hereditary vascular edema (HAE), uveitis, polyangiitis, acute respiratory distress syndrome (ARDS), thrombosis, vasculitis, Crohn's disease, ulcerative colitis, colitis, arteritis, glomerulonephritis, eczema, endometriosis, preeclampsia, malaria, arthritis, intermittent fever and recurrence Fever, Choger's disease, Raynaud's disease, systemic sclerosis, granulomatosis with polyangiitis, small vessel vasculitis, medium vessel vasculitis, large vessel vasculitis, pan vasculitis, systemic autoinflammatory disease, renal failure, cerebral malaria, Clarkson's disease (systemic vascular leak syndrome), Hantavirus infection, Hantavirus renal syndrome, Hantavirus pulmonary syndrome, virus-related inflammatory diseases, retinal vasculitis, uveitis, Eales' disease, Behcet's disease, sarcoidosis, whooping cough, coronavirus infection, and non-infectious posterior uveitis.

In some embodiments, the subject is an animal, such as a human or a non -human animal (e.g., a mammal), and is used interchangeably with "patient" when the subject is being treated medically by a health care provider.

The present disclosure also provides a combination pharmaceutical composition comprising: (a) at least one compound as disclosed herein or a pharmaceutically acceptable salt thereof, and (b) at least one inhibitor of inflammation, pain, or edema. In some embodiments, a pharmaceutical composition for oral delivery is provided, which comprises the combination pharmaceutical composition of the present disclosure. The present disclosure further provides tablets, capsules, oral delivery particles, injectable suspensions, and solutions comprising the combination pharmaceutical composition, as well as compositions for transpulmonary or transnasal delivery. Kits

In another embodiment , the present disclosure provides a kit comprising a compound as described herein or a pharmaceutically acceptable salt thereof. The kit also comprises instructions for a health care provider to administer the compound to a patient.

The following examples provide further embodiments of the present disclosure. These examples are illustrative and non-limiting. Although any compositions, compounds, and methods substantially similar to those described herein can be used in the practice or testing of the present disclosure, the compositions, compounds, and methods described are merely exemplary. Synthesis of Compounds

As described in the examples below, in certain illustrative embodiments, compounds are prepared according to the following general procedures. It will be recognized that although the general methods describe the synthesis of certain compounds of the present invention, the following general methods, and other methods known to those of ordinary skill in the art, can be applied to all compounds and subclasses and species of each of these compounds, as described herein. Additional compounds of the present invention are prepared by methods substantially similar to those described in the examples herein and known to those of ordinary skill in the art. Preparation of Intermediate Compounds Example 1A : Preparation of ethyl 1-(4-(2- cyanopropan-2- yl) benzyl)-1H- pyrazole-4- carboxylate (7_Int-5) Step-1. Synthesis of 2- methyl-2-( p-tolyl) propionitrile (7-Int-2) .

To a solution of 2-(p-tolyl)acetonitrile (200 g, 1524 mmol, 1.0 eq) in N-methyl-2-pyrrolidone (1000 mL, 5V) and tetrahydrofuran (1000 mL, 5V) was added sodium tert-butoxide (586.07 g, 6098 mmol, 4.0 eq) in portions at 0°C to 5°C in a nitrogen atmosphere. The reaction mixture (RM) was stirred at 0°C to 5°C for 30 min. Methyl iodide (865.62 g, 6098 mmol, 4.0 eq) was added dropwise to the RM at 0°C to 5°C. The RM was stirred at 0°C to RT for 2.5 h. After the reaction was completed, the RM was transferred to deionized water (400 mL) and the product was extracted with ethyl acetate (2000 mL). The combined organic fractions were washed with cold water (4x500 mL) to remove N-methyl-2-pyrrolidone, concentrated and purified by column chromatography (2-5% EtOAc in hexanes) to give compound 7-Int-2. ¹ H NMR (400 MHz, DMSO-d6) δ 1.651 (s, 6H), 2.298 (s, 3H), 7.226 d, J=8 Hz, 2H), 7.392 (d, J=6.8 Hz, 2H). Step-2. Synthesis of 2-(4-( bromomethyl) phenyl)-2- methylpropionitrile (7-Int-3) .

In a 10000 mL 4N round bottom flask (RBF) attached to a mechanical stirrer and condenser was poured 2-methyl-2-(p-tolyl)propionitrile (170 g, 1067 mmol, 1.0 eq) and carbon tetrachloride (3400 mL, 20V) at RT. AIBN (17.53 g, 106 mmol, 0.1 eq) was added to the RM. N-bromosuccinimide (209.04 g, 1174 mmol, 1.1 eq) was added to the RM in portions at RT. The resulting RM was heated to 90 °C and stirred for 2 h. Note: After the reaction was completed, the reaction mixture was cooled to RT. The RM was quenched into DM water (3400 mL) and the product was extracted with DCM (2x2000 mL). The combined organic fractions were concentrated and then purified by column chromatography (7% EtOAc in hexanes) to give compound (7_Int-3). ¹ H NMR (400 MHz, DMSO-d6) δ 1.654 (s, 6H), 4.714 (s, 2H), 7.507 (s, 4H). Step-3. Synthesis of ethyl 1-(4-(2- cyanopropan-2- yl) benzyl)-1H- pyrazole-4- carboxylate (7_Int-5) .

In a 3000 mL 4N RBF attached to a mechanical stirrer and condenser, 1H-pyrazole-4-carboxylic acid ethyl ester (70 g, 499 mmol, 1.0 eq), 2-(4-(bromomethyl)phenyl)-2-methylpropionitrile (130.84 g, 549 mmol, 1.1 eq), and acetone (700 mL, 10 V) were poured at RT. Cs ₂ CO ₃ (390.8 g, 1198 mmol, 2.4 eq) was added to the RM. The RM was heated to 60-65 °C and stirred for 6 h. After the reaction was complete, the reaction mixture was cooled to RT. The RM was filtered to remove Cs ₂ CO ₃ ; washed with EtOAc. The filtrate was concentrated and purified by column chromatography (15% ethyl acetate in hexanes) to give compound (7-Int-5). ¹ H NMR (400 MHz, DMSO-d6) δ 1.656 (s, 6H), 4.204 (q, J=6.8 Hz, 2H), 5.372 (s, 2H), 7.328 (d, J=8 Hz, 2H), 7.498 (d, J=8 Hz, 2H), 8.053 (s, 1H), 8.482 (s, 1H). Example 2A : Preparation of ethyl 1-(4-( cyanomethyl) benzyl)-1H- pyrazole-4- carboxylate (1-Int-4) Step-1. Synthesis of ethyl 1-(4- methylbenzyl)-1H- pyrazole-4- carboxylate (1_Int-2) .

In a 10000 mL 4N RBF attached to a mechanical stirrer and a condenser, 1H-pyrazole-4-carboxylic acid ethyl ester (250 g, 1783 mmol, 1.0 eq), 1-(bromomethyl)-4-methylbenzene (363.1 g, 1962 mmol, 1.1 eq), and acetone (6250 mL, 25 V) were poured at RT. Cs ₂ CO ₃ (390.8 g, 1198 mmol, 2.4 eq) was added to the RM. The RM was heated to 60-65 °C and stirred for 16 h. After the reaction was completed, the reaction mixture was cooled to RT. The RM was filtered to remove Cs ₂ CO ₃ ; washed with EtOAc; the filtrate was concentrated under reduced pressure to obtain a crude product. The crude was purified by trituration in hexanes to give compound (1_Int-2). ¹ H NMR (400 MHz, DMSO-d6) δ 1.227 (t, J=6.8 Hz, 3H), 2.247 (s, 3H), 4.176 (q, J=6.8 Hz, 2H), 5.282 (s, 2H), 7.144-7.136 (m, 4H), 7.828 (s, 1H), 8.390 (s, 1H). Step-2. Synthesis of ethyl 1-(4-( bromomethyl) benzyl)-1H- pyrazole-4- carboxylate (1_Int-3) .

In a 10000 mL 4N RBF attached to a mechanical stirrer and condenser, ethyl 1-(4-methylbenzyl)-1H-pyrazole-4-carboxylate (300 g, 1228 mmol, 1.0 eq) and 1,2-dichloroethane (6000 mL, 20 V) were poured at RT. Benzoyl peroxide (29.71 g, 122.8 mmol, 0.1 eq) was added to the RM. N-bromosuccinimide (240.4 g, 1350 mmol, 1.1 eq) was added to the RM in portions at RT. The resulting RM was heated to 90 °C and stirred for 4 h. After the reaction was complete, the reaction mixture was cooled to RT. The RM was diluted with DCM (2000 mL). The RM was washed with (2 x 4000 ml) saturated _Na2SO4 solution and concentrated under reduced pressure to obtain _the crude. The crude was purified by column chromatography (17 to 25% EtOAc in hexanes) to give compound (1-Int-3). ¹ H NMR (400 MHz, DMSO-d6) δ 1.270-1.235 (m, 3H), 4.207 (q, J=6.8 Hz, 2H), 4.682 (s, 2H), 5.364 (s, 2H), 7.243 (d, J=8.4 Hz, 4H), 7.421 (d, J=8.4 Hz, 2H), 7.868 (s, 1H), 8.478 (s, 1H). Step-3. Synthesis of ethyl 1-(4-( cyanomethyl) benzyl)-1H- pyrazole-4- carboxylate (1-Int-4) .

In a 5000 mL 4N RBF attached to a mechanical stirrer and condenser, ethyl 1-(4-(bromomethyl)benzyl)-1H-pyrazole-4-carboxylate (242 g, 499748 mmol, 1.0 eq), ACN (2420 mL, 10V), and Cs ₂ CO ₃ (488 g, 1497 mmol, 2.0 eq) were poured at RT followed by slow addition of TMSCN (334 g, 3369 mmol, 4.5 eq) at RT. The RM was heated to 80-85 °C and stirred for 16 h. After the reaction was complete, the reaction mixture was cooled to RT. The RM was filtered to remove Cs ₂ CO ₃ ; washed with EtOAc. The solid residue was discarded and the filtrate was concentrated under reduced pressure to give a crude product. The crude product was purified by column chromatography (12 to 15% EtOAc in hexanes) to give compound (1-Int-4). ¹ H NMR (400 MHz, DMSO-d6) δ 1.282-1.234 (m, 3H), 4.016 (s, 2H), 4.20 (q, J=7.2 Hz, 2H), 5.363 (s, 2H), 7.339-7.280 (m, 4H), 8.051 (s, 1H), 8.468 (s, 1H). Example 3A : Preparation of ethyl 1-(4-((3- cyano -1H- pyrrol -1- yl ) methyl ) benzyl )-1H- pyrazole -4- carboxylate (6-Int-1) Step -1. Synthesis of ethyl 1-(4-((3- cyano -1H- pyrrol -1- yl ) methyl ) benzyl )-1H- pyrazole -4- carboxylate (6-Int-1) .

A stirring solution of 1H-pyrrole-3-carbonitrile (0.2 g, 2.16 mmol, 1.0 eq) in DMF (2 mL, 10 V) was prepared and NaH (60% in mineral oil) (0.156 g, 3.24 mmol, 1.5 eq) was added at 0 °C. The RM was stirred at 0 °C for 30 min. 1-(4-(bromomethyl)benzyl)-1H-pyrazole-4-carboxylic acid ethyl ester (1_Int-3) (0.772 g, 2.39 mmol, 1.1 eq) was added to the RM at 0 °C. The RM was warmed to RT and stirred for 3 h. After the reaction was complete, the RM was quenched into water and extracted with ethyl acetate. The combined organic fractions were dried _{over Na2SO4} , concentrated and then purified by column chromatography (20% ethyl acetate in hexanes) to give compound (6-Int-1). MS (ES): 335.30 m/z [M+H]+, LCMS purity: 95.09%, ¹ H NMR (400 MHz, DMSO-d6) δ 1.246 (t, J=8 Hz, 3H), 4.194 (q, J=7.2 Hz, 2H), 5.336 (s, 2H), 6.987 (t, J=2.4 Hz, 1H), 7.267-7.208 (m, 4H), 7.699 (t, J=2 Hz, 1H), 7.845 (s, 1H), 8.448 (s, 1H). Example 4A : Preparation of 4-( aminomethyl)-2- fluorobenzamidine dihydrochloride (43-Int-5) Step-1. Synthesis of tributyl (4- cyano-2- fluorobenzyl) carbamate (43_Int-2) .

Preparation of 4-(aminomethyl)-2-fluorobenzonitrile (43_Int-1) (3 g, 19.97 mmol, 1.0 eq) in 1,4-dihydro- A stirred solution of 2-nitropropane (36 mL, 12V) and 2N NaOH (18 mL, 6V) was added and Boc anhydride (4.79 g, 21.97 mmol, 1.1 eq) was added at 0°C. The reaction mixture was stirred at RT for 3 h. After the reaction was completed, the RM was quenched in water and extracted with ethyl _acetate . The combined organic fractions were dried over _Na2SO4 ; concentrated to give compound (43_Int-2). MS (ES): 251.00.m/z [M+H]+, LCMS purity: 100%, ¹ H NMR (400 MHz, DMSO-d6) δ 1.393 (s, 9H), 4.216 (d, J=5.2 Hz, 2H), 7.281 (t, J=7.5 Hz, 2H), 7.551 (s, 1H), 7.885 (t, J=8 Hz, 1H) Step-2. Synthesis of tributyl (3- fluoro-4-(N- hydroxycarbamimidoyl) benzyl) carbamate (43_Int-3) .

A stirred solution of (4-cyano-3-fluorobenzyl)carbamic acid tributyl ester (43_Int-2) (3.4 g, 13.58 mmol, 1.0 eq) and methanol (34 mL, 10V) was prepared and hydroxylamine hydrochloride (1.604 g, 23.09 mmol, 1.7 eq) and DIPEA (3.95 g, 23.09 mmol, 1.7 eq) were added at RT. The RM was stirred at 70 °C for 16 h. After completion of the reaction, the RM was evaporated and the residue was quenched in water and extracted with 10% _methanol in DCM. The combined organic fractions were dried over _Na2SO4 ; concentrated to give compound (43_Int-3). MS (ES): 284.10 m/z [M+H]+ LCMS purity: 81.45%, ¹ H NMR (400 MHz, DMSO-d6) δ 1.393 (s, 9H), 4.133 (d, J=5.2 Hz, 2H), 7.101-70.31 (m, 2H), 7.125 (s, 1H), 7.483-7.417 (m, 2H), 7.611 (t, J=8 Hz, 1H), 9.593 (s, 1H). Step-3. Synthesis of tributyl (4- carboxamidino-3- fluorobenzyl) carbamate (43_Int-4) .

A stirred solution of tributyl (3-fluoro-4-(N-hydroxycarbamimidoyl)benzyl)carbamate (43_Int-3) (3.5 g, 12.35 mmol, 1.0 eq) and methanol (35 mL, 10V) was prepared and ammonium formate (2.34 g, 37.06 mmol, 3.0 eq) and 10% Pd/C (0.175 g, 0.065% w/w) were added at RT. The RM was stirred at 70 °C under 20 kg H2 pressure in an autoclave for 16 h. After completion of the reaction, the RM was filtered through a celite bed and the filtrate was concentrated to give compound (43_Int-4). MS (ES): 267.80m/z [M+H]+ LCMS purity: 96.46%, ¹ H NMR (400 MHz, DMSO-d6) δ 1.394 (s, 9H), 4.195 (d, J=5.6 Hz, 2H), 7.244 (d, J=8 Hz, 2H), 7.254-7.233 (m, 3H), 7.639-7.580 (m, 3H), 8.415 (s, 1H). Step-4. Synthesis of 4-( aminomethyl)-2- fluorobenzamidine dihydrochloride (43_Int-5) .

A stirred solution of (4-carbamimidoyl-3-fluorobenzyl)carbamic acid tributyl ester (43_Int-4) (4.8 g, 16.83 mmol, 1.0 eq) and water (52 mL, 11 V) was prepared and concentrated HCl (16 mL, 3.3 V) was added at RT. The RM was stirred at RT for 3 h. After completion of the reaction, the RM was concentrated and triturated with methanol to give compound (43_Int-5). MS (ES): 168.13 m/z [M+H]+, ¹ H NMR (400 MHz, DMSO-d6) δ 4.141 (d, J=4 Hz, 2H), 7.658-7.155 (m, 3H), 7.737-7.686 (m, 2H), 8.772 (s, 1H), 9.564-9.440 (m, 2H). Example 5A : Preparation of 4-( aminomethyl)-3- fluorobenzamidine dihydrochloride (45_Int-5) Step-1. Synthesis of tributyl (4- cyano-2- fluorobenzyl) carbamate (45_Int-2) .

Preparation of 4-(aminomethyl)-3-fluorobenzonitrile (45_Int-1) (1 g, 6.65 mmol, 1.0 eq) in 1,4-dihydro- To a stirred solution of 2-nitro-1-oxane (12 mL, 12V) and 2N NaOH (6 mL, 6V) was added Boc anhydride (1.59 g, 7.32 mmol, 1.1 eq) at 0°C. The reaction mixture was stirred at RT for 3 h. After completion of the reaction, the RM was quenched in water and extracted with ethyl _acetate . The combined organic fractions were dried over _Na2SO4 ; concentrated to give compound (45_Int-2). ^1H NMR (400 MHz, DMSO-d6) δ 1.146 (s, 9H), 4.220 (d, J=5.2 Hz, 2H), 7.442 (t, J=7.5 Hz, 2H), 7.525 (s, 1H), 7.812 (t, J=8 Hz, 2H). Step-2. Synthesis of tributyl (2- fluoro-4-(N- hydroxycarbamimidoyl) benzyl) carbamate (45_Int-3) .

A stirred solution of (4-cyano-2-fluorobenzyl)carbamic acid tributyl ester (45_Int-2) (5.3 g, 21.17 mmol, 1.0 eq) and methanol (53 mL, 10V) was prepared and hydroxylamine hydrochloride (2.5 g, 36.0 mmol, 1.7 eq) and DIPEA (4.64 g, 36.0 mmol, 1.7 eq) were added at RT. The RM was stirred at 70 °C for 16 h. After completion of the reaction, the RM was evaporated and the residue was quenched in water and extracted with 10% _methanol in DCM. The combined organic fractions were dried over _Na2SO4 ; concentrated to give compound (45_Int-3). MS (ES): 284.21 m/z [M+H]+ Step-3. Synthesis of tributyl (4- carbamimidoyl-2- fluorobenzyl) carbamate (45_Int-4).

A stirred solution of tributyl (2-fluoro-4-(N-hydroxycarbamimidoyl)benzyl)carbamate (45_Int-3) (2.4 g, 8.47 mmol, 1.0 eq) and methanol (24 mL, 10 V) was prepared and ammonium formate (1.6 g, 25.41 mmol, 3.0 eq) and 10% Pd/C (0.16 g, 0.065% w/w) were added at RT. The RM was stirred at 70 °C under 20 kg H2 pressure in an autoclave for 16 h. After completion of the reaction, the RM was filtered through a celite bed and the filtrate was concentrated to give compound (45_Int-4). MS (ES): 268.18 m/z [M+H]+ Step-4. Synthesis of 4-( aminomethyl)-3- fluorobenzamidine dihydrochloride (45_Int-5) .

A stirred solution of (4-carbamimidoyl-2-fluorobenzyl)carbamic acid tributyl ester (45_Int-4) (2.0 g, 7.48 mmol, 1.0 eq) and water (22 mL, 11 V) was prepared and concentrated HCl (6.6 ml, 3.3 V) was added at RT. The RM was stirred at RT for 3 h. After completion of the reaction, the RM was concentrated and triturated with methanol to give compound (45_Int-5). MS (ES): 168.13 m/z [M+H]+, ¹ H NMR (400 MHz, DMSO-d6) δ 4.145 (s, 2H), 7.210 (s, 1H), 7.336 (s, 1H), 7.463 (s, 1H), 7.878-7.765 (m, 2H), 8.816 (s, 2H), 9.442 (s, 1H), 9.634 (s, 1H). Example 6A : Preparation of ethyl 1-(4-(3- cyanobenzyl ) benzyl )-1H- pyrazole -4- carboxylate (28-Int-4) Step-1. Synthesis of ethyl 1-(4- chlorobenzyl)-1H- pyrazole-4- carboxylate (28_Int-2)

A stirred solution of ethyl 1H-pyrazole-4-carboxylate (1 g, 7.14 mmol, 1 eq) and 1-(bromomethyl)-4-chlorobenzene (28_int-1) (1.5 g, 7.85 mmol, 1.1 eq) in acetone (10 mL, 10 V) was prepared and Cs ₂ CO ₃ (5.53 g, 17.14 mmol, 2.4 eq) was added at RT. The RM was heated to 65 °C and stirred for 4 h. After completion of the reaction, the RM was cooled at RT, quenched in water and extracted with ethyl acetate. The combined organic fractions were dried over Na ₂ SO ₄ ; concentrated and purified by flash column chromatography (50% ethyl acetate in hexanes) to give compound (28_Int-2). MS (ES): 265.30 m/z [M+], 266.30 [M+2]+, LCMS purity: 63%, ¹ H NMR (400 MHz, DMSO-d6) δ 400 MHz, DMSO-d6: δ 1.26 (t, J = 6.8 Hz, 3H), 4.28-4.210 (m, 2H), 5.38 (s, 2H), 7.30 (d, J = 8.40 Hz, 2H), 7.43 (d, J = 8.40 Hz, 2H), 7.88 (s, 1H), 8.49 (s, 1H). Step-2. Synthesis of 1-(4-(4,4,5,5- tetramethyl-1,3,2- diboron - 2 - yl ) benzyl)-1H- pyrazole-4- carboxylic acid ethyl ester (28_Int-3)

Preparation of ethyl 1-(4-chlorobenzyl)-1H-pyrazole-4-carboxylate (28_Int-2) (1.0 g, 3.78 mmol, 1.0 eq) in 1,4-dihydro-1H-pyrazole-4-carboxylate To a stirred solution in 2-(4-(2-pyridyl)-2-nitropropene (5 mL, 5V) was added bis(pinacol)diboron (1.14 g, 4.54 mmol, 1.2eq), KOAc (1.11 g, 11.36 mmol, 3eq), XPhosPdG2 (0.350 g, 0.37 mmol, 0.1eq), and water (0.5 mL, 0.5V) at RT. The RM was heated to 100 °C and stirred for 16 h. After completion of the reaction, the RM was cooled to RT, quenched in water and extracted with ethyl _acetate . The combined organic fractions were dried over _Na2SO4 ; concentrated and then purified by flash column chromatography (20% ethyl acetate in hexanes) to give compound (28_Int-3). MS (ES): 357.51 m/z [M+1]+, LCMS purity: 70%, ¹ H NMR (400 MHz, DMSO-d6) δ 1.26 (s, 12H), 1.26 (t, 3H), 4.228-4.210 (m, 2H), 5.38 (d, J = 12.80 Hz, 2H), 7.25 (d, J = 8.00 Hz, 2H), 7.65 (d, J = 8.00 Hz, 1H), 7.87 (s, 1H), 0.00 (s, 1H), 8.47 (s, 1H). Step -3. Synthesis of ethyl 1-(4-(3- cyanobenzyl ) benzyl )-1H- pyrazole -4- carboxylate (28_Int-4)

Preparation of ethyl 1-(4-(4,4,5,5-tetramethyl-1,3,2-diboron-2-yl)benzyl)-1H-pyrazole-4-carboxylate (28_Int-3) (0.300 g, 0.84 mmol, 1.0 eq) in 1,4-dihydro-1-(4-(4,4,5,5-tetramethyl-1,3,2-diboron-2-yl)benzyl)-1H-pyrazole-4-carboxylate To a stirred solution in 2-(bromomethyl)-1-nitropropane (3 mL, 10 V) was added 3-(bromomethyl)benzonitrile (0.181 g, 0.92 mmol, 1.1 eq), K3PO4 (0.535 g, 3.51 mmol, 3 eq), Pd(dppf)Cl2, DCM (0.068 g, 0.11 mol, 0.1 eq), and water (1.5 mL, 5 V) at RT. The RM was heated to 100 °C and stirred for 6 h. After the reaction was complete, the RM was cooled to RT, quenched in water and extracted with _ethyl acetate. The combined organic fractions were dried over _Na2SO4 ; concentrated and then purified by flash column chromatography (30% ethyl acetate in hexane) to give compound (28_Int-4). MS (ES): 346.39 m/z [M+1]+, LCMS purity: 100%, ¹ H NMR (400 MHz, DMSO-d6) δ 1.22 (s, 3H), 3.98 (s, 2H), 4.20 (q, J = 6.8 Hz, 2H), 5.31 (s, 2H), 7.254-7.193 (m, 4H), 7.49 (t, J = 8.00 Hz, 1H), 7.56 (d, J = 8.00 Hz, 1H), 0.00 (d, J = 7.60 Hz, 1H), 7.72 (s, 1H), 7.84 (s, 1H), 8.434 (s, 1H). Example 7A : Preparation of 4-( aminomethyl)-2,6 -difluorobenzamidine dihydrochloride (44_Int-8) Step-1. Synthesis of 2,6 -difluoro-4-( hydroxymethyl) benzonitrile (44_Int-2)

A stirred solution of 2,6-difluoro-4-formylbenzonitrile (44_Int-1) (8 g, 0.047 mmol, 1.0eq) in methanol (160 mL, 20V) was prepared and _NaBH4 (2.21 g, 0.047 mmol, 1eq) was added at 0 °C. The RM was stirred at RT for 1 h. After completion of the reaction, the RM was quenched in water and extracted with _DCM . The combined organic fractions were dried over _Na2SO4 ; concentrated to give compound (44_Int-2) . 1H NMR (400 MHz, DMSO-d6) δ 4.60 (d, J= 8 Hz, 2H), 5.72 (t, J=9.1 Hz, 1H), 7.34 (d, J= 8 Hz, 2H). Step-2. Synthesis of 2,6- difluoro-4-( hydroxymethyl) benzonitrile (44_Int-3)

A stirred solution of 2,6-difluoro-4-(hydroxymethyl)benzonitrile (44_Int-2) (7.5 g, 0.0443 mmol, 1.0eq) in MTBE (75 mL, 10V) was prepared and PBr ₃ (14.37 g, 0.0532 mmol, 1.2eq) was added at 0°C. The RM was stirred at RT for 2h. After completion of the reaction, the RM was quenched in cold water and extracted with ethyl acetate. The combined organic fractions were dried over Na ₂ SO ₄ ; concentrated to give compound (44_Int-3) . 1H NMR (400 MHz, DMSO-d6) δ 4.721 (s, 2H), 7.564 (d, J=8.8 Hz, 2H). Step-3. Synthesis of 4-( aminomethyl)-2,6- difluorobenzonitrile (44_Int-4)

Ammonia gas was purged in methanol at 0 °C for 30 min. A solution of 2,6-difluoro-4-(hydroxymethyl)benzonitrile (44_Int-3) (9.0 g, 0.06 mmol, 1.0eq) in MeOH (40 mL, 10V) was added to RM and stirred at 0 °C for 4 h. After completion of the reaction, RM was concentrated to give compound (44_Int-4) . MS (ES): 169 m/z [M+1]+, LCMS purity: 84%. Step-4. Synthesis of tributyl (4- cyano-3,5- difluorobenzyl) carbamate (44_Int-5)

Preparation of 4-(aminomethyl)-2,6-difluorobenzonitrile (44_Int-4) (10 g, 59.5238 mmol, 1.0 eq) in 1,4- The stirred solution was added to 2N NaOH (60 mL, 6V) and Boc-anhydride (14.27 gm, 0.065 mmol, 1.1 eq) at RT and stirred for 5 h. After the reaction was completed, the RM was quenched in cold water and extracted with ethyl acetate. The combined organic fractions were dried over Na ₂ SO ₄ ; concentrated to give compound (44_Int-5) 1H NMR (400 MHz, DMSO-d6) δ 1.39 (s, 9H), 4.52 (s, 2H), 7.27 (d, J =8.8 Hz, 2H). Step-5. Synthesis of tributyl (3,5 -difluoro-4-(N- hydroxycarbamimidoyl) benzyl) carbamate (44_Int-6)

A stirred solution of (4-cyano-3,5-difluorobenzyl)carbamic acid tributyl ester (44_Int-5) (6 g, 22.388 mmol, 1.0eq) in MeOH (60 mL, 10V) was prepared. Hydroxylamine hydrochloride (2.6 gm, 38.0596 mmol, 1.7eq) and DIPEA (6.5 mL, 38.0596 mmol, 1.7eq) were added at RT and stirred at 70 °C for 16 h. After completion of the reaction, the RM was cooled to RT. The RM was then quenched in water and extracted with _DCM . The combined organic fractions were dried over _Na2SO4 ; concentrated to give compound (44_Int-6) . MS (ES): 302 m/z [M+1]+, LCMS purity: 65%. Step-6. Synthesis of tributyl (4- carbamimidoyl-3,5 -difluorobenzyl) carbamate (44_Int-7)

A stirred solution of tributyl (3,5-difluoro-4-(N-hydroxycarbamimidoyl)benzyl)carbamate (44_Int-6) (6 g, 19.933 mmol, 1.0eq) in MeOH (60 mL, 10V) was prepared. Ammonium chloride (5.38 g, 99.66 mmol, 5.0eq) and iron (5.40 g, 99.66 mmol, 5.0eq) were added at RT. The RM was cooled to 0 °C and acetic acid (30 mL, 5V) was added dropwise. The RM was then heated to 70 °C and stirred for 16 h. After the reaction was complete, the RM was cooled to RT. The RM was concentrated and quenched in cold water and then slowly alkalized to pH ~10. The solid was isolated by filtration and extracted with DCM. The combined organic fractions were dried over Na ₂ SO ₄ and concentrated to give compound (44_Int-7) . MS (ES): 286 m/z [M+1]+, LCMS purity: 65%. Step-7. Synthesis of 4-( aminomethyl)-2,6 -difluorobenzamidine (44_Int-8)

A stirred solution of (4-carbamimidoyl-3,5-difluorobenzyl)carbamic acid tributyl ester (44_Int-7) (2.2 g, 25.473 mmol, 1.0 eq) in water (24 mL, 11 V) was prepared and concentrated HCl (7.26 mL, 3.3 V) was added at RT and stirred for 3 h. After completion of the reaction, the RM was concentrated and triturated in methanol to give compound (44_Int-8) . MS (ES): 231 m/z [M+1]+, LCMS purity: 67%. Example 9A : Preparation of methyl 1-(4-(2- cyanopropan-2- yl) benzyl)-5-( methoxymethyl)-1H- pyrazole-4- carboxylate (82_Int-3) Step-1. Synthesis of methyl 5-( methoxymethyl)-1H- pyrazole-4- carboxylate (82_Int-2)

A solution of methyl 4-methoxy-3-oxobutyrate (82_Int-1) (15 g, 102 mmol, 1.0 eq) and dimethylformamide dimethylacetal (12.23 g, 102 mmol, 1.0 eq) was prepared at 110 °C and stirred for 1 h. The RM was cooled to RT followed by the addition of ethanol (150 mL, 10 V) and hydrazine hydrate (99%) (5.13 g, 102 mmol, 1.0 eq). The RM was heated to 70 °C and stirred for 2 h. After completion of the reaction, the RM was evaporated and then purified by column chromatography (25-30% ethyl acetate in hexanes) to give compound (82_Int-2) . 1H NMR (400 MHz, DMSO-d6) δ 3.258 (s, 3H), 3.758 (s, 3H), 4.702 (s, 2H), 7.837 (s, 1H), 8.281 (s, 1H). Step -2. Synthesis of 1-(4-(2- cyanopropan -2- yl ) benzyl )-5-( methoxymethyl )-1H- pyrazole -4- carboxylic acid methyl ester (82_Int-3)

A stirred solution of methyl 5-(methoxymethyl)-1H-pyrazole-4-carboxylate (82_Int-2) (1.0 g, 5.87 mmol, 1.0 eq) in DMF (10 mL, 10 V) was prepared and NaHMDS (1M in THF) (5.8 mL, 5.87 mmol, 1.0 eq) was added at 0 °C and stirred for 30 min. Then 2-(4-(bromomethyl)phenyl)-2-methylpropionitrile (7_Int-3) (1.39 g, 5.87 mmol, 1.0 eq) was added to the RM at 0 °C and stirred for additional 16 h. After the reaction was complete, the RM was quenched into water and extracted with ethyl acetate. The combined organic fractions were dried _{over Na2SO4} ; concentrated and purified by flash column chromatography (15% ethyl acetate in hexanes) (82_Int-3) . Confirmed by 2D NMR (ROE analysis). MS (ES): 328 m/z, LCMS purity: 99.11%, 1H 400 MHz, DMSO-d6: δ 1.654 (s, 6H), 3.258 (s, 3H), 3.758 (s, 3H), 4.797 (s, 2H), 5.394 (s, 2H), 7.258 (d, J= 8 Hz, 2H), 7.478 (d, J=8 Hz, 2H), 7.903 (s, 1H). Example 10A : Preparation of methyl 1-(4-( cyanomethyl ) benzyl )-5-( methoxymethyl )-1H- pyrazole -4- carboxylate (85_Int-3) Step -1. Synthesis of methyl 1-(4-( cyanomethyl ) benzyl )-5-( methoxymethyl )-1H- pyrazole -4- carboxylate (85_Int-3)

A stirred solution of methyl 5-(methoxymethyl)-1H-pyrazole-4-carboxylate (82_Int-2) (1.0 g, 5.87 mmol, 1.0 eq) in DMF (10 mL, 10 V) was prepared and NaHMDS (1M in THF) (5.8 mL, 5.87 mmol, 1.0 eq) was added at 0 °C and stirred for 30 min. Then 2-(4-(bromomethyl)phenyl)acetonitrile (1.4 g, 7.05 mmol, 1.2 eq) was added to the RM at 0 °C and stirred for additional 16 h at RT. After the reaction was complete, the RM was quenched into water and extracted with ethyl acetate. The combined organic fractions were dried _{over Na2SO4} ; concentrated and purified by flash column chromatography (25-30% ethyl acetate in hexanes) (85_Int-3) . Confirmed by 2D NMR (ROE analysis). MS (ES): 300 m/z, 1H 400 MHz, DMSO-d6: δ 3.233 (s, 3H), 3.745 (s, 3H), 4.029 (s, 2H), 4.514 (s, 2H), 5.352 (s, 2H), 7.356-7.301 (m, 4H), 8.046 (s, 1H). Example 11A : Preparation of 1-(4-(2-( dimethylamino )-2 -oxoethyl ) benzyl )-1H- pyrazole -4- carboxylic acid ethyl ester (33_Int-3) Step -1. Synthesis of 2-(4-( bromomethyl ) phenyl )-N- methylacetamide (33_Int-2)

A stirred solution of 2-(4-(bromomethyl)phenyl)acetic acid (33_Int-1) (1.0 g, 4.36 mmol, 1.0eq) in toluene (10 mL, 10V) was prepared. _SOCl2 (0.54 g, 4.53 mmol, 1.04eq) and DMF (0.051 g, 0.69 mmol, 0.16eq) were added at RT and stirred at 85 °C for 3 h. The RM was cooled to 0 °C and then dimethylamine (2M in THF) (2 mL, 2V) was added. The RM was brought to RT and stirred for 6 h. After completion of the reaction, the RM was quenched into saturated NaHCO3 solution and extracted with ethyl _acetate . The combined organic fractions were dried over _Na2SO4 ; concentrated to give compound (33_Int-2) . MS (ES): 256.6 m/z [M]+ 258.1.m/z [M+2]+, LCMS purity: 98.96%, 1H NMR (400 MHz, DMSO-d6) δ 2.826 (s, 3H), 2.997 (s, 3H), 3.694 (s, 2H), 4.706 (s, 3H), 7.210 (d, J=8 Hz, 2H), 7.385 (d, J=8 Hz, 2H). Step -2. Synthesis of ethyl 1-(4-(2-( dimethylamino )-2- oxoethyl ) benzyl )-1H- pyrazole -4- carboxylate (33_Int-3) .

Prepare a stirring solution of ethyl 1H-pyrazole-4-carboxylate (0.3 g, 2.14 mmol, 1.0 eq) in acetone (6 mL, 20 V). 2-(4-(Bromomethyl)phenyl)-N,N-dimethylacetamide (33_Int-2) (0.657 g, 2.56 mmol, 1.2 eq) and Cs ₂ CO ₃ (1.67 g, 5.13 mmol, 2.4 eq) are added at RT. The RM is then heated to 65 °C and stirred for 16 h. After the reaction is complete, the RM is cooled to RT. The RM is filtered to remove Cs ₂ CO ₃ and washed with ethyl acetate. The solid residue was discarded and the filtrate was concentrated and purified by column chromatography (25-30% ethyl acetate in hexanes) to give compound (33_Int-3) MS (ES): 316.3 m/z [M+1]+, LCMS purity: 100%, 1H NMR (400 MHz, DMSO-d6) δ 1.252 (t, J= 2.8 Hz, 3H), 2.802 (s, 3H), 2.974 (s, 3H), 3.396-3.295 (m, 2H), 4.188 (s, 2H), 5.324 (s, 2H), 7.189 (s, 4H), 7.852 (s, 1H), 8.466 (s, 1H). Preparation Example 1 of the Final Compound : Preparation of N-(4- carbamimidoylbenzyl )-1-(4-(2- cyanopropan -2- yl ) benzyl )-1H -pyrazole -4- carboxamide (7)

In a 3000 mL 4N RBF attached to a mechanical stirrer and condenser was poured ethyl 1-(4-(2-cyanopropan-2-yl)benzyl)-1H-pyrazole-4-carboxylate (7-Int5) (120 g, 403 mmol, 1.0 eq), 4-(aminomethyl)benzamidamide dihydrochloride (134.4 g, 605 mmol, 1.5 eq) and toluene (1200 mL, 10V) at RT. DIPEA (130.14 g, 1008 mmol, 2.5 eq) was added to the RM. The RM was cooled to 0-5°C and stirred for 20 min. TMA (2M in toluene) (605.3 mL, 121 mmol, 3.0 eq) was added dropwise to the RM. The RM was heated to 95 °C and stirred for 16 h. After completion of the reaction, the reaction mixture was cooled to RT. The RM was slowly quenched with DM water (151 mL) and evaporated to obtain a residue. The filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified three times by column chromatography using 60 to 120 mesh size silica. The product was eluted with 10% MeOH in DCM to obtain a solid, which was dissolved in 4V methanol and stirred at 70 °C for 2 h. EtOAc was added at 60 °C until a cloudy solution was observed. The solution was slowly cooled to RT and stirred for 16 h until a solid precipitate was obtained, which was collected by filtration. The filtrate was evaporated and purified by column chromatography to give compound (7). MS (ES): 473.87 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 95.07% ¹ H NMR (400 MHz, DMSO-d6) δ 1.682 (s, 6H), 4.470 (s, 2H), 5.362 (s, 2H), 7.326 (d, J=8.4 Hz, 2H), 7.515-7.419 (m, 4H), 7.772 (d, J=8.4 Hz, 2H), 7.939 (s, 1H), 8.316 (s, 1H), 8.865-8.835 (m, 1H), 9.123 (s, 2H), 9.303 (s, 2H). Example 2 : N-(4- carbamimidoylbenzyl )-1-(4-(2- cyanoethyl ) benzyl )-1H- pyrazole -4- carboxamide (3) Step -1. Synthesis of ethyl 1-(4- iodobenzyl )-1H- pyrazole -4- carboxylate (3-Int-2) .

A stirred solution of 1H-pyrazole-4-carboxylate (0.5 g, 3.56 mmol, 1.0 eq) in DMF (5 mL, 10 V) was prepared and NaH (60% in mineral oil) (0.213-g, 5.35 mmol, 1.5 eq) was added at 0 °C. The RM was stirred at 0 °C for 30 min. 1-(Bromomethyl)-4-iodobenzene (1.05 g, 3.56 mmol, 1.0 eq) was added at 0 °C and the RM was brought to RT and stirred for 2 h. After the reaction was complete, the RM was quenched into water and extracted with ethyl acetate. The combined organic fractions were dried _{over Na2SO4} ; concentrated and purified by flash column chromatography (12-15% ethyl acetate in hexanes) to give compound (3-Int-2) MS (ES): 357.1.m/z [M+H]+, LCMS purity: 100%, 1H 400 MHz, DMSO-d6: δ 1.25 (t, J = 7.20 Hz, 3H), 4.20 (d, J = 7.20 Hz, 2H), 5.33 (s, 2H), 7.066 (d, J = 7.60 Hz, 2H), 7.715 (d, J = 8.00 Hz, 2H), 7.87 (s, 1H), 8.47 (s, 1H). Step -2. Synthesis of (E)-ethyl 1-(4-(2- cyanovinyl ) benzyl )-1H- pyrazole -4- carboxylate (3_Int-3) .

Preparation of 1-(4-iodobenzyl)-1H-pyrazole-4-carboxylic acid ethyl ester (3_Int-2) (0.55 g, 1.54 mmol, 1.0 eq) and 1,4-dimethoxybenzyl To a stirred solution of 2-nitropropane (11 mL, 20V) was added acrylonitrile (0.08 g, 1.54 mmol, 1eq) and TEA (0.390 g, 3.86 mmol, 2.5eq) at RT. N2 was purged in the RM at RT for 10 min. Palladium diacetate (0.034 g, 0.15momol, 0.1eq) and JohnPhos (0.092 g, 0.30momol, 0.2eq) were added and the RM was heated to 110°C and stirred for 3h. After the reaction was complete, the RM was quenched in water and extracted with ethyl acetate. The combined organic fractions were dried _{over Na2SO4} ; concentrated and purified by flash column chromatography (22-25% ethyl acetate in hexanes) to give compound (3-Int-3) MS (ES): 282.30.m/z [M+H]+, LCMS purity: 99.41%, 1H 400 MHz, DMSO-d6: δ 1.29 (d, J = 5.6 Hz, 3H), 2.231 (d, J = 5.6 Hz, 2H), 5.441 (d, J = 7.2 Hz, 2H), 5.92 (d, J = 12 Hz, 1H), 7.444-7.328 (m, Hz, 3H), 7.799 (d, J = 5.6 Hz, 2H), 7.907 (s, Hz, 1H), 8.533 (s, 1H). Step -3. Synthesis of ethyl 1-(4-(2- cyanoethyl ) benzyl )-1H- pyrazole -4- carboxylate (3-Int-4) .

A stirred solution of (E)-1-(4-(2-cyanovinyl)benzyl)-1H-pyrazole-4-carboxylate (3-Int-3) (0.32 g, 1.13 mmol, 1.0 eq) in THF (6 mL, 20 V) was prepared and 5% Pd/C was added at RT. The RM was stirred under H2 atmosphere at RT for 8 h. The RM was filtered through a celite bed washed with ethyl acetate. The filtrate was concentrated and then purified by column chromatography (20-22% EtOAc in hexanes) to give compound (3-Int-4). MS (ES): 284.30.m/z [M+H]+, LCMS purity: 65.00%, 1H 400 MHz, DMSO-d6: δ 1.32 (t, J = 6.8 Hz, 3H), 2.81 (d, J = 6.00 Hz, 2H), 2.86 (d, J = 6.00 Hz, 2H), 4.22 (d, J = 6.80 Hz, 2H), 5.36 (s, 2H), 7.27 (d, J = 7.60 Hz, 2H), 7.66 (d, J = 8.80 Hz, 2H), 7.87 (s, 1H), 8.50 (s, 1H). Step -4. Synthesis of N-(4- carbamimidoylbenzyl )-1-(4-(2- cyanoethyl ) benzyl )-1H- pyrazole -4- carboxamide (3) .

A stirred solution of ethyl 1-(4-(2-cyanoethyl)benzyl)-1H-pyrazole-4-carboxylate (3_Int-4) (0.2 g, 7.05 mmol, 1.0 eq) in toluene (2 mL, 10 V) was prepared and 4-(aminomethyl)benzamididine dihydrochloride (0.15 g, 10.58 mmol, 1.5 eq) and DIPEA (0.23 g, 17.64 mmol, 2.5 eq) were added at RT. After cooling to 0 °C, TMA (2.0 M in toluene) (1 mL, 2.11 mmol, 3.0 eq) was added and the RM was heated to 100 °C and stirred for 16 h. After the reaction was complete, the RM was quenched by 1 V of water and evaporated. The residue was washed with 20% methanol in dichloromethane and filtered. The filtrate was concentrated and purified by Prep HPLC ((A) 0.1% TFA in water (B) 100% MeCN)) to give compound (3). MS (ES): 387.29.m/z [M+H]+, LCMS purity: 100%, HPLC purity: 100% 1H 400 MHz, DMSO-d6: δ 2.80 (d, J = 6.00 Hz, 2H), 2.84 (d, J = 6.00 Hz, 2H), 4.48 (d, J = 5.60 Hz, 2H), 5.33 (s, 2H), 7.25 (q, J = 27.20 Hz, 3H), 7.49 (d, J = 8.00 Hz, 1H), 7.74 (d, J = 8.00 Hz, 1H), 0.00 (s, 1H), 8.27 (s, 1H), 8.78 (d, J = 6.00 Hz, 1H), 8.95 (s, 2H), 9.25 (s, 2H). Example 3 : N-(4 -Carboxamidinobenzyl )-1-(4-(2- cyanoethyl ) benzyl )-1H- imidazole -4 -carboxamide (4) Step -1. Synthesis of ethyl 1-(4- iodobenzyl )-1H- imidazole -4- carboxylate (4-Int-2) .

A stirred solution of 1H-imidazole-4-carboxylic acid ethyl ester (0.5 g, 3.56 mmol, 1.0 eq) in DMF (5 mL, 10 V) was prepared and NaHMDS (1M in THF) (3.5 mL, 3.56 mmol, 1.0 eq) was added at 0 °C and stirred for 30 min. 1-(Bromomethyl)-4-iodobenzene (0.105 g, 3.56 mmol, 1.0 eq) was added to the RM at 0 °C, brought to RT and stirred for 2 h. After the reaction was complete, the RM was quenched in water and extracted with ethyl acetate. The combined organic fractions were dried over Na ₂ SO ₄ ; concentrated and then purified by flash column chromatography (35-38% ethyl acetate in hexanes) to give compound (4_Int-2). MS (ES): 357.20.m/z [M+H]+, LCMS purity: 99.05%, 1H 400 MHz, DMSO-d6: δ 1.24 (t, J = 7.2 Hz, 3H), 4.19 (q, J = 7.2 Hz, 2H), 5.20 (s, 2H), 7.12 (d, J = 8.00 Hz, 2H), 7.74 (d, J = 8.00 Hz, 2H), 7.88 (s, 1H), 7.94 (s, 1H). Step -2. Synthesis of (E)-ethyl 1-(4-(2- cyanovinyl ) benzyl )-1H- pyrazole -4- carboxylate (4-Int-3) .

Preparation of 1-(4-iodobenzyl)-1H-imidazole-4-carboxylic acid ethyl ester (4_Int-2) (0.7 g, 1.96 mmol, 1.0 eq) and 1,4-dimethoxybenzyl To a stirred solution of 1,4-dioxane (14 mL, 20V) was added acrylonitrile (0.104 g, 1.96 mmol, 1eq) and TEA (0.497 g, 4.91 mmol, 2.5eq) at RT. N2 was purged in the RM at RT for 10 min. Palladium diacetate (0.044 g, 0.19 mmol, 0.1eq) and JohnPhos (0.117 g, 0.39momol, 0.2eq) were added to the RM. The RM was heated to 110°C and stirred for 3h. The RM was quenched in water and extracted with ethyl _acetate . The combined organic fractions were dried over _Na2SO4 ; concentrated and then purified by flash column chromatography (22-25% ethyl acetate in hexane) to give compound (3_Int-3). MS (ES): 282.28.m/z [M+H]+, LCMS purity: 89.04%, 1H 400 MHz, DMSO-d6: δ 1.23 (t, J = 7.2 Hz, 3H), 4.21 (q, J = 5.60 Hz, 2H), 5.22 (s, 2H), 6.49 (d, J = 16.80 Hz, 1H), 7.41 (t, J = 6.8 Hz, 3H), 7.66 (d, J = 6 Hz, 1H), 7.82 (d, J = 7.60 Hz, 2H), 7.99 (s, 1H). Step -3. Synthesis of ethyl 1-(4-(2- cyanoethyl ) benzyl )-1H- imidazole -4- carboxylate (4-Int-4) .

A stirring solution of (E)-ethyl 1-(4-(2-cyanovinyl)benzyl)-1H-imidazole-4-carboxylate (4-Int-3) (0.4 g, 1.42 mmol, 1.0 eq) in THF (5 mL, 12 V) and ethanol (5 mL, 12 V) was prepared and 5% Pd/C was added at RT. The RM was stirred in hydrogen atmosphere at RT for 16 h. After completion of the reaction, the RM was filtered through a celite bed washed with ethyl acetate. The filtrate was concentrated and then purified by column chromatography (38-40% EtOAc in hexanes) to give compound (3-Int-4). MS (ES): 284.22.m/z [M+H]+, LCMS purity: 69.16.

Step-4. N-(4-Carbamimidoylbenzyl)-1-(4-(2-cyanoethyl)benzyl)-1H-pyrazole-4-carboxamide (4) is prepared from ethyl 1-(4-(2-cyanoethyl)benzyl)-1H-imidazole-4-carboxylate (4_Int-4) and 4-(aminomethyl)benzamimidone dihydrochloride in a similar manner to that described in Example-2 (4). MS (ES): 387.30.m/z [M+H]+, LCMS purity: 100%, HPLC purity: 100% 1H 400 MHz, DMSO-d6: δ 2.80 (d, J = 6.40 Hz, 2H), 2.85 (d, J = 6.00 Hz, 2H), 4.47 (d, J = 5.60 Hz, 2H), 5.23 (s, 2H), 7.30 (s, 3H), 7.46 (d, J = 28.00 Hz, 2H), 7.74 (t, J = 8 Hz, 3H), 8.00 (s, 1H), 8.74 (s, 1H), 8.95 (s, 2H), 9.24 (s, 2H). Example 4 : Preparation of N-(4- carbamimidoyl -3- fluorobenzyl )-1-(4-(2- cyanopropan -2- yl ) benzyl )-1H- pyrazole -4- carboxamide (43)

N-(4-Carbamimidoyl-3-fluorobenzyl)-1-(4-(2-cyanopropan-2-yl)benzyl)-1H-pyrazole-4-carboxamide (301) was prepared from ethyl 1-(4-(2-cyanopropan-2-yl)benzyl)-1H-pyrazole-4-carboxylate (7-Int-5) and 4-(aminomethyl)-2-fluorobenzamidine dihydrochloride (43-Int-5) in a similar manner as described in Example 2 (3). MS (ES): 419.20.m/z [M+H]+, LCMS purity: 100%, HPLC purity: 10% ¹ H NMR (400 MHz, DMSO-d6) δ 1.667 (s, 6H), 2.504 (s, 2H), 4.771 (d, J=4.2 Hz, 2H), 5.368 (s, 2H), 7.343-7.324 (m, 4H), 7.511 (d, J=8 Hz, 2H), 7.625 (t, J=8 Hz, 1H), 7.919 (s, 1H), 8.295 (s, 1H), 8.795 (t, J=6 Hz, 1H), 9.184 (s, 2H), 9.369 (s, 2H). Example 5 : N-(4- carbamimidoylbenzyl )-1-(4-( cyanomethyl ) phenethyl )-1H -pyrazole -4- carboxamide (12) Step -1. Synthesis of ethyl 1-(4- methylphenethyl )-1H- pyrazole -4- carboxylate (12-Int-2) .

A stirred solution of ethyl 1H-pyrazole-4-carboxylate (2 g, 14.28 mmol, 1.0 eq) in acetone (20 mL, 10 V) was prepared and 1-(2-bromoethyl)-4-methylbenzene (304 g, 17.14 mmol, 1.2 eq) and Cs ₂ CO ₃ (11.14 g, 34.28 mmol, 2.4 eq) were added at RT. The RM was heated to 60-65 °C, stirred for 16 h, and cooled to RT. The RM was filtered, washed with EtOAc, and purified by column chromatography (12-15% ethyl acetate in hexanes) to give compound (12_Int-2) MS (ES): 259.23.m/z [M+H]+. Step -2. Synthesis of ethyl 1-(4-( bromomethyl ) phenethyl )-1H- pyrazole -4- carboxylate (12-Int-3) .

A stirred solution of ethyl 1-(4-methylphenethyl)-1H-pyrazole-4-carboxylate (12_Int-2) (1.7 g, 6.58 mmol, 1.0 eq) in carbon tetrachloride (34 mL, 20 V) was prepared and benzoyl peroxide (0.159 g, 0.65 mmol, 0.1 eq) and N-bromosuccinimide (1.40 g, 7.89 mmol, 1.2 eq) were added at RT. The RM was stirred at RT for ₄ h, quenched into water and extracted with DCM. The combined organic fractions were dried over _Na2SO4 ; concentrated and purified by flash column chromatography (10-12% ethyl acetate in hexanes) to give compound (12_Int-3). MS (ES): 337.4.m/z [M]+ 339.4 [M+2]+, LCMS purity: 75.91%, ¹ H NMR (400 MHz, DMSO-d6) δ 1.239 (t, J=7.2 Hz, 3H), 3.111 (t, J=7.2 Hz, 2H), 4.179 (q, J=7.2 Hz, 2H), 4.385 (t, J=7.2 Hz, 2H), 4.664 (s, 2H), 7.148-7.127 (m, 2H), 7.350-7.330 (m, 2H), 7.841 (s, 1H), 8.220 (s, 1H). Step -3. Synthesis of ethyl 1-(4-( cyanomethyl ) phenethyl )-1H- pyrazole -4- carboxylate (12_Int-4) .

A stirred solution of ethyl 1-(4-(bromomethyl)phenethyl)-1H-pyrazole-4-carboxylate (12_Int-3) (0.750 g, 2.22 mmol, 1.0 eq) in MeCN (6. mL, 20 V) was prepared at RT. Tetrabutylammonium cyanide (2.38 g, 8.89 mmol, 4.0 eq) was added and the RM was stirred at RT for 16 h. After completion of the reaction, the RM was quenched in water and extracted with ethyl _acetate . The combined organic fractions were dried over _Na2SO4 ; concentrated and purified by flash column chromatography (7-10% ethyl acetate in hexanes) to give compound (12_Int-4). MS (ES): 284.28 m/z [M+H]+, LCMS purity: 93.34%, ¹ H NMR (400 MHz, DMSO-d6) δ 1.239 (t, J=7.2 Hz, 3H), 3.112 (t, J=7.2 Hz, 2H), 3.975 (s, 2H), 4.188 (q, J=7.2 Hz, 2H), 4.383 (t, J=7.2 Hz, 2H), 7.164 (d, J=8 Hz, 2H), 7.238 (d, J=8 Hz, 2H), 7.839 (s, 1H), 8.216 (s, 1H). Step -4. Synthesis of N-(4- carbamimidoylbenzyl )-1-(4-( cyanomethyl ) phenethyl )-1H -pyrazole -4- carboxamide (12)

The title compound was prepared from ethyl 1-(4-(cyanomethyl)phenethyl)-1H-pyrazole-4-carboxylate (12-Int-4) and 4-(aminomethyl)benzamidine dihydrochloride in a manner similar to that described in Example 2. MS (ES): 387.37 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 95.02% ¹ H NMR (400 MHz, DMSO-d6) δ 3.110 (t, J=7.2 Hz, 2H), 3.979 (s, 2H), 4.367 (t, J=7.2 Hz, 2H), 4.771 (d, J=6 Hz, 2H), 7.182 (d, J=8 Hz, 2H), 7.244 (d, J=8 Hz, 2H), 7.485 (d, J=8 Hz, 2H), 7.751 (d, J=8 Hz, 2H), 7.900 (s, 1H), 8.086 (s, 1H), 8.720 (t, J = 6 Hz, 1H), 8.974 (s, 2H), 9.248 (s, 2H). Example 6 : Preparation of 2-(4-((4-((4 -carbamimidoylbenzyl ) aminomethyl )-1H- pyrazol -1- yl ) methyl ) phenyl ) acetic acid (16) Step -1. Synthesis of 2-(4-((4-( ethoxycarbonyl )-1H- pyrazol -1- yl ) methyl ) phenyl ) acetic acid (16-Int-2) .

A stirred solution of 1H-pyrazole-4-carboxylate (0.5 g, 3.56 mmol, 1.0 eq) in DMF (5 mL, 10 V) was prepared and NaH (60% in mineral oil) (0.214 g, 5.35 mmol, 1.5 eq) was added at 0 °C. The RM was stirred at 0 °C for 30 min. 2-(4-(Bromomethyl)phenyl)acetic acid (0.980 g, 4.28 mmol, 1.2 eq) was added to the RM at 0 °C and stirred at RT for 16 h. After the reaction was complete, the RM was quenched into water and extracted with ethyl acetate. The combined organic fractions were dried _{over Na2SO4} ; concentrated and purified by flash column chromatography (5-7% methanol in DCM) to give compound (16-Int-2) MS (ES): 289.3 m/z [M+H]+, LCMS purity: 47.20%, ^1H NMR (400 MHz, DMSO-d6) δ 1.252 (t, J=7.2 Hz, 3H), 3.542 (s, 2H), 4.201 (q, J=7.2 Hz, 2H), 5.335 (s, 2H), 7.222 (s, 4H), 7.853 (s, 1H), 8.451 (s, 1H), 12.321 (s, 1H).

Step-2. 2-(4-((4-((4-Carbamimidoylbenzyl)aminocarbonyl)-1H-pyrazol-1-yl)methyl)phenyl)acetic acid (16) is prepared from 2-(4-((4-(ethoxycarbonyl)-1H-pyrazol-1-yl)methyl)phenyl)acetic acid (16-Int-2) and 4-(aminomethyl)benzamidine dihydrochloride in a similar manner to that described in Example-2 (16). MS (ES): 392.50 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 99.29% ¹ H NMR (400 MHz, DMSO-d6) δ 1.841 (s, 2H), 3.503 (s, 2H), 4.785 (s, 2H), 7.073-7.051 (m, 4H), 7.297 (d, J=8 Hz, 2H), 7.504 (d, J=8 Hz, 2H), 7.794 (s, 1H), 8.065 (s, 1H). Example 7 : Preparation of 1-(4-(2- amino -2- oxoethyl ) benzyl )-N-(4- carbamimidoylbenzyl )-1H- pyrazole -4- carboxamide (18) Step -1. Synthesis of 2-(4-( bromomethyl ) phenyl ) acetamide (18-Int-2) .

A stirred solution of 2-(4-(bromomethyl)phenyl)acetic acid (0.5 g, 2.18 mmol, 1.0 eq) in toluene (5 mL, 10 V) was prepared and SOCl2 (0.270 g, 2.26 mmol, 1.04 eq) and DMF (0.025 g, 0.34 mmol, 0.16 eq) were added at RT. The RM was heated at 80-85 °C for 3 h followed by a NH3 purge at 0 °C for 3 h. The RM was filtered and the solid was washed with EtOAc to give compound (18_Int-2). MS (ES): 228.30 m/z [M]+, 230.12 m/z [M+2]+. Step -2. Synthesis of ethyl 1-(4-(2- amino -2- oxoethyl ) benzyl )-1H- pyrazole -4- carboxylate (18_Int-3) .

A stirred solution of 1H-pyrazole-4-carboxylate (18_Int-2) (0.5 g, 3.56 mmol, 1.0 eq) in DMF (5 mL, 10 V) was prepared and NaH (60% in mineral oil) (0.214 g, 5.35 mmol, 1.5 eq) was added at 0 °C. The RM was stirred at 0 °C for 30 min. 2-(4-(Bromomethyl)phenyl)acetamide (0.976 g, 4.28 mmol, 1.2 eq) was added to the RM at 0 °C and stirred at RT for 16 h. After the reaction was complete, the RM was quenched into water and extracted with ethyl acetate. The combined organic fractions were dried _{over Na2SO4} ; concentrated and purified by flash column chromatography (40-42% ethyl acetate in hexanes) to give compound (18_Int-3). MS (ES): 288.23.m/z [M+H]+, LCMS purity: 97.01%, ^1H NMR (400 MHz, DMSO-d6) δ 1.244 (t, J=7.2 Hz, 3H), 3.231 (s, 3H), 4.201 (q, J=7.2 Hz, 2H), 4.210 (s, 2H), 5.320 (s, 2H), 7.210 (s, 4H), 7.917-7.848 (m, 2H), 8.436 (s, 1H). Step -3. Synthesis of 1-(4-(2- amino -2- oxoethyl ) benzyl )-N-(4- carbamimidoylbenzyl )-1H- pyrazole -4- carboxamide (18)

The title compound was prepared from ethyl 1-(4-(2-amino-2-oxoethyl)benzyl)-1H-pyrazole-4-carboxylate (18-Int-3) and 4-(aminomethyl)benzamididine dihydrochloride in a manner similar to that described in Example 2. MS (ES): 391.32.m/z [M+H]+, LCMS purity: 99.23%, HPLC purity: 98.73% ¹ H NMR (400 MHz, DMSO-d6) δ 3.321 (d, J=3.2 Hz, 2H), 4.479 (d, J=6 Hz, 2H), 5.313 (s, 2H), 6.489 (s, 1H), 7.247-7.193 (m, 4H), 7.505-7.449 (m, 3H), 7.748 (d, J=8 Hz, 1H), 7.902 (s, 1H), 8.253 (s, 1H), 8.752 (t, J=6 Hz, 1H), 8.967 (s, 2H), 9.244 (s, 2H). Example 8 : Preparation of N-(4- carbamimidoylbenzyl )-1-(4-(2-( methylamino )-2- oxoethyl ) benzyl )-1H- pyrazole -4- carboxamide (19) Step -1. Synthesis of 2-(4-( bromomethyl ) phenyl )-N -methylacetamide (19-Int-2) .

A stirred solution of 2-(4-(bromomethyl)phenyl)acetic acid (1.0 g, 4.36 mmol, 1.0 eq) in toluene (5 mL, 10V) was prepared at RT. SOCl2 (0.54 g, 4.53 mmol, 1.04 eq) and DMF (0.051 g, 0.69 mmol, 0.16 eq) were added at RT and then heated to 80-85 °C for 3 h. The solution was cooled to 0 and methylamine (2 mL) was added to the RM and stirred at RT for 6 h. The RM was filtered and the solid was washed with EtOAc to give compound (19-Int-2). MS (ES): 242.1.m/z [M]+ 244.1.m/z [M+2]+, LCMS purity: 96.6%, ¹ H NMR (400 MHz, DMSO-d6) δ 3.389 (s, 2H), 4.726 (s, 2H), 7.245 (d, J=8 Hz, 2H), 7.344 (d, J=8 Hz, 2H), 8.019 (s, 1H). Step -2. Synthesis of ethyl 1-(4-(2-( methylamino )-2 -oxoethyl ) benzyl )-1H -pyrazole -4- carboxylate (19_Int-3) .

To a stirred solution of ethyl 1H-pyrazole-4-carboxylate (0.5 g, 3.56 mmol, 1.0 eq) in acetone (10 mL, 20 V) was prepared 2-(4-(bromomethyl)phenyl)-N-methylacetamide (19_Int-2) (1.03 g, 4.28 mmol, 1.2 eq) and S ₂ CO ₃ (2.78 g, 8.56 mmol, 2.4 eq) were added at RT. The RM was heated to 60-65 °C and stirred for 16 h. After the reaction was complete, the RM was cooled to RT. The RM was filtered and washed with EtOAc. The solid residue was discarded and the filtrate was concentrated and purified by column chromatography (12-15% ethyl acetate in hexanes) to give compound (19-Int-3) MS (ES): 302.40.m/z [M+H]+, LCMS purity: 86.48%, ¹ H NMR (400 MHz, DMSO-d6) δ 1.658 (s, 6H), 4.556 (s, 2H), 5.366 (s, 2H), 7.331 (d, J=6 Hz, 2H), 7.505 (d, J=8 Hz, 4H), 7.983-7.934 (m, 2H), 8.314 (s, 1H), 8.595 (s, 1H), 8.813 (s, 1H).

Step-3. N-(4-Carbamimidoylbenzyl)-1-(4-(2-(methylamino)-2-oxoethyl)benzyl)-1H-pyrazole-4-carboxamide (19) was prepared from ethyl 1-(4-(2-(methylamino)-2-oxoethyl)benzyl)-1H-pyrazole-4-carboxylate (19-Int-3) and 4-(aminomethyl)benzamimidoyl dihydrochloride in a similar manner as described in Example 2. MS (ES): 405.55.m/z [M+H]+, LCMS purity: 100%, HPLC purity: 100% ¹ H NMR (400 MHz, DMSO-d6) δ 2.545 (d, J=4.8 Hz, 3H), 3.361 (s, 2H), 4.476 (d, J=6 Hz, 2H), 5.306 (s, 2H), 7.134-7.186 (m, 4H), 7.492 (d, J=8 Hz, 2H), 7.743 (d, J=8 Hz, 2H), 7.916-7.896 (m, 2H), 8.246 (s, 1H), 8.748 (t, J=6 Hz, 1H), 8.896 (s, 2H), 9.234 (s, 2H). Example 9 : Preparation of N-(4 -carbamimidoylbenzyl )-1-(4-((3- cyano -1H- pyrrol -1- yl ) methyl ) benzyl )-1H- pyrazole -4 -carboxamide (26 )

N-(4-Carbamimidoylbenzyl)-1-(4-((3-cyano-1H-pyrrol-1-yl)methyl)benzyl)-1H-pyrazole-4-carboxamide (26) was prepared from ethyl 1-(4-((3-cyano-1H-pyrrol-1-yl)methyl)benzyl)-1H-pyrazole-4-carboxylate (6-Int-1) and 4-(aminomethyl)benzamimidoyl dihydrochloride in a manner similar to that described in the Examples. MS (ES): 43841.m/z [M+H]+, LCMS purity: 100%, HPLC purity: 100% ¹ H NMR (400 MHz, DMSO-d6) δ 4.482 (d, J=6 Hz, 2H), 5.156 (s, 2H), 5.335 (s, 2H), 6.467 (s, 2H), 7.001 (s, 1H), 7.254-7.250 (m, 4H), 7.496 (d, J=8 Hz, 2H), 7.711 (s, 1H), 7.754 (d, J=8 Hz, 2H), 7.909 (s, 1H), 8.270 (s, 1H), 8.772 (t, J=6 Hz, 1H), 9.012 (s, 2H), 9.258 (s, 2H). Example 10 : Preparation of N-(4- carbamimidoylbenzyl )-1-(4-(1- cyanoethyl ) benzyl )-1H -pyrazole -4- carboxamide (5) Step -1. Synthesis of ethyl 1-(4-(1- cyanoethyl ) benzyl )-1H- pyrazole -4- carboxylate (5-Int-1) .

A stirred solution of ethyl 1-(4-(cyanomethyl)benzyl)-1H-pyrazole-4-carboxylate (1_int-4) (0.3 g, 1.11 mmol, 1.0 eq) in THF (6 mL, 20 V) was prepared and LiHMDS (1M in hexanes) (1.23 mL, 1.22 mmol, 1.1 eq) was added at -70 °C. The RM was stirred at -70 °C for 30 min. Methyl iodide (0.2 g, 1.44 mmol, 1.3 eq) was added to the RM at -70 °C. The RM was warmed to RT and stirred for 16 h. After the reaction was complete, the RM was quenched into water and extracted with ethyl acetate. The combined organic fractions were dried _{over Na2SO4} , concentrated and purified by flash column chromatography (12-15% ethyl acetate in hexanes) to give compound (5_Int-1). MS (ES): 284.28 m/z [M+H]+, LCMS purity: 46.39%.

Step-2. N-(4-amidinobenzyl)-1-(4-(1-cyanoethyl)benzyl)-1H-pyrazole-4-carboxamide (45) was prepared from 1-(4-(1-cyanoethyl)benzyl)-1H-pyrazole-4-carboxylic acid ethyl ester (5-Int-1) in a similar manner to that described in Example-2. MS (ES): 387.33.m/z [M+H]+, LCMS purity: 94.00%. Example 11 : Preparation of N-(4- amidino -2- fluorobenzyl )-1-(4-(2- cyanopropan -2- yl ) benzyl )-1H- pyrazole -4- carboxamide ( 45)

N-(4-Carbamimidoyl-2-fluorobenzyl)-1-(4-(2-cyanopropan-2-yl)benzyl)-1H-pyrazole-4-carboxamide (45) was prepared from ethyl 1-(4-(2-cyanopropan-2-yl)benzyl)-1H-pyrazole-4-carboxylate (4-Int-5) and 4-(aminomethyl)-3-fluorobenzamidine dihydrochloride (45-Int-5) in a similar manner as described in Example-2. MS (ES): 419.4.m/z [M+H]+, LCMS purity: 100%, HPLC purity: 100% ¹ H NMR (400 MHz, DMSO-d6) δ 1.664 (s, 6H), 4.499 (d, J=6.4 Hz, 2H), 5.366 (s, 2H), 7.331 (d, J=8 Hz, 2H), 7.554-7.500 (m, 3H), 7.608 (d, J=8 Hz, 1H), 7.771 (d, J=8 Hz, 1H), 7.925 (s, 1H), 8.303 (s, 1H), 8.776 (tJ=8.8 Hz, 1H), 9.115 (s, 2H), 9.334 (s, 2H), ¹⁹ F NMR (400 MHz, DMSO-d6) δ -73-659 (1F), -116.871 (0.35F). Example 12 : Preparation of N-(4- carbamimidoylbenzyl )-1-(4-( cyanomethyl ) benzyl )-1H- pyrazole -4- carboxamide (1)

In a 3000 mL 4N RBF attached to a mechanical stirrer and condenser was poured ethyl 1-(4-(cyanomethyl)benzyl)-1H-pyrazole-4-carboxylate (1-Int4) (140 g, 517 mmol, 1.0 eq), 4-(aminomethyl)benzamidine dihydrochloride (127 g, 571 mmol, 1.1 eq), and toluene (1400 mL, 10 V) at RT. DIPEA (266.5 mL, 1559 mmol, 3.0 eq) was added to the RM. The RM was cooled to 0-5 °C and stirred for 20 min. TMA (2M in toluene) (649.8 mL, 1299 mmol, 2.5 eq) was added dropwise to the RM. The RM was heated to 95 °C and stirred for 16 h. The RM was cooled to RT, quenched slowly with DM water (151 mL), and evaporated to obtain a residue. The solid residue was washed with 50% MeOH in DCM (3x1000 mL) and the filtrate was discarded. The solid residue was washed again with 20% MeOH in DCM (2x2000 mL) and discarded. The filtrate was concentrated and purified by flash column chromatography (12-15% methanol in DCM) to give compound (1) MS (ES): 373.35 m/z [M+H]+, LCMS purity: 96.80%, HPLC purity: 95.11% ¹ H NMR (400 MHz, DMSO-d6) δ 3.805 (s, 2H), 4.455 (s, 2H), 5.314 (s, 2H), 7.304-7.289 (m, 4H), 7.469 (d, J=8 Hz 2H), 7.710 (d, J=8 Hz, 2H), 7.897 (s, 1H), 8.246 (s, 1H). Example 13 : Preparation of N-(4- carbamimidoylbenzyl )-1-(4-( hydroxymethyl ) benzyl )-1H -pyrazole -4- carboxamide (2) Step -1. Synthesis of ethyl 1-(4-( hydroxymethyl ) benzyl )-1H- pyrazole -4- carboxylate (2-Int-2)

A stirred solution of ethyl 1H-pyrazole-4-carboxylate (2_int-1) (0.5 g, 3.56 mmol, 1.0 eq) in acetone (5 mL, 10 V) was prepared and (4-(bromomethyl)phenyl)methanol (0.86 g, 4.28 mmol, 1.2 eq) and Cs ₂ CO ₃ (3.25 g, 9.98 mmol, 2.8 eq) were added at RT. The RM was heated to 60 °C and stirred for 3 h. After the reaction was complete, the RM was quenched in water and extracted with ethyl acetate. The combined organic fractions were dried over Na ₂ SO ₄ ; concentrated and then purified by flash column chromatography (12-15% ethyl acetate in hexanes) to give compound (2_Int-2). MS (ES): 261.30 m/z [M+H]+, LCMS purity: 97.45%, ¹ H NMR (400 MHz, DMSO-d6) δ 1.255 (s, 3H), 4.203 (d, J=6.4 Hz, 2H), 4.469 (s, 2H), 5.187 (s, 1H), 5.340 (s, 2H), 7.269 (m, 4H), 7.860 (s, 1H), 8.444 (s, 1H).

Step-2. N-(4-Carbamimidoylbenzyl)-1-(4-(hydroxymethyl)benzyl)-1H-pyrazole-4-carboxamide (2) was prepared from ethyl 1-(4-(hydroxymethyl)benzyl)-1H-pyrazole-4-carboxylate (2-Int-2) and 4-(aminomethyl)benzamimidone dihydrochloride in a similar manner to that described in Example-2. MS (ES): 364.28 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 95.74% ¹ H NMR (400 MHz, DMSO-d6) δ 4.469 (s, 1H), 5.199 (s, 1H), 5.321 (s, 2H), 7.282-7.223 (m, 4H), 7.485 (d, J=6.4 Hz, 2H), 7.741 (d, J=6.4 Hz, 2H), 7.904 (s, 1H), 8.251 (s, 1H), 8.776 (s, 1H), 8.995 (s, 2H), 9.250 (s, 2H), ¹⁹ F NMR (400 MHz, DMSO-d6) δ -73.575 (1F). Example 14 : Preparation of N-(4- carbamimidoylbenzyl )-1-(4-(2- cyanoprop -2- yl ) benzyl )-1H- imidazole -4- carboxamide (8) Step -1. Synthesis of ethyl 1-(4-(2- cyanopropan -2- yl ) benzyl )-1H- imidazole -4- carboxylate (8_Int-1)

A stirred solution of ethyl 1H-imidazole-4-carboxylate (0.45 g, 3.21 mmol, 1.0 eq) in DMF (4.5 mL, 10 V) was prepared and LiHMDS (1M in THF) (3.2 mL, 3.21 mmol, 1.0 eq) was added at -78 °C and stirred for 30 min. 2-(4-(Bromomethyl)phenyl)-2-methylpropionitrile (7_int-3) (0.91 g, 3.85 mmol, 1.2 eq) was added to the RM at -78 °C. The RM was cooled to RT and stirred at RT. After the reaction was complete, the RM was quenched into water and extracted with ethyl acetate. The combined organic fractions were dried _{over Na2SO4} ; concentrated and purified by flash column chromatography (1.2-1.5% methanol in DCM) to give compound (8_Int-1). MS (ES): 297.85 m/z [M+H]+, LCMS purity: 86.41%, ¹ H NMR (400 MHz, DMSO-d6) δ 1.246 (t, J=5.2 Hz, 3H), 1.649 (s, 6H), 4.220-4.185 (m, 2H), 5.256 (s, 2H), 7.381 (d, J=8 Hz, 2H), 7.524 (d, J=8 Hz, 2H), 7.895 (d, J=2 Hz, 1H), 7.952 (d, J=2 Hz, 1H).

Step-2. N-(4-Carbamimidoylbenzyl)-1-(4-(2-cyanopropan-2-yl)benzyl)-1H-imidazole-4-carboxamide (8) was prepared from ethyl 1-(4-(2-cyanopropan-2-yl)benzyl)-1H-imidazole-4-carboxylate (8_Int-1) and 4-(aminomethyl)benzamimidone dihydrochloride in a similar manner to that described in Example-2. MS (ES): 401.25 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 99.63% ¹ H NMR (400 MHz, DMSO-d6) δ 1.662 (s, 6H), 4.772 (d, J=6 Hz, 2H), 5.257 (s, 2H), 7.395 (d, J=8 Hz, 2H), 7.536-7.473 (m, 4H), 7.759-7.717 (m, 2H), 8.717 (s, 1H), 8.873 (s, 2H), 9.231 (s, 2H). Example 15 : Preparation of N-(4- carbamimidoylbenzyl )-1-(4-( cyanomethyl ) benzyl )-1H -imidazole -4- carboxamide (9) Step -1. Synthesis of 2-(4-( bromomethyl ) phenyl ) acetonitrile (9_Int-2)

A stirred solution of 2-(p-tolyl)acetonitrile (2 g, 15.24 mmol, 1.0 eq) and carbon tetrachloride (40 mL, 20 V) was prepared at RT. AIBN (0.12 g, 0.76 mmol, 0.1 eq) and N-bromosuccinimide (2.24 g, 1.67 mmol, 1.1 eq) were added at RT and the RM was heated to 90 °C and stirred for 16 h. After the reaction was complete, the RM was quenched in water and extracted with DCM. The combined organic fractions were dried over Na ₂ SO ₄ , concentrated and purified by flash column chromatography (7-10% ethyl acetate in hexanes) to give compound (9-Int-2). ¹ H NMR (400 MHz, DMSO-d6) δ 4.092 (s, 2H), 4.705 (s, 2H), 7.335 (d, J=8 Hz, 2H), 7.469 (d, J=8 Hz, 2H). Step -2. Synthesis of ethyl 1-(4-( cyanomethyl ) benzyl )-1H- imidazole -4- carboxylate (9_Int-3)

Prepare a stirred solution of ethyl 1H-imidazole-4-carboxylate (1.13 g, 8.09 mmol, 1.0 eq) in DMF (11.3 mL, 10 V) and add LiHMDS (1M in THF) (8.9 mL, 8.86 mmol, 1.1 eq) at -78 °C and stir for 30 min. Add 2-(4-(Bromomethyl)phenyl)acetonitrile (9-Int-2) (1.7 g, 8.09 mmol, 1.0 eq) to the RM at -78 °C. Bring the RM to RT and stir for 16 h. After the reaction is complete, quench the RM into water and extract with ethyl acetate. The combined organic fractions were dried _{over Na2SO4} , concentrated and purified by flash column chromatography (1.2-2.0% methanol in DCM) to give compound (9-Int-3). MS (ES): 270.23 m/z [M+H]+, LCMS purity: 77.67%, ^1H NMR (400 MHz, DMSO-d6) δ 1.239 (t, J=7.2 Hz, 3H), 4.020 (s, 2H), 4.186 (q, J=7.2 Hz, 2H), 5.240 (s, 2H), 7.348 (s, 4H), 7.877 (s, 1H), 7.929 (s, 1H).

Step-3. N-(4-Carbamimidoylbenzyl)-1-(4-(cyanomethyl)benzyl)-1H-imidazole-4-carboxamide (9) is prepared from ethyl 1-(4-(cyanomethyl)benzyl)-1H-imidazole-4-carboxylate (9_Int-3) and 4-(aminomethyl)benzamimidamide dihydrochloride in a similar manner to that described in Example-2. MS (ES): 373.29 m/z [M+H]+, LCMS purity: 98.06%, HPLC purity: 98.03% ¹ H NMR (400 MHz, DMSO-d6) δ 4.023 (s, 2H), 4.470 (s, 2H), 5.250 (s, 2H), 7.354 (s, 4H), 7.481 (d, J=6.0 Hz, 2H), 7.755-7.718 (m, 2H), 7.974 (s, 1H), 8.739 (s, 1H), 8.929 (s, 2H), 9.237 (s, 2H). Example 16 : N-(4 -carbamimidoylbenzyl )-3-(4-( cyanomethyl ) benzyl ) iso Preparation of oxazolidinone -5- carboxamide (10) Step -1. Synthesis of (E)-2-( p-tolyl ) acetaldehyde oxime (10_Int-2)

A stirred solution of 2-(p-tolyl)acetaldehyde (2.2 g, 16.39 mmol, 1.0eq) in DCM (44 mL, 20V) was prepared at RT. Hydroxylamine hydrochloride (2.3 g, 32.79 mmol, 2.0eq) and TEA (7 g, 68.86 mmol, 4.2eq) were added at 0°C. The RM was then brought to RT and stirred for 16 h. After completion of the reaction, the reaction mixture was diluted in DCM and washed with 1N HCl in saturated NaHCO3 and _brine solution. The combined organic fractions were dried over _Na2SO4 ; concentrated to give compound (10_Int-2). MS (ES): 150.10 m/z [M+H]+ Step -2. Synthesis of (Z)-N- hydroxy -2-( p-tolyl ) imidoacetyl chloride (10_Int-3)

A stirred solution of (E)-2-(p-tolyl)acetaldehyde oxime (10_Int-2) (4 g, 26.84 mmol, 1.0eq) in DMF (80 mL, 20V) was prepared and a solution of NCS (3.6 g, 26.84 mmol, 1.0eq) was added at 0°C. The RM was heated to 50°C and stirred for 4h. After completion of the reaction, the reaction mixture was evaporated. The remaining residue was quenched in _Na2S2O3 solution and extracted with ethyl acetate. The combined organic fractions were dried over _Na2SO4 ; concentrated to give compound (10_Int-3). MS (ES): 184.16m/z [M+H]+, LCMS purity: 60.43%. Step -3. 3-(4- methylbenzyl ) iso Synthesis of oxazole -5- carboxylic acid ethyl ester (10_Int-4)

A stirring solution of (Z)-N-hydroxy-2-(p-tolyl)imidoacetyl chloride (10_Int-3) (4 g, 21.78 mmol, 1.0 eq) in diethyl ether (80 mL, 20V) was prepared and TEA (3.3 g, 32.67 mmol, 1.5 eq) was added followed by ethyl propiolate (2.14 g, 21.78 mmol, 1.0 eq) at 0 °C and stirred for 4 h. After the reaction was complete, the RM was filtered. The filtrate was concentrated and then purified by flash column chromatography (1.2-2.0% ethyl acetate in hexanes) to give compound (10_Int-4). MS (ES): 246.24 m/z [M+H]+, LCMS purity: 100%, ¹ H NMR (400 MHz, DMSO-d6) δ 1.419-1.384 (m, 3H), 2.386 (s, 3H), 4.089 (s, 2H), 4.439-4.403 (m, 2H), 7.211 (s, 4H), 7.410 (s, 1H). Step -4. 3-(4-( bromomethyl ) benzyl ) iso Synthesis of oxadiazole -5- carboxylic acid ethyl ester (10_Int-5) .

Preparation of 3-(4-methylbenzyl)iso To a stirred solution of oxadiazole-5-carboxylic acid ethyl ester (10_Int-4) (1.8 g, 7.33 mmol, 1.0 eq) was added 1,2-dichloroethane (54 mL, 30 V), benzoyl peroxide (0.18 g, 0.73 mmol, 0.1 eq), and N-bromosuccinimide (1.3 g, 1.3 mmol, 1.0 eq) at RT. The RM was heated to 90 °C and stirred for 6 h. After completion of the reaction, the reaction mixture was quenched into _Na2S2O3 solution and extracted with DCM. The combined organic fractions were dried over _Na2SO4 ; concentrated and then purified by flash column chromatography (10-12% ethyl acetate in hexanes) to give compound (10_Int-5). MS (ES): 324.2 m/z [M]+ 326.2 [M+2]+, LCMS purity: 89.94%, ¹ H NMR (400 MHz, DMSO-d6) δ 1.286 (t, J=7.2 Hz, 3H), 4.070 (s, 2H), 4.325 (q, J=7.2 Hz, 2H), 4.685 (s, 2H), 7.155 (s, 1H), 7.290 (d, J=8 Hz, 2H), 7.401 (d, J=8 Hz, 2H). Step -5. 3-(4-( Cyanomethyl ) benzyl ) iso Synthesis of oxadiazole -5- carboxylic acid ethyl ester (10_Int-6) .

Preparation of 3-(4-(bromomethyl)benzyl)iso To a stirred solution of oxadiazole-5-carboxylic acid ethyl ester (10_Int-5) (0.3 g, 0.92 mmol, 1.0 eq) and acetone (6 mL, 20 V) was added tetrabutylammonium cyanide (1 g, 3.7 mmol, 4 eq) in nitrogen atmosphere and stirred at RT for 16 h. After the reaction was complete, the RM was quenched in water and extracted with ethyl acetate. The combined organic fractions were dried over Na ₂ SO ₄ ; concentrated and then purified by flash column chromatography (17-20% ethyl acetate in hexanes) to give compound (10_Int-6). MS (ES): 242.42 m/z [M-42]+, LCMS purity: 100%, ¹ H NMR (400 MHz, DMSO-d6) δ 1.287 (t, J=7.2 Hz, 3H), 4.004 (s, 2H), 4.072 (s, 2H), 4.327 (q, J=7.2 Hz, 2H), 7.133 (s, 1H), 3.343-7.291 (m, 4H).

Step-6. N-(4-carbamimidoylbenzyl)-3-(4-(cyanomethyl)benzyl)iso Oxazole-5-carboxamide (10) is prepared from 3-(4-(cyanomethyl)benzyl)iso Ethyl oxadiazole-5-carboxylate (10-Int-6) and 4-(aminomethyl)benzamidine dihydrochloride were prepared in a manner similar to that described in Example 2. MS (ES): 374.41 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 99.64% ¹ H NMR (400 MHz, DMSO-d6) δ 3.991 (s, 2H), 4.060 (s, 2H), 4.500 (s, 2H), 6.929 (s, 1H), 7.316 (s, 4H), 7.497 (d, J=7.2 Hz, 2H), 7.739 (d, J=7.2 Hz, 2H). Example 17 : Preparation of N-(4 -carbamimidoylbenzyl )-1-((4'- cyano- [1,1'- biphenyl ]-4- yl ) methyl )-1H- pyrazole -4- carboxamide (13) Step -1. 3-(4-( bromomethyl ) benzyl ) iso Synthesis of oxadiazole -5- carboxylic acid ethyl ester (13_Int-2) .

A stirred solution of 4'-methyl-[1,1'-biphenyl]-4-carbonitrile (13_Int-1) (0.1 g, 0.51 mmol, 1.0eq) and carbon tetrachloride (2 mL, 20V) was prepared. Then benzoyl peroxide (0.013 g, 0.05 mmol, 0.1eq) and N-bromosuccinimide (0.1 g, 0.56 mmol, 1.1eq) were added at RT. The RM was heated to 90 °C and stirred for ₁₆ h. After the reaction was complete, the RM was quenched into _Na2SO3 solution and extracted with DCM. The combined organic fractions were dried over _Na2SO4 ; concentrated and purified by flash column chromatography (2-5% ethyl acetate in hexanes) to give compound ( _{13_Int} -2). ¹ H NMR (400 MHz, DMSO-d6) δ 4.776 (s, 2H), 7.585 (d, J=8 Hz, 2H), 7.845 (d, J=8 Hz, 2H), 7.947 (s, 4H). Step -2. Synthesis of 1-((4'- cyano- [1,1'- biphenyl ]-4- yl ) methyl )-1H- pyrazole -4- carboxylic acid ethyl ester (13_Int-3)

Prepare a stirring solution of ethyl 1H-imidazole-4-carboxylate (0.25 g, 1.83 mmol, 1.0 eq) in acetone (10 mL, 20 V). Add 4'-(bromomethyl)-[1,1'-biphenyl]-4-carbonitrile (0.5 g, 1.89 mmol, 1.0 eq) and Cs ₂ CO ₃ (1.43 g, 1.40 mmol, 2.4 eq) at RT. Heat the RM to 65 °C and stir for 16 h. After completion of the reaction, cool the RM to RT, quench in water and extract with ethyl acetate. Dry the combined organic fractions over Na ₂ SO ₄ ; concentrate and purify by flash column chromatography (15-20% ethyl acetate in hexanes) to give compound (13_Int-3). MS (ES): 332.23 m/z [M+H]+.

Step-3. N-(4-Carbamimidoylbenzyl)-1-((4'-cyano-[1,1'-biphenyl]-4-yl)methyl)-1H-pyrazole-4-carboxamide (13) was prepared from ethyl 1-((4'-cyano-[1,1'-biphenyl]-4-yl)methyl)-1H-pyrazole-4-carboxylate (13_Int-3) and 4-(aminomethyl)benzamidine dihydrochloride in a similar manner to that described in Example-2. MS (ES): 435.37 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 99.83% ¹ H NMR (400 MHz, DMSO-d6) δ 4.485 (d, J=6 Hz, 2H), 5.424 (s, 2H), 7.340 (d, J=8 Hz, 2H), 7.495 (d, J=8 Hz, 2H), 7.758-7.739 (m, 4H), 7.858 (s, 1H), 7.879 (s, 1H), 7.940-7.920 (m, 3H), 8.329 (s, 1H), 8.794 (t, J=6 Hz, 1H), 8.984 (s, 1H), 9.248 (s, 2H). Example 18 : Preparation of N-(4- carbamimidoylbenzyl )-1-(4-(3- cyanobenzyl ) benzyl )-1H -pyrazole -4- carboxamide (28)

Step -1. N-(4-Carbamimidoylbenzyl)-1-(4-(3-cyanobenzyl)benzyl)-1H-pyrazole-4-carboxamide (28) was synthesized from ethyl 1-(4-(3-cyanobenzyl)benzyl)-1H-pyrazole-4-carboxylate (28_Int-4) and 4-(aminomethyl)benzamimidoyl dihydrochloride in a similar manner to that described in Example-2. MS (ES): 449.34.m/z [M+1]+, LCMS purity: 99.33%, HPLC purity: 100% ¹ H NMR (400 MHz, DMSO-d6) δ 3.98 (s, 2H), 4.47 (d, J = 6.40 Hz, 2H), 5.30 (s, 2H), 7.23 (q, J = 8.8 Hz, 4H), 7.50-7.47 (m, 2H), 7.56 (d, J = Hz, 1H), 7.66 (d, J = 7.60 Hz, 1H), 0.00 (d, J = 8.4 Hz, 2H), 7.89 (s, 1H), 8.25 (s, 1H), 8.75 (s, 2H), 8.90 (s, 2H), 9.24 (s, 2H). Example 19 : Preparation of N-(4- carbamimidoylbenzyl )-1-(4-(3- cyano -1H- pyrrol -1- yl ) benzyl )-1H- pyrazole -4 -carboxamide (25 ) Step -1. Synthesis of ethyl 1-(4- iodobenzyl )-1H- pyrazole -4- carboxylate (25_Int-2)

A stirred solution of ethyl 1H-pyrazole-4-carboxylate (1.3 g, 9.21 mmol, 1 eq) and 1-(bromomethyl)-4-iodobenzene (25_int-1) (3 g, 10.2 mmol, 1.1 eq) in acetone (13 mL, 10 V) was prepared and Cs ₂ CO ₃ (7.2 g, 0.20 mmol, 2.4 eq) was added at RT. The RM was heated to 65 °C and stirred for 8 h. After completion of the reaction, the RM was cooled to RT, quenched in water and extracted with ethyl acetate. The combined organic fractions were dried over Na ₂ SO ₄ ; concentrated and purified by flash column chromatography (30% ethyl acetate in hexanes) to give compound (28_Int-2). MS (ES): 357.27 m/z [M+1]+, LCMS purity: 82%. Step -2. Synthesis of ethyl 1-(4-(3- cyano -1H- pyrrol -1- yl ) benzyl )-1H- pyrazole -4- carboxylate (25_Int-3)

A stirred solution of ethyl 1-(4-iodobenzyl)-1H-pyrazole-4-carboxylate (28_Int-2) (0.800 g, 2.20 mmol, 1 eq) and 1H-pyrrole-3-carbonitrile (0.426 g, 8.95 mmol, 4 eq) in DMF (8 mL, 10 V) was prepared and K2CO3 (0.530 g, 6.78 mmol, 3 eq) and CuI (0.426 g, 2.23 mmol, 1 eq) were added at RT. The RM was heated to 100 °C and stirred for 8 h. After the reaction was completed, the RM was cooled at RT, quenched in water and extracted with ethyl acetate. The combined organic fractions were dried over _Na2SO4 , concentrated and purified by flash column chromatography (30% ethyl acetate in hexanes) to give compound (25_Int-3). _MS (ES): 319.04 m/z [M+1]+, LCMS purity: 70%.

Step -3. N-(4-Carbamimidoylbenzyl)-1-(4-(3-cyano-1H-pyrrol-1-yl)benzyl)-1H-pyrazole-4-carboxamide (25) was synthesized from ethyl 1-(4-(3-cyano-1H-pyrrol-1-yl)benzyl)-1H-pyrazole-4-carboxylate (25_Int-3) and 4-(aminomethyl)benzamimidoyl dihydrochloride in a similar manner to that described in Example-2. MS (ES): 424.15 m/z [M+1]+, LCMS purity: 100%, HPLC purity: 100% ¹ H NMR (400 MHz, DMSO- δ 4.49 (d, J = 5.20 Hz, 2H), 5.41 (s, 2H), 6.73 (s, 1H), 7.02 (s, 1H), 7.14 (s, 2H), 7.26 (s, 1H), 7.42 (d, J = 8.00 Hz, 1H), 0.00 (t, J = 10.8 Hz, 1H), 7.65 (d, J = 8.00 Hz, 1H), 7.76 (d, J = 7.60 Hz, 1H), 7.94 (s, 1H), 8.23 (s, δ -1H), 8.32 (s, 1H), 8.79 (s, 1H), 9.10 (s, 2H), 9.25 (s, 2H). ¹⁹ F NMR 400 MHz, DMSO-d6: δ -73.478 (1F). Example 20 : Preparation of N-(4- carbamimidoylbenzyl )-1-(4-(4- cyanobenzyl ) benzyl )-1H- pyrazole -4 - carboxamide (14) Step -1. Synthesis of 4-(4-( hydroxymethyl ) benzyl ) benzonitrile (14_Int-2)

Preparation of (4-cyanophenyl)boronic acid (1.5 g, 7.46 mmol, 1.0 eq) (14_Int-1) in 1,4-dihydro- To the stirred solution in 2-(bromomethyl)phenyl)methanol (1.1 g, 8.20 mmol, 1.1 eq), Cs ₂ CO ₃ (7.27 g, 22.38 mmol, 3 eq), PdCl 2 (dppf).DCM (3.04 g, 3.73 mol, 0.5 eq), and water were added at RT. The RM was heated to 100° C. and stirred for 8 h. After the reaction was completed, the RM was cooled to RT, quenched in water and extracted with ethyl acetate. The combined organic fractions were dried over Na ₂ SO ₄ ; concentrated and then purified by flash column chromatography (20% ethyl acetate in hexane) to give compound (14_Int-2). ¹ H NMR (400 MHz, DMSO-d6) δ 4.02 (s, 2H), 4.44 (s, 2H), 5.10 (t, J = 5.56 Hz, 1H), 7.22 (d, J = 8 Hz, 4H), 7.44 (d, J = 8.40 Hz, 2H), 7.74 (d, J = 8.40 Hz, 2H). Step -2. Synthesis of 4-(4-( chloromethyl ) benzyl ) benzonitrile (14-Int-3)

A stirring solution of ethyl 1H-pyrazole-4-carboxylate (0.300 g, 2.14 mmol, 1 eq) in DCM (10 mL, 10 V) was prepared and DIPEA (0.115 g, 8.96 mmol, 2 eq) and MsCl (0.107 g, 9.41 mmol, 2.1 eq) were added at 0 °C. The RM was stirred at RT for 6 h. After completion of the reaction, the RM was quenched in cold water and extracted with ethyl acetate. The combined organic fractions were dried over Na ₂ SO ₄ ; concentrated and then purified by flash column chromatography (4% ethyl acetate in hexanes) to give compound (14_Int-3). ¹ H NMR (400 MHz, DMSO-d6) δ 400 MHz, DMSO-d6: δ 4.05 (s, 2H), 4.72 (s, 2H), 7.22 (d, J = 8 Hz, 4H), 7.44 (d, J = 8.40 Hz, 2H), 7.74 (d, J = 8.40 Hz, 2H). Step -3. Synthesis of ethyl 1-(4-(4- cyanobenzyl ) benzyl )-1H- pyrazole -4 -carboxylate (25_Int-4)

A stirred solution of 1H-pyrazole-4-carboxylic acid ethyl ester (0.300 g, 2.14 mmol, 1 eq) and 4-(4-(chloromethyl)benzyl)benzonitrile (14_Int-3) (0.568 g, 2.35 mmol, 1.1 eq) in acetone (3 mL, 10 V) was prepared and Cs ₂ CO ₃ (1.67 g, 5.14 mmol, 2.4 eq) was added at RT. The RM was heated to 65 °C and stirred for 5 h. After completion of the reaction, the RM was cooled at RT, quenched in water and extracted with ethyl acetate. The combined organic fractions were dried over Na ₂ SO ₄ ; concentrated and purified by flash column chromatography (16% ethyl acetate in hexanes) to give compound (28_Int-2). MS (ES): 346.3 m/z [M+1]+, LCMS purity: 88%.

Step -4. N-(4-Carbamimidoylbenzyl)-1-(4-(4-cyanobenzyl)benzyl)-1H-pyrazole-4-carboxamide (14) was synthesized from ethyl 1-(4-(4-cyanobenzyl)benzyl)-1H-pyrazole-4-carboxylate (25_Int-4) and 4-(aminomethyl)benzamimidoyl dihydrochloride in a similar manner to that described in Example-2. MS (ES): 448 m/z [M+1]+, LCMS purity: 100%, HPLC purity: 100% ¹ H NMR (400 MHz, DMSO-d6) δ 4.02 (s, 2H), 4.47 (d, J = 6.00 Hz, 2H), 5.30 (s, 2H), 7.22 (s, 4H), 7.42 (d, J = 8.00 Hz, 2H), 7.49 (d, J = 8.40 Hz, 2H), 7.74 (d, J = 8.4 Hz, 3H), 0.00 (s, 1H), 8.25 (s, 1H), 8.76 (s, 1H), 8.92 (s, 2H), 8.95 (s, 2H). Example 21 : Preparation of N-(4- carbamimidoylbenzyl )-1-(4-(2- methoxyethyl ) benzyl )-1H- pyrazole -4- carboxamide (15) Step -1. Synthesis of ethyl 1-(4-(2- hydroxyethyl ) benzyl )-1H- pyrazole -4- carboxylate (15_Int-2)

A stirred solution of ethyl 1H-pyrazole-4-carboxylate (1 g, 7.14 mmol, 1.0 eq) (14_Int-1) in acetone (10 mL, 10 V) was prepared and 2-(4-(bromomethyl)phenyl)ethan-1-ol (1.6 g, 7.85 mmol, 1.1 eq) and Cs ₂ CO ₃ (5.58 g, 17.14 mmol, 2.4 eq) were added at RT. The RM was heated to 65 °C and stirred for 6 h. After completion of the reaction, the RM was cooled to RT, quenched in water and extracted with ethyl acetate. The combined organic fractions were dried over Na ₂ SO ₄ ; concentrated and purified by flash column chromatography (25% ethyl acetate in hexanes) to give compound (14_Int-2). MS (ES): 274.2 m/z [M+1]+, LCMS purity: 97%, HPLC purity: % ¹ H NMR (400 MHz, DMSO-d6) δ δ 1.22 (t, J = 7.20 Hz, 3H), 2.69 (t, J = 6.80 Hz, 2H), 3.57 (t, J = 6.80 Hz, 2H), 4.21 (t, J = 7.20 Hz, 2H), 4.63 (t, J = 5.20 Hz, 1H), 5.31 (s, 2H), 7.19 (s, 4H), 0.00 (s, 1H), 8.43 (s, 1H). Step -2. Synthesis of ethyl 1-(4-(2- methoxyethyl ) benzyl )-1H- pyrazole -4- carboxylate (15_Int-3)

A stirring solution of ethyl 1-(4-(2-hydroxyethyl)benzyl)-1H-pyrazole-4-carboxylate (15_Int-2) (0.900 g, 5.17 mmol, 1.0eq) in DMF (10 mL, 10V) was prepared and NaH (2.85 g, 10.34 mmol, 2eq) was added and stirred at 0 °C for 15 min. Then MeI (0.948 g, 6.724 mmol, 1.3eq) was added at 0 °C and the RM was stirred for 3 h. After completion of the reaction, the RM was quenched in cold water and extracted with ethyl acetate. The combined organic fractions were dried over Na ₂ SO ₄ ; concentrated and then purified by flash column chromatography (12% ethyl acetate in hexanes) to give compound (14_Int-3). MS (ES): 289.26 m/z [M+1]+, ¹ H NMR (400 MHz, DMSO-d6) δ 400 MHz, DMSO-d6: δ 1.26 (t, J = 6.80 Hz, 3H), 2.78 (t, J = 6.80 Hz, 2H), 3.27 (s, 3H), 3.51 (t, J = 6.80 Hz, 2H), 4.20 (q, J = 6.80 Hz, 2H), 5.31 (s, 2H), 7.20 (s, 2H), 0.00 (s, 2H), 8.43 (s, 2H).

Step -3. N-(4-Carbamimidoylbenzyl)-1-(4-(2-methoxyethyl)benzyl)-1H-pyrazole-4-carboxamide (15) was synthesized from ethyl 1-(4-(2-methoxyethyl)benzyl)-1H-pyrazole-4-carboxylate (15_Int-3) and 4-(aminomethyl)benzamimidoyl dihydrochloride in a similar manner to that described in Example-2. MS (ES): 392.7 m/z [M+1]+, LCMS purity: 100%, HPLC purity: 100% ¹ H NMR (400 MHz, DMSO-d6) δ 2.52 (t, J = 181.60 Hz, 2H), 3.21 (s, 3H), 3.50 (t, J = 6.8 Hz, 2H), 4.48 (d, J = 5.60 Hz, 2H), 5.30 (s, 2H), 7.20 (t, J = 8.40 Hz, 4H), 7.49 (d, J = 8.40 Hz, 2H), 0.00 (d, J = 8.00 Hz, 2H), 7.90 (s, 1H), 8.26 (s, 1H), 8.77 (s, 1H), 8.97 (s, 2H), 9.25 (s, 2H). Example 22 : Preparation of N-(4- carbamimidoyl -3- fluorobenzyl )-1-(4-(3- cyanobenzyl ) benzyl )-1H -pyrazole -4 -carboxamide (49)

Step 1: Preparation of N-(4-carbamimidoyl-3-fluorobenzyl)-1-(4-(3-cyanobenzyl`1)benzyl)-1H-pyrazole-4-carboxamide (49) was prepared from 1-(4-(3-cyanobenzyl)benzyl)-1H-pyrazole-4-carboxylic acid ethyl ester (28-Int-4) and 4-(aminomethyl)-2-fluorobenzamidine dihydrochloride (43-Int-5) in a similar manner to that described in Example-2. MS (ES): 467.14 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 100% ¹ H NMR 400 MHz, DMSO-d6: δ 3.99 (s, 2H), 4.46 (d, J = 6.00 Hz, 2H), 5.34 (s, 2H), 7.34-7.20 (m, 6H), 7.48 (t, J = 7.60 Hz, 1H), 7.71-7.56 (m, 3H), 7.89 (s, 1H), 8.25 (s, 1H), 8.76 (t, J = 6 Hz, 1H), 9.18 (s, 2H), 9.36 (s, 2H), ¹⁹ F NMR 400 MHz, DMSO-d6: δ -73.492 (1F), -113.893 (0.35F). Example 23 : N-(4- Carbamimidoyl -2- fluorobenzyl )-1-(4-(3- cyanobenzyl ) benzyl )-1H- pyrazole -4- carboxamide (50)

N-(4-Carbamimidoyl-2-fluorobenzyl)-1-(4-(3-cyanobenzyl)benzyl)-1H-pyrazole-4-carboxamide (50) was prepared from ethyl 1-(4-(3-cyanobenzyl)benzyl)-1H-pyrazole-4-carboxylate (28-Int-4) and 4-(aminomethyl)-3-fluorobenzamidine dihydrochloride (45-Int5) in a similar manner as described in Example-2. MS (ES): 466.52 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 100% ¹ H NMR 400 MHz, DMSO-d6: δ 3.99 (s, 2H), 3.32 (s, 1H), 4.49 (d, J = 5.60 Hz, 1H), 5.30 (s, 1H), 7.23 (q, J = 8.00 Hz, 5H), 7.71-7.48 (m, 8H), 7.90 (s, 1H), 8.25 (s, 1H), 8.73 (s, 1H), 9.11 (s, 1H), 9.32 (s, 1H), ¹⁹ F NMR 400 MHz, DMSO-d6: δ -73.536 (1F), -117.182 (0.68F). Example 24 : Preparation of N-(4 -carbamimidoylbenzyl )-1-(3-(2- methoxyethyl ) benzyl )-1H- pyrazole -4 -carboxamide (69) Step 1. Synthesis of 1- bromo -3-(2- methoxyethyl ) benzene (69_Int-2)

A stirred solution of 2-(3-bromophenyl)ethan-1-ol (69_Int-1) (3 g, 14.92 mmol, 1.0 eq) in DMF (30 mL, 10 V) was prepared and NaH (60% in mineral oil) (0.72 g, 17.91 mmol, 1.2 eq) was slowly added at 0 °C and stirred for 1 h. Iodomethane (2.54 g, 17.91 mmol, 1.2 eq) was added to the RM at 0 °C and then brought to RT and stirred for 16 h. After the reaction was complete, the RM was slowly quenched with cold water and extracted with ethyl acetate. The combined organic fractions were dried _{over Na2SO4} ; concentrated and purified by flash column chromatography (4-5% ethyl acetate in hexanes) to give compound (69_Int-2) MS (ES): 162.23 m/z [M-53]+, LCMS purity: 99.25%, 1H NMR (400 MHz, DMSO-d6) δ 2.808 (t, J=6.4 Hz, 2H), 3.235 (s, 3H), 3.533 (t, J=6.4 Hz, 2H), 7.250 (d, J=5.2 Hz, 2H), 7.383-7.410 (m, 1H), 7.458 (s, 1H). Step 2. Synthesis of 3-(2- methoxyethyl ) benzaldehyde (69_Int-3)

Prepare a stirred solution of 1-bromo-3-(2-methoxyethyl)benzene (69_Int-2) (1.5 g, 7.0 mmol, 1.0 eq) in diethyl ether (15 mL, 10 V) and add tetramethylethylenediamine (1.75 g, 1.5 mmol, 2.16 eq) at RT. Cool the RM to -75 °C and then add n-BuLi (2.5 M in hexane) (5.6 mL, 14.0 mmol, 2.0 eq) dropwise under _N2 . Stir the RM at -75 °C for 1 h. Then warm the RM to -20 °C and stir for an additional 20 min. Cool the RM again to -75 °C and then add anhydrous DMF (7.5 mL, 5 V) dropwise and stir for 16 h. After completion of the reaction, the RM was slowly quenched with 1N HCl and extracted with ethyl acetate. The combined organic fractions were dried _{over Na2SO4} ; concentrated and purified by flash column chromatography (10-12% ethyl acetate in hexanes) to give compound (69_Int-3) MS (ES): 132.8 m/z [M-32]+, LCMS purity: 100%, 1H NMR (400 MHz, DMSO-d6) δ 2.902 (t, J=6.4 Hz, 2H), 3.325 (s, 3H), 3.581 (t, J=6.4 Hz, 2H), 7.518 (t, J=7.6 Hz, 1H), 7.588 (d, J=8.0 Hz, 1H), 7.742-7.767 (m, 2H), 9.983 (s, 1H). Step 3. Synthesis of (3-(2- methoxyethyl ) phenyl ) methanol (69_Int-4)

A stirring solution of 3-(2-methoxyethyl)benzaldehyde (69_Int-3) (0.23 g, 1.40 mmol, 1.0 eq) in methanol (4.6 mL, 20 V) was prepared and sodium borohydride (0.106 g, 2.8 mmol, 2.0 eq) was slowly added at RT and stirred for 4 h. After the reaction was complete, the RM was evaporated. The residue was quenched with water, adjusted to pH ~5 with 2N HCl, and extracted with ethyl acetate. The combined organic fractions were dried _{over Na2SO4} ; concentrated and purified by flash column chromatography (12-15% ethyl acetate in hexanes) to give compound (69_Int-4) MS (ES): 149.1 m/z [M-18]+, LCMS purity: 97.53%, 1H NMR (400 MHz, DMSO-d6) δ 2.794 (t, J=6.8 Hz, 2H), 3.240 (s, 3H), 3.527 (t, J=6.8 Hz, 2H), 4.467 (d, J=5.6 Hz, 2H), 5.150 (t, J=6.0 Hz, 1H), 7.092 (d, J=7.6 Hz, 1H), 7.151 (t, J = 7.6 Hz, 2H), 7.231 (t, J = 7.6 Hz, 1H). Step 4. Synthesis of 1-( bromomethyl )-3-(2- methoxyethyl ) benzene (69_Int-5)

A stirred solution of (3-(2-methoxyethyl)phenyl)methanol (69_Int-4) (0.2 g, 1.20 mmol, 1.0 eq) in DCM (4 mL, 20 V) was prepared. Then triphenylphosphine (0.34 g, 1.32 mmol, 1.1 eq) and carbon tetrabromide (0.44 g, 1.32 mmol, 1.1 eq) were added at RT and stirred for 16 h. After the reaction was completed, the RM was quenched in water and extracted with DCM. The combined organic fractions were dried _{over Na2SO4} ; concentrated and purified by flash column chromatography (8-10% ethyl acetate in hexanes) to give compound (69_Int-5) MS (ES): 248.2 m/z [M+18]+, LCMS purity: 100%, 1H NMR (400 MHz, DMSO-d6) δ 2.816 (t, J=6.8 Hz, 2H), 3.343 (s, 3H), 3.538 (t, J=6.8 Hz, 2H), 4.681 (s, 2H), 7.187 (s, 1H), 7.259-7.309 (m, 2H), 7.332-7.387 (m, 1H). Step 5. Synthesis of ethyl 1-(3-(2- methoxyethyl ) benzyl )-1H- pyrazole -4- carboxylate (69_Int-6)

Prepare a stirred solution of 1-(bromomethyl)-3-(2-methoxyethyl)benzene (69_Int-5) (0.176 g, 0.76 mmol, 1.0 eq) in acetone (3.52 mL, 20 V). Then add ethyl 1H-pyrazole-4-carboxylate (0.4 g, 0.76 mmol, 1.0 eq) and cesium carbonate (0.6 g, 1.84 mmol, 2.4 eq) at RT. Bring the RM to 65 °C and stir for 16 h. After the reaction was completed, the RM was filtered and the filtrate was concentrated and then purified by flash column chromatography (20-25% ethyl acetate in hexane) to give compound (69_Int-6) MS (ES): 289.4 m/z [M+H]+, LCMS purity: 100%, 1H NMR (400 MHz, DMSO-d6) δ 1.25 (t, J=8.0 Hz, 3H), 2.77 (t, J=8.0 Hz, 2H), 3.22 (s, 3H), 3.50 (d, J=8.0 Hz, 2H), 4.12 (q, J=8.0 Hz, 2H), 5.33 (s, 2H), 7.08 (d, J=8.0 Hz, 1H), 7.15-7.18 (m, 2H), 7.25 (t, J=8.0 Hz, 1H), 7.87 (s, 1H), 8.45 (s, 1H). Step 6. Synthesis of N-(4- carbamimidoylbenzyl )-1-(3-(2- methoxyethyl ) benzyl )-1H- pyrazole -4- carboxamide (69)

A stirred solution of ethyl 1-(3-(2-methoxyethyl)benzyl)-1H-pyrazole-4-carboxylate (69_Int-6) (0.1 g, 0.34 mmol, 1.0 eq) in THF (1 mL, 10 V) was prepared. 4-(Aminomethyl)benzamididine dihydrochloride (0.09 g, 0.41 mmol, 1.2 eq) and DIPEA (0.18 g, 1.38 mmol, 4.0 eq) were added at RT. The RM was then cooled to 0 °C and TMA (2M in toluene) (1.0 mL, 2.08 mmol, 6.0 eq) was added. The RM was brought to 85 °C and stirred for 16 h. After the reaction was complete, the RM was quenched by 1 V of water and evaporated. The residue was washed with 20% methanol in dichloromethane and filtered. The filtrate was concentrated and purified by PREP HPLC ((A) 0.1% TFA in water (B) 100% MeCN). Finally, the pure fractions were lyophilized to give compound (69) MS (ES): 392.6 m/z [M+H]+, LCMS purity: 97.92%, HPLC purity: 100% 1H NMR (400 MHz, DMSO-d6) δ 2.790 (t, J=6.8 Hz,, 2H), 3.226 (s, 3H), 3.522 (d, J=6.8 Hz, 2H), 4.492 (d, J=6.4 Hz, 2H), 5.326 (s, 2H), 7.088 (d, J=8.0 Hz, 1H), 7.077-7.131 (m, 2H), 7.181 (t, J=4.4 Hz, 1H), 7.403 (d, 3H), 7.761 (d, J=8.4 Hz, 2H), 7.924 (s, 1H), 8.274 (s, 1H), 8.776 (t, J=6.0 Hz, 1H), 9.043 (s, 2H), 9.256 (s, 2H). Example 25 : Preparation of N-(4- carbamimidoyl -3,5 -difluorobenzyl )-1-(4-(2- cyanopropan -2- yl ) benzyl )-1H- pyrazole -4- carboxamide (44) Step -1. Synthesis of N-(4- carbamimidoyl -3,5- difluorobenzyl )-1-(4-(2- cyanoprop -2- yl ) benzyl )-1H- pyrazole -4- carboxamide (44)

Prepare a stirred solution of ethyl 1-(4-(2-cyanopropan-2-yl)benzyl)-1H-pyrazole-4-carboxylate (7_Int-5) (0.1 g, 0.3367 mmol, 1.0eq) in toluene (2 mL, 10V). 4-(Aminomethyl)-2,6-difluorobenzamidine dihydrochloride (44_Int-8) (0.095 g, 0..505 mmol, 1.5eq) and DIPEA (0.6 mL, 0.8417 mmol, 2.5eq) are added to the RM at RT. The RM is cooled to 0 °C and TMA (2.0M in toluene, 0.6 mL, 1.0101 mmol, 3.0eq) is added. The RM is then heated to 100 °C and stirred for 16 h. After completion of the reaction, the RM was quenched with 1V of water and evaporated. The residue was washed with 20% methanol in dichloromethane and filtered. The solid residue was discarded and the filtrate was concentrated and then purified by PREP HPLC ((A) 0.1% TFA in water (B) 100% MeCN). Finally, the pure fractions were lyophilized to give compound (44). MS (ES): 437.21 m/z [M+1]+, LCMS purity: 100%, HPLC purity: 94.56% 1H 400 MHz, DMSO-d6: δ 1.664 (s, 6H), 4.462 (d, J = 6.40 Hz, 2H), 5.369 (s, 2H), 7.252 (d, J=8.8 Hz, 2H), 7.331 (d, J = 8.0 Hz, 2H), 7.511 (d, J = 8.00 Hz, 2H), 7.921 (s, 1H), 8.310 (s, 1H), 8.840 (t, J=6.4 Hz, 1H), 9.513 (s, 2H), 9.669 (s, 2H). Example 26 : N-(4- Carbamimidoylbenzyl )-1-(4-(2- cyanoprop -2 -yl ) benzyl )-5- cyclopropyl -1H- pyrazole -4- carboxamide (83) Step -1. Synthesis of 2-(4-(2- cyanopropan -2- yl ) benzyl ) hydrazine -1- carboxylic acid tributyl ester (83_Int-1)

A stirring solution of 2-(4-(bromomethyl)phenyl)-2-methylpropionitrile (7_Int-3) (0.5 g, 2.1 mmol, 1.0 eq) in dimethylacetamide (5 mL, 10V) was prepared. DIPEA (0.54 g, 4.2 mmol, 2.0 eq) and tributyl hydrazinecarboxylate (0.55 g, 4.2 mmol, 2.0 eq) were added at RT and stirred at 70 °C for 16 h. After the reaction was complete, the RM was quenched in water and extracted with ethyl acetate. The combined organic fractions were washed with saline solution, dried over Na2SO4, concentrated, and purified by flash column chromatography (10-12% ethyl acetate in hexane) to give compound (83_Int-1), 1H 400 MHz, DMSO-d6: δ 1.637 (s, 6H), 1.760 (s, 9H), 4.206 (s, 2H), 7.514 (s, 4H), 7.744 (d, br, J = 8.00 Hz, 1H), 7.886-8.004 (d, br, J = 47.2, 1H). Step -2. Synthesis of 2-(4-( hydrazinomethyl ) phenyl )-2- methylpropionitrile (83_Int-2)

A stirred solution of tributyl 2-(4-(2-cyanopropan-2-yl)benzyl)hydrazine-1-carboxylate (83_Int-1) (2.0 g, 6.9 mmol, 1.0 eq) in methanol (10 mL, 5 V) was prepared and water (22 mL, 11 V) and concentrated HCl (6.6 mL, 3.3 V) were added at RT. The RM was heated to 80 °C and stirred for 16 h. After the reaction was complete, the RM was evaporated and triturated in ethyl acetate to give compound (83_Int-2). MS (ES): 191.4 m/z [M+H]+, LCMS purity: 83.16%, 1H 400 MHz, DMSO-d6: δ 1.693 (s, 6H), 4.076 (s, 2H), 7.096 (s, br, 2H), 7.517 (d, J = 12.8 Hz, 4H). Step -3. Synthesis of 1-(4-(2- cyanopropan -2- yl ) benzyl )-5 - cyclopropyl -1H- pyrazole -4- carboxylic acid ethyl ester (7-Int-3)

A solution of 3-cyclopropyl-3-oxopropanoate (0.22 g, 1.41 mmol, 1.0 eq) and dimethylformamide dimethylacetal (0.185 g, 1.55 mmol, 1.1 eq) was prepared and stirred at 75 °C for 90 min. The RM was cooled to RT and then ethanol (4.4 mL, 20 V), TEA (0.57 g, 5.63 mmol, 4.0 eq), and 2-(4-(hydrazinomethyl)phenyl)-2-methylpropionitrile (83_Int-2) (0.365 g, 1.41 mmol, 1.0 eq) were added. The RM was heated to 80 °C and stirred for 16 h. After the reaction was complete, the RM was quenched into water and extracted with ethyl acetate. The combined organic fractions were washed with saline solution, dried over Na2SO4, concentrated, and purified by flash column chromatography (12-15% ethyl acetate in hexanes) to give compound (83_Int-3). MS (ES): 180.88 m/z [M+H]+, LCMS purity: 89%, 1H 400 MHz, DMSO-d6: δ 1.637 (s, 6H), 4.174-4.225 (m, 2H), 5.484 (s, 2H), 7.222 (d, J = 8.0 Hz, 2H), 7.505 (d, J=8.0 Hz, 2H), 7.830 (s, 1H). Step -4. Synthesis of N-(4- carbamimidoylbenzyl )-1-(4-(2- cyanoprop -2- yl ) benzyl )-5- cyclopropyl -1H- pyrazole -4- carboxamide (83)

The final compound was prepared from ethyl 1-(4-(2-cyanopropan-2-yl)benzyl)-5-cyclopropyl-1H-pyrazole-4-carboxylate (7-Int-3) and 4-(aminomethyl)benzamididine dihydrochloride in a manner similar to that described in (Example 24) (83). MS (ES): 441.4 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 100% 1H 400 MHz, DMSO-d6: δ 0.734-0.757 (m, 2H), 0.919-0.967 (m, 2H), 1.658 (s, 6H), 1.753-1.796 (m, 1H), 4.472 (d, J=6.0 Hz, 2H), 5.438 (s, 2H), 7.227 (d, J = 8 Hz, 2H), 7.501 (t, J=8.8 Hz, 4H), 7.762 (d, J=8.0 Hz, 2H), 7.801 (s, 1H), 8.534 (t, J = 6.0 Hz, 1H), 8.977 (s, 2H), 9.258 (s, 2H). Example 27 : N-(4- Carboxamidinobenzyl )-1-(4-(2- cyanoprop -2- yl ) benzyl )-5- propyl -1H- imidazole -4- carboxamide (84) Step -1. Synthesis of ethyl 1-(4-(2- cyanopropan -2- yl ) benzyl )-5- propyl -1H- imidazole -4- carboxylate (84_Int-1)

A stirred solution of ethyl 5-propyl-1H-imidazole-4-carboxylate (0.45 g, 2.47 mmol, 1.0 eq) in DMF (4.5 mL, 10 V) was prepared and NaHMDS (1M in THF, 2.5 mL, 2.47 mmol, 1.0 eq) was added at 0 °C and stirred for 30 min. 2-(4-(Bromomethyl)phenyl)-2-methylpropionitrile (7_Int-3) (0.5 g, 3.56 mmol, 1.0 eq) was added to the RM at 0 °C, brought to RT, and stirred for 16 h. After the reaction was complete, the RM was quenched into water and extracted with ethyl acetate. The combined organic fractions were dried over Na2SO4; concentrated and then purified by flash column chromatography (20-25% ethyl acetate in hexanes). (84_Int-1). Confirmed by 2D NMR (ROE analysis). MS (ES): 339.9 m/z [M+H]+, LCMS purity: 84.72%, 1H 400 MHz, DMSO-d6: δ 0.807 (t, J = 7.2 Hz, 3H), 1.251 (q, J = 8.0 Hz, 3H), 1.297 (s, 2H), 1.654 (s, 6H), 2.734 (q, J = 6.4 Hz, 2H), 4.186 (q, J=7.2 Hz, 2H), 5.264 (s, 2H), 7.194 (d, J=8.0 Hz, 2H), 7.512 (d, J=8.0 Hz, 2H), 7.788 (s, 1H). Step -2. Synthesis of N-(4- carbamimidoylbenzyl )-1-(4-(2- cyanoprop -2- yl ) benzyl )-5- propyl -1H- imidazole -4- carboxamide (84)

The final compound was prepared from ethyl 1-(4-(2-cyanopropan-2-yl)benzyl)-5-propyl-1H-imidazole-4-carboxylate (84-Int-1) and 4-(aminomethyl)benzamidinium dihydrochloride in a manner similar to that described in (Example-24) (84). MS (ES): 443.35 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 99.63% 1H 400 MHz, DMSO-d6: δ 0.782 (t, J = 7.2 Hz, 3H), 1.262-1302 (m, 2H), 1.656 (d, J = 6.00 Hz, 2H), 2.773 (t, J = 8 Hz, 2H), 4.473 (d, J=6.4 Hz, 2H), 5.260 (s, 2H), 7.226 (d, J = 8.4 Hz, 2H), 7.487-7.526 (m, 4H), 7.745 (d, J=8.4 Hz, 2H), 7.872 (s, 1H), 8.634 (t, J=6.0 Hz, 1H), 9.069 (s, 2H), 9.248 (s, 2H). Example 28 : Preparation of N-(4- carbamimidoylbenzyl )-1-(4-(2- cyanoprop -2- yl ) benzyl )-5- methyl -1H- pyrazole -4- carboxamide (81) Step -1. Synthesis of 5- methyl -1H- pyrazole -4- carboxylic acid ethyl ester (81_Int-2)

A solution of ethyl 3-oxobutyrate (81_Int-1) (0.5, 3.84 mmol, 1.0 eq) and dimethylformamide dimethylacetal (0.46 g, 3.84 mmol, 1.0 eq) was prepared and stirred at 110 °C for 1 h. The RM was cooled to RT followed by the addition of ethanol (5.0 mL, 10 V) and hydrazine hydrate (99%) (0.2 g, 3.84 mmol, 1.0 eq). The RM was heated to 70 °C and stirred for 2 h. After completion of the reaction, the reaction mixture was evaporated to give compound (81_Int-2). MS (ES): 154.96 m/z [M+H]+, LCMS purity: 100%, 1H NMR (400 MHz, DMSO-d6) δ 1.241 (t, J=8.00 Hz, 3H), 2.446 (s, 3H), 4.178 (q, J=4.2 Hz, 2H), 7.794 (s, 1H), 13.102 (s, 1H). Step -2. Synthesis of 1-(4-(2- cyanopropan -2- yl ) benzyl )-5- methyl -1H- pyrazole -4- carboxylic acid ethyl ester (81_Int-3)

A stirred solution of ethyl 5-methyl-1H-pyrazole-4-carboxylate (81_Int-2) (0.25 g, 1.62 mmol, 1.0 eq) in DMF (2.5 mL, 10 V) was prepared and NaHMDS (1M in THF) (1.62 mL, 1.62 mmol, 1.0 eq) was added at 0 °C and stirred for 30 min. 2-(4-(Bromomethyl)phenyl)-2-methylpropionitrile (7_Int-3) (0.4 g, 1.62 mmol, 1.0 eq) was added to the RM at 0 °C. The RM was brought to RT and stirred for 16 h. After the reaction was complete, the RM was quenched into water and extracted with ethyl acetate. The combined organic fractions were dried over Na2SO4; concentrated and purified by flash column chromatography (20-25% ethyl acetate in hexanes) to give compound (81_Int-3). MS (ES): 312.2 m/z [M+H]+, LCMS purity: 100%, 1H 400 MHz, DMSO-d6: δ 1.239 (t, J = 8.00 Hz, 3H), 1.656 (s, 6H), 2.299 (s, 3H), 4.171 (t, J = 8.00 Hz, 2H), 5.273 (s, 2H), 7.190 (d, J=8.00 Hz, 2H), 7.322 (d, J=8.00 Hz, 2H), 7.829 (s, 1H). Step -3. Synthesis of N-(4- carbamimidoylbenzyl )-1-(4-(2- cyanoprop -2- yl ) benzyl )-5- propyl -1H- imidazole -4- carboxamide (81)

The final compound was prepared from ethyl 1-(4-(2-cyanopropan-2-yl)benzyl)-5-propyl-1H-imidazole-4-carboxylate (81-Int-3) and 4-(aminomethyl)benzamididine dihydrochloride in a manner similar to that described in (Example 25) (81). MS (ES): 415.35 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 99.47% 1H 400 MHz, DMSO-d6: δ 1.655 (s, 6H), 2.457 (s, 3H), 4.777 (d, J = 6.0 Hz, 2H), 5.341 (s, 2H), 7.191 (d, J=8 Hz, 2H), 7.515-7.477 (m, 4H), 7.750 (d, J = 8 Hz, 2H), 7.985 (s, 1H), 8.676(t, J=6 Hz, 1H), 8.886 (s, 2H), 9.247 (s, 2H). Example 29 : N-(4- Carbamimidoyl -3- fluoro -5- methoxybenzyl )-1-(4-(2- cyanopropan -2- yl ) benzyl )-1H- pyrazole -4- carboxamide (48) Step -1. Synthesis of tributyl (4- cyano -3 - fluoro -5- methoxybenzyl ) carbamate (48_Int-1)

Prepare a stirring solution of tributyl (4-cyano-3,5-difluorobenzyl)carbamate (44_Int-7) (5 g, 18.36 mmol, 1.0 eq) in THF (50 mL, 10 V) and MeOH (50 mL, 10 V). Cool the RM to 0 °C and add sodium methoxide (4.02 g, 74.6 mmol, 4.0 eq) slowly. Bring the RM to RT and stir for 5 h. After the reaction is complete, quench the RM into cold water and extract with ethyl acetate. The combined organic fractions were dried over Na2SO4 and concentrated to give compound (48_Int-1) 1H 400 MHz, DMSO-d6: δ 1.631 (s, 9H), 3.924 (s, 3H), 4.169 (d, J = 6.0 Hz, 2H), 6.871 (d, J = 10 Hz, 1H), 6.982 (s, 1H), 7.544 (t, J = 6.0 Hz, 1H). Step -2. Synthesis of tributyl (3- fluoro -4-(N- hydroxycarbamimidoyl )-5- methoxybenzyl ) carbamate (48_Int-2)

A stirred solution of tributyl (4-cyano-3-fluoro-5-methoxybenzyl)carbamate (48_Int-1) (4.7 g, 16.78 mmol, 1.0 eq) in MeOH (50 mL, 10V) was prepared. Then hydroxylamine hydrochloride (1.96 gm, 28.536 mmol, 1.7 eq) and DIPEA (5.0 mL, 28.535 mmol, 1.7 eq) were added at RT. The RM was heated to 70 °C and stirred for 16 h. After completion of the reaction, the RM was cooled to RT, quenched into water, and extracted with DCM. The combined organic fractions were dried over Na2SO4; concentrated to give compound (48_Int-2). MS (ES): 314.29 m/z [M+1]+, LCMS purity: 80%. Step -3. Synthesis of tributyl (4- carbamimidoyl -3- fluoro -5- methoxybenzyl ) carbamate (48_Int-3)

Prepare a stirred solution of tributyl (3-fluoro-4-(N-hydroxycarbamimidoyl)-5-methoxybenzyl)carbamate (48_Int-2) (2 g, 6.3397 mmol, 1.0 eq) in MeOH. Then add (20 mL, 10V), ammonium chloride (1.7 g, 31.9485 mmol, 5.0eq), and iron (1.7 g, 31.9485 mmol, 5.0eq) at RT. Cool the RM to 0 °C and then add acetic acid (20 mL, 5V) dropwise. Heat the RM to 70 °C and stir for 16 h. After the reaction is complete, cool the RM to RT. The RM was concentrated, quenched in cold water and basified to target pH 10 by slow addition of saturated NaOH solution. The solid was filtered and the filtrate was extracted with DCM. The combined organic fractions were dried over Na2SO4; concentrated to give compound (48_Int-3). MS (ES): 298.23 m/z [M+1]+, LCMS purity: 73.2%. Step -4. Synthesis of 4-( aminomethyl )-2- fluoro -6- methoxybenzamidine (48_Int-4) .

A stirring solution of (4-carbamimidoyl-3-fluoro-5-methoxybenzyl)carbamic acid tributyl ester (48_Int-3) (0.8 g, 25.473 mmol, 1.0 eq) in water (8 mL, 10 V) was prepared and concentrated HCl (2.6 mL, 3.3 V) was added at RT and stirred for 3 h. After completion of the reaction, the RM was concentrated and triturated in methanol to give compound (43_Int-4). MS (ES): 198.2 m/z [M+1]+, LCMS purity: 36%. Step -5. Synthesis of N-(4- carbamimidoyl -3 - fluoro -5- methoxybenzyl )-1-(4-(2- cyanopropan -2- yl ) benzyl )-1H- pyrazole -4- carboxamide (48) .

The final compound was prepared from ethyl 1-(4-(2-cyanopropan-2-yl)benzyl)-1H-pyrazole-4-carboxylate (7-Int-5) and 4-(aminomethyl)-2-fluoro-6-methoxybenzamidine dihydrochloride (48-Int-4) in a similar manner as described in (Example-25) (48). MS (ES): 449.22 m/z [M+1]+, LCMS purity: 93.36%, HPLC purity: 93.46% 1H 400 MHz, DMSO-d6: δ 1.660 (s, 6H), 3.843 (s, 3H), 4.436 (d, J = 5.60 Hz, 2H), 5.364 (s, 2H), 6.878 (d, J=10 Hz, 2H), 6.966 (s, 1H), 7.324 (d, J = 8 Hz, 2H), 7.507 (d, J=8.0 Hz, 2H), 7.919 (s, 1H), 8.299 (s, 1H), 8.778 (t, J= 6.4 Hz, 1H), 9.139 (s, 2H), 9.383 (s, 2H). Example 30 : N-(4- Carbamimidoyl -3- fluorobenzyl )-1-(4-( cyanomethyl ) benzyl )-1H- pyrazole -4 -carboxamide (72) Step -1. Synthesis of N-(4- carbamimidoyl -3- fluorobenzyl )-1-(4-( cyanomethyl ) benzyl )-1H- pyrazole -4- carboxamide (72)

The final compound was prepared from ethyl 1-(4-(cyanomethyl)benzyl)-1H-pyrazole-4-carboxylate (1-Int-4) and 4-(aminomethyl)-2-fluorobenzamidine dihydrochloride (43-Int-5) in a manner similar to that described in (Example-25) (72). MS (ES): 391.22 m/z [M+1]+, LCMS purity: 100%, HPLC purity: 100% 1H 400 MHz, DMSO-d6: δ 4.024 (s, 2H), 4.460 (s, 2H), 5.357 (s, 2H), 7.304-3.349 (m, 6H), 7.622 (s, 1H), 7.915 (s, 1H), 8.280 (s, 1H), 8.800 (t, J=6.4 Hz, 1H), 9.132 (s, 2H), 9.377 (s, 2H). Example 31 : N-(4- carbamimidoyl -3- fluorobenzyl )-1-(4-(2- methoxyethyl ) benzyl )-1H- pyrazole -4- carboxamide (73) Step -1. Synthesis of N-(4- carbamimidoyl -3- fluorobenzyl )-1-(4-(2- methoxyethyl ) benzyl )-1H- pyrazole -4- carboxamide (73)

The final compound was prepared from ethyl 1-(4-(2-methoxyethyl)benzyl)-1H-pyrazole-4-carboxylate (15-Int-3) and 4-(aminomethyl)-2-fluorobenzamidine dihydrochloride (43-Int-5) in a manner similar to that described in (Example-25) (73). MS (ES): 410.4 m/z [M+1]+, LCMS purity: 100%, HPLC purity: 99.67% 1H 400 MHz, DMSO-d6: δ 2.774 (t, J =6.80 Hz, 2H), 3.212 (s, 3H), 3.500 (t, J =6.80 Hz, 2H), 7.460 (d, J=6.0 Hz, 2H), 5.303 (s, 2H), 7.175-7.224 (m, 4H), 7.319 (t, J = 7.2 Hz, 2H), 7.899 (s, 1H), 8.260 (s, 1H), 8.797 (t, J=6.0 Hz, 1H), 9.182 (s, 2H), 9.377 (s, 2H). Example 32 : N-(4- Carbamimidoyl- 3- fluorobenzyl )-1-(4-( cyanomethyl ) benzyl )-5-( methoxymethyl )-1H- pyrazole -4- carboxamide (75) Step -1. Synthesis of N-(4- carbamimidoyl -3- fluorobenzyl )-1-(4-( cyanomethyl ) benzyl )-5-( methoxymethyl )-1H- pyrazole -4- carboxamide (75)

The final compound was prepared from methyl 1-(4-(cyanomethyl)benzyl)-5-(methoxymethyl)-1H-pyrazole-4-carboxylate (85-Int-2) and 4-(aminomethyl)-2-fluorobenzamidine dihydrochloride (43-Int-5) in a similar manner as that described in (Example-24) (75). MS (ES): 435.4 m/z [M+1]+, LCMS purity: 100%, HPLC purity: 100% 1H 400 MHz, DMSO-d6: δ 3.232 (s, 3H), 4.019 (s, 2H), 4.85 (d, J = 6.0 Hz, 2H), 4.812 (s, 2H), 5.365 (s, 2H), 7.242 (d, J = 7.6 Hz, 2H), 7.307-7.374 (M, 4H), 7.640 (t, = Hz, 1H), 8.039 (s, 1H), 8.871 (t, J=6.0 Hz, 1H), 9.197 (s, 2H), 9.401 (s, 2H). Example 33 : N-(4- carbamimidoylbenzyl )-1-(4-( methoxymethyl ) benzyl )-1H- pyrazole -4- carboxamide (71) Step -1. Synthesis of ethyl 1-(4-( methoxymethyl ) benzyl )-1H- pyrazole -4- carboxylate (71_Int-1)

A stirred solution of ethyl 1-(4-(hydroxymethyl)benzyl)-1H-pyrazole-4-carboxylate (36_Int-2) (2.5 g, 9.6153 mmol, 1.0 eq) in DMF (25 mL, 10 V) was prepared and NaH (60% in mineral oil, 0.6 g, 12.4998 mmol/2 eq) was added at 0 °C and stirred for 15 min. MeI (1.18 mL, 19.2306 mmol, 1.3 eq) was added dropwise to the RM at 0 °C. The RM was brought to RT and stirred for 5 h. After the reaction was complete, the RM was quenched in cold water and extracted with ethyl acetate. The combined organic fractions were dried over Na2SO4, concentrated, and then purified by flash column chromatography (15% ethyl acetate in hexanes) to give compound (71_Int-1). MS (ES): 274 m/z [M+1]+, 1H NMR (400 MHz, DMSO-d6) δ 1.267 (t, J = 7.2 Hz, 3H), 3.268 (s, 3H), 4.235-4.182 (m, 2H), 4.382 (s, 2H), 5.536 (s, 2H), 7.248 (t, J=8 Hz 4H), 7.877 (s, 1H), 8.468 (s, 1H). Step -2. Synthesis of N-(4- carbamimidoylbenzyl )-1-(4-( methoxymethyl ) benzyl )-1H- pyrazole -4- carboxamide (71) .

The final compound was prepared from ethyl 1-(4-(methoxymethyl)benzyl)-1H-pyrazole-4-carboxylate (71-Int-1) and 4-(aminomethyl)benzamididine dihydrochloride in a manner similar to that described in (Example-24) (71). MS (ES): 378.28 m/z [M+1]+, LCMS purity: 100%, HPLC purity: 98.18% 1H 400 MHz, DMSO-d6: δ 3.275 (s, 3H), 4.390 (s, 2H), 4.487 (d, J = 6.0 Hz, 2H), 5.353 (s, 2H), 7.248-7.318 (m, 4H), 7.50 (d, J =8.0 Hz, 2H), 7.758 (d, J =8.0 Hz, 2H), 7.924 (s, 1H), 8.281 (s, 1H), 8.791 (t, J=6.0 Hz, 1H), 9.074 (s, 2H), 9.264 (s, 2H). Example 34 : N-(4- Carbamimidoylbenzyl )-1-(4-( cyanomethyl ) benzyl )-5 -cyclopropyl -1H- pyrazole -4- carboxamide (86) Step -1. Synthesis of 2-(4-( cyanomethyl ) benzyl ) hydrazine -1- carboxylic acid tributyl ester (86_Int-2)

A stirred solution of 2-(4-(bromomethyl)phenyl)acetonitrile (86_Int-1) (3 g, 14.2857 mmol, 1.0 eq) in dimethylacetamide (30 mL, 10V) was prepared at RT. Then DIPEA (3.68 g, 28.5714 mmol, 2.0 eq) and tri-butyl hydrazinecarboxylate (3.77 g, 28.5714 mmol, 2.0 eq) were added at RT. The RM was then heated to 70 °C and stirred for 16 h. After the reaction was complete, the RM was quenched in DM water and extracted with ethyl acetate. The combined organic fractions were washed with saline solution, dried over Na2SO4, concentrated and purified by flash column chromatography (10-14% ethyl acetate in hexane) to give compound (86_Int-2) MS (ES): 261 m/z [M+1]+, LCMS purity: 70%. Step -2. Synthesis of 2-(4-( hydrazinomethyl ) phenyl ) acetonitrile (86_Int-3)

A stirring solution of tributyl 2-(4-(cyanomethyl)benzyl)hydrazine-1-carboxylate (86_Int-2) (1.0 g, 7.6628 mmol, 1.0eq) in methanol (10 mL, 5V) and water (22 mL, 11V) was prepared. Then concentrated HCl (6.6 mL, 3.3V) was added to the RM at RT. The RM was heated to 80°C and stirred for 16h. After completion of the reaction, the RM was evaporated and triturated in ethyl acetate to give compound (86_Int-3) MS (ES): 162.3 m/z [M+1]+, LCMS purity: 64%. Step -3. Synthesis of ethyl 1-(4-( cyanomethyl ) benzyl )-5 -cyclopropyl -1H- pyrazole -4- carboxylate (86_Int-4)

A solution of ethyl 3-cyclopropyl-3-oxopropanoate (1.0 g, 4.61531 mmol, 1.0 eq) and dimethylformamide dimethyl acetal (0.6 g, 4.2307 mmol, 1.1 eq) was prepared and stirred at 75 °C for 90 min. The RM was cooled to RT and then ethanol (12 mL, 20V), TEA (2.0 mL, 15.3844 mmol, 4.0 eq), and 2-(4-(hydrazinomethyl)phenyl)acetonitrile (86_Int-3) (0.600 g, 3.8461 mmol, 1.0 eq) were added. The RM was heated to 80 °C and stirred for 16 h. After the reaction was complete, the RM was quenched into water and extracted with ethyl acetate. The combined organic fractions were washed with saline solution, dried over Na2SO4, concentrated, and then purified by flash column chromatography (12-15% ethyl acetate in hexanes) to give compound (86_Int-4). MS (ES): 310 m/z [M+1]+, LCMS purity: 64%. 1H 400 MHz, DMSO-d6: δ 0.754 (d, J = 6.40 Hz, 2H), 0.963 (q, J = 6.4 Hz, 2H), 1.790-1.756 (m, 1H), 4.106 (s, 2H), 4.205-4.152 (m, 2H), 5.459 (s, 2H), 7.166 (d, J = 8 Hz, 2H), 7.240 (d, J = 8 Hz 2H), 7.814 (s, 1H). Step -4. Synthesis of N-(4 -carbamimidoylbenzyl )-1-(4-( cyanomethyl ) benzyl )-5 -cyclopropyl -1H- pyrazole -4 -carboxamide (86)

The final compound was prepared from ethyl 1-(4-(cyanomethyl)benzyl)-5-cyclopropyl-1H-pyrazole-4-carboxylate (86_Int-4) and 4-(aminomethyl)benzamididine dihydrochloride in a manner similar to that described in (Example-24) (86). MS (ES): 412.5 m/z [M+1]+, LCMS purity: 100%, HPLC purity: 100% 1H 400 MHz, DMSO-d6: δ 0.754 (d, J = 6.40 Hz, 2H), 0.963(q, J = 6.4 Hz, 2H), 1.790-1.756 (m, 1H), 4.025 (s, 2H), 4.487 (d, J=6.4 Hz, 2H), 5.440 (s, 2H),7.196 (d, J = 8 Hz, 2H), 7.33 (d, J=8 Hz, 2H), 7.522 (d, J= 8 Hz 2H), 8.538 (d, J=6.4 Hz, 2H), 8.964 (s, 2H), 9.266 (s, 2h). Example 35 : N-(4 -carbamimidoylbenzyl )-1-(4-(2- methoxyethyl ) benzyl )-5-( methoxymethyl )-1H- pyrazole -4 -carboxamide (87) Step -1. Synthesis of methyl 1-(4-(2- hydroxyethyl ) benzyl )-3-( methoxymethyl )-1H- pyrazole -4- carboxylate (87_Int-2)

Prepare a stirred solution of methyl 5-(methoxymethyl)-1H-pyrazole-4-carboxylate (3.2 g, 1.88235 mmol, 1.0 eq) in DMF (32 mL, 10 V) and add NaHMDS (1M in THF) (18.88 mL, 1.88235 mmol, 1.0 eq) at 0 °C and stir for 30 min. Add 2-(4-(bromomethyl)phenyl)ethan-1-ol (87_Int-1) (4.0 g, 1.88235 mmol, 1.0 eq) to the RM at 0 °C. Bring the RM to RT and stir for 16 h. After the reaction is complete, quench the RM into water and extract with ethyl acetate. The combined organic fractions were dried over Na2SO4, concentrated, and then purified by flash column chromatography (5% ethyl acetate in hexanes) (87_Int-2). Confirmed by 2D NMR (ROE analysis). MS (ES): 304.9 m/z [M+1]+, LCMS purity: 99.72%, 1H 400 MHz, DMSO-d6: δ 2.679 (t, J=4 Hz, 3H), 3.242(s, 3H), 3.555 (t, J=4 Hz, 2H), 3.724(s, 3H), 4.490(s, 2H), 5.276 (s, 2H), 7.203-7.184 (m, 4H), 8.416 (s, 1H). Step -2. Synthesis of methyl 1-(4-(2- methoxyethyl ) benzyl )-5-( methoxymethyl )-1H- pyrazole -4- carboxylate (87_Int-3)

A stirred solution of methyl 1-(4-(2-hydroxyethyl)benzyl)-5-(methoxymethyl)-1H-pyrazole-4-carboxylate (87_Int-2) (0.4 g, 1.3157 mmol, 1.0 eq) in DMF (4 mL, 10 V) was prepared. NaH (60% in mineral oil) (0.126 g, 2.6314 mmol, 2 eq) was added slowly and the RM was stirred at 0 °C for 15 min, followed by MeI (0.106 mL, 1.7106 mmol, 1.3 eq) added dropwise. The RM was brought to RT and stirred for 5 h. After completion of the reaction, the RM was quenched into cold water and extracted with ethyl acetate. The combined organic fractions were dried over Na2SO4, concentrated, and then purified by flash column chromatography (15% ethyl acetate in hexanes) to give compound (87_Int-3). MS (ES): 319 m/z [M+1]+, LCMS purity: 79%, 1H NMR (400 MHz, DMSO-d6) δ 400 MHz, DMSO-d6: δ 2.489 (t, J = 6.40 Hz, 2H), 3.211 (s, 3H), 3.225 (s, 3H), 3.518 (t, J=6.4 Hz, 2H), 4.794 (s, 2H), 5.358 (s, 2H), 7.111 (d, J=8 Hz, 2H), 7.200 (d, J=8 Hz, 2H), 7.905 (s, 2H). Step -3. N-(4 -carbamimidoylbenzyl )-1-(4-(2- methoxyethyl ) benzyl )-5-( methoxymethyl )-1H- pyrazole -4- carboxamide (87)

The final compound was prepared from methyl 1-(4-(2-methoxyethyl)benzyl)-5-(methoxymethyl)-1H-pyrazole-4-carboxylate (87_Int-3) and 4-(aminomethyl)benzamididine dihydrochloride in a similar manner to that described in (Example-24) (87). MS (ES): 435.53 m/z [M+1]+, LCMS purity: 100%, HPLC purity: 100% 1H 400 MHz, DMSO-d6: δ 2.489 (t, J = 6.40 Hz, 2H), 3.211 (s, 3H), 3.225 (s, 3H),3.503(t, J=6.4 Hz, 2H), 4.492 (d, J=6 Hz, 2H), 4.802(d, 2H), 5.310 (d, 2H), 7.111 (d, J=8 Hz, 2H), 7.183 (d, J=8 Hz, 2H), 7.508 (d, J=8 Hz, 2H), 7.752 (d, J=8 Hz, 2H), 2H). 8.012 (s, 1H), 8.817 (t, J=6 Hz, 1H), 8.911 (s, 2H), 9.252 (s, 2H). Example 36 : Preparation of N-(4 -carbamimidoylbenzyl )-1-(4-(2-( dimethylamino )-2- oxoethyl ) benzyl )-1H- pyrazole -4- carboxamide (33 ) Step -1. Synthesis of N-(4- carbamimidoylbenzyl )-1-(4-(2-( dimethylamino )-2- oxoethyl ) benzyl )-1H- pyrazole -4- carboxamide (33)

The final compound was prepared from ethyl 1-(4-(2-(dimethylamino)-2-oxoethyl)benzyl)-1H-pyrazole-4-carboxylate (33-Int-3) and 4-(aminomethyl)benzamidinium dihydrochloride in a similar manner as that described in (Example-24) (33). MS (ES): 419.4 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 95.56% 1H NMR (400 MHz, DMSO-d6) δ 2.807 (s, 3H), 2.978 (s, 3H), 3.656 (s, 2H), 4.778 (d, J=6.0 Hz, 2H), 7.183-7.197 (m, 5H), 7.483-7.504 (m, 2H), 7.734-7.755 (m, 1H), 7.837 (s, 1H), 7.907 (s, 1H), 8.269 (s, 1H), 8.767 (t, J=6.0 Hz, 1H), 8.946 (s, 2H), 9.245 (s, 2H). Example 37 : Preparation of N-(4- carbamimidoylbenzyl )-1-(2-( cyanomethyl ) benzyl )-1H -pyrazole -4- carboxamide (70) Step -1. Synthesis of ethyl 1-(2-( cyanomethyl ) benzyl )-1H- pyrazole -4- carboxylate (70_Int-2)

A stirred solution of ethyl 1H-pyrazole-4-carboxylate (0.1 g, 0.71 mmol, 1.0 eq) in DMF (2 mL, 20 V) was prepared and 2-(2-(bromomethyl)phenyl)acetonitrile (70_Int-1) (0.149 g, 0.71 mmol, 1.0 eq) was added at RT. The RM was then heated to 80 °C and stirred for 16 h. After the reaction was complete, the RM was quenched in water and extracted with ethyl acetate. The combined organic fractions were dried over Na2SO4, concentrated, and purified by flash column chromatography (25% ethyl acetate in hexanes) to give the compound. MS (ES): 270.m/z [M+1]+, LCMS purity: 98.57%, 1H 400 MHz, DMSO-d6: δ 1.244 (t, J = 6.4 Hz, 3H), 4.225-4.172 (m, 4H), 5.448 (s, 2H), 7.119 (d, J = 7.20 Hz, 1H), 7.454-7.332 (m, 3H), 7.892 (s, 1H), 8.453 (s, 1H). Step -2. Synthesis of N-(4- carbamimidoylbenzyl )-1-(2-( cyanomethyl ) benzyl )-1H- pyrazole -4- carboxamide (70)

The final compound was prepared from ethyl 1-(2-(cyanomethyl)benzyl)-1H-pyrazole-4-carboxylate (70_Int-2) and 4-(aminomethyl)benzamididine dihydrochloride in a similar manner as described in (Example-24) (70). MS (ES): 373.45 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 98.64. 1H 400 MHz, DMSO-d6: δ 4.185 (s, 2H), 4.483 (d, J =6.0 Hz, 2H), 5.444 (s, 2H), 7.119 (s, 1H), 7.345-7.396 (m, 2H), 7.448-7.504 (m, 3H), 7.746 (d, J=8.0 Hz, 2H), 7.943 (s, 1H), 8.259 (s, 1H), 8.784 (t, J=6.0 Hz, 1H), 8.952 (s, 2H), 9.247 (s, 2H). Example 38 : Preparation of N-(4- carbamimidoyl -3- fluorobenzyl )-1-(4-(2-( methylamino )-2- oxoethyl ) benzyl )-1H- pyrazole -4- carboxamide (74) Step -1. Synthesis of N-(4- carbamimidoyl -3- fluorobenzyl )-1-(4-(2-( methylamino )-2- oxoethyl ) benzyl )-1H- pyrazole -4- carboxamide (74)

The final compound was prepared from ethyl 1-(4-(2-(methylamino)-2-oxoethyl)benzyl)-1H-pyrazole-4-carboxylate (19-Int-3) and 4-(aminomethyl)-2-fluorobenzamidine dihydrochloride (43-Int-6) in a manner similar to that described in (Example-24) (74). MS (ES): 423.4 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 98.51% 1H NMR (400 MHz, DMSO-d6) δ 2.678 (s, 3H), 3.367 (s, 2H), 4.467 (d, J=6 Hz, 2H), 5.317 (s, 2H), 7.196-7.243 (m, 4H), 7.328 (t, J=8.4 Hz, 2H), 7.262 (t, J=8.4 Hz, 1H), 7.906 (s, 1H), 7.961 (s, 1H), 8.268 (s, 1H), 8.807 (t, J=6.0 Hz, 1H), 9.186 (s, 2H), 9.387 (s, 2H). Example 39 : Preparation of N-(4 -carbamimidoylbenzyl )-1-(4-(2- cyanoprop -2- yl ) benzyl )-5-( methoxymethyl )-1H- pyrazole -4 - carboxamide (82) Step -1. Synthesis of N-(4- carbamimidoylbenzyl )-1-(4-(2- cyanoprop -2- yl ) benzyl )-5-( methoxymethyl )-1H- pyrazole -4- carboxamide (82)

The final compound was prepared from methyl 1-(4-(2-cyanopropan-2-yl)benzyl)-5-(methoxymethyl)-1H-pyrazole-4-carboxylate (82-Int-3) and 4-(aminomethyl)benzamidinium dihydrochloride in a similar manner as described in (Example-24) (70). MS (ES): 445.4 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 100% 1H NMR (400 MHz, DMSO-d6) δ 1.675 (s, 6H), 3.246 (s, 3H), 4.514 (d, J=6 Hz, 2H), 4.480 (s, 2H), 5.380 (s, 2H), 7.274 (d, J=8.4 Hz, 2H), 7.486-7.537 (m, 4H), 7.777 (d, J=8.4 Hz, 2H), 8.046 (s, 1H), 8.843 (t, J=6 Hz, 1H), 9.022 (s, 2H), 9.274 (s, 2H). Example 40 : Preparation of N-(4- carbamimidoylbenzyl )-1-(4-(2- cyanopropan -2- yl ) benzyl )-5-( methoxymethyl )-1H- pyrazole -4- carboxamide (85) Step -1. Synthesis of N-(4- carbamimidoylbenzyl )-1-(4-( cyanomethyl ) benzyl )-5-( methoxymethyl )-1H- pyrazole -4- carboxamide (85)

The final compound was prepared from methyl 1-(4-(cyanomethyl)benzyl)-5-(methoxymethyl)-1H-pyrazole-4-carboxylate (85_Int-3) and 4-(aminomethyl)benzamididine dihydrochloride in a similar manner as described in (Example-24) (70). MS (ES): 417.4 m/z [M+H]+, LCMS purity: 100%, HPLC purity: 100% 1H NMR (400 MHz, DMSO-d6) δ 3.233 (s, 3H), 4.029 (s, 2H), 4.514 (d, J=6 Hz, 2H), 4.829 (s, 2H), 5.375 (s, 2H), 7.232 (d, J=8 Hz, 2H), 7.327 (d, J=8 Hz, 2H), 7.529 (d, J=8.0 Hz, 2H), 7.774 (d, J= 8.0 Hz, 2H), 8.046 (s, 1H), 8.839 (t, J=6 Hz, 1H), 8.963 (s, 2H), 9.271 (s, 2H). Example 41 : Preparation of N-(4 - carbamimidoyl -3- fluorobenzyl )-1-(4-(2-( dimethylamino )-2 -oxoethyl ) benzyl )-1H- pyrazole -4- carboxamide (78) Step -1. Synthesis of 1-(4-(2-( dimethylamino )-2 -oxoethyl ) benzyl )-1H- pyrazole -4- carboxylic acid (78_Int-1)

A stirred solution of ethyl 1-(4-(2-(dimethylamino)-2-oxoethyl)benzyl)-1H-pyrazole-4-carboxylate (33_Int-3) (0.5 g, 0.158 mmol, 1.0 eq) in water:methanol:THF (1:1:1) (5 mL, 10 V) was prepared and LiOH (0.3 g, 0.793 mmol, 5 eq) was added at RT and stirred for 16 h. After completion of the reaction, the RM was quenched in 2N HCl (approximately pH 4) and extracted with 10% methanol in DCM. The combined _organic fractions were dried over _Na2SO4 and concentrated to give compound (78_Int-1) MS (ES): 288.4.m/z [M+H]+, LCMS purity: 97.47%, 1H NMR (400 MHz, DMSO-d6) δ 2.804 (s, 3H), 2.975 (s, 3H), 5.315 (s, 2H), 7.191-7.219 (m, 4H), 7.795 (s, 1H), 8.357 (s, 1H), 12.301 (s, 1H). Step -2. Synthesis of N-(4- carbamimidoyl -3- fluorobenzyl )-1-(4-(2-( dimethylamino )-2- oxoethyl ) benzyl )-1H- pyrazole -4- carboxamide (78)

A stirred solution of 1-(4-(2-(dimethylamino)-2-oxoethyl)benzyl)-1H-pyrazole-4-carboxylic acid (78_Int-1) (0.2 g, 0.069 mmol, 1.0 eq) in pyridine (2 mL, 10 V) was prepared. -(aminomethyl)-2-fluorobenzamidine dihydrochloride (43_Int-5) (0.2 g, 0.083 mmol, 1.2 eq) and EDC HCl (0.67 g, 0.348 mmol, 5.0 eq) were added to the RM at RT and stirred for 16 h. After the reaction was complete, the RM was concentrated and purified by Prep HPLC ((A) 0.1% TFA in water (B) 100% MeCN). The pure fractions were lyophilized to give compound (78) MS (ES): 437.36.m/z [M+H]+, LCMS purity: 98.37%, HPLC purity: 99.70% 1H NMR (400 MHz, DMSO-d6) δ 2.804 (s, 3H), 2.981 (s, 3H), 3.653 (s, 2H), 4.459 (d, J=6.4 Hz, 2H), 5.314 (s, 2H), 7.194 (s, 4H), 7.318 (t, J=11.6 Hz, 2H), 7.615 (t, J=11.6 Hz, 1H), 7.900 (s, 1H), 8.266 (s, 1H), 8.790 (t, J=6 Hz, 1H), 9.170 (s, 2H), 9.370 (s, 2H). Biological examples

Plasma Microangiotensin Protease Inhibition Assay . Two methods are provided to determine the IC ₅₀ of test compounds against plasma microangiotensin.

In the first method, a reaction buffer was used, which contained 25 mM Tris-HCl (pH 8.0), 100 mM NaCl (pH 8.5), 0.01% Brij35, and 1% DMSO (final). The enzyme used was plasma microangiotensin (R&D Systems Cat# 2497-SE; recombinant human plasma microangiotensin expressed in a mouse myeloma cell line, NS0-derived Gly20-Ala638 with a C-terminal 60-His tag. MW = 70 kDa).

The enzyme was activated by diluting to 200 µg/mL in activation buffer (100 mM Tris, 10 mM CaCl2, 150 mM NaCl, pH 7.5 (TCN)) and then combined with an equal volume of 20 µg/mL thermolysin to form the reaction buffer. Each test compound was then dissolved in DMSO and delivered to the reaction buffer. After a 20-minute preincubation period, the reaction was initiated by delivering a substrate solution containing 10 µM Z-FR-AMC (Enzo Cat# P-139; AMC: 7-amino-4-methylcoumarin) to the reaction wells.

Measurements were performed using EnVision (PE) with excitation and emission wavelengths of 355 nm and 460 nm, respectively. Reactions were stopped using EDTA. Enzyme activity was monitored every 5 minutes for 120 minutes at room temperature as a time course measurement of the increase in signal from the fluorescently labeled peptide substrate.

Data were analyzed by taking the slope*(signal/time) of the linear portion of the measurements. The slope was calculated using Excel and the curve fit was performed using Prism software.

Secondly, plasma microangiotensin activity was also measured in pooled human plasma. First, a 10% Actin FS solution was prepared in assay buffer. Each test compound was dissolved in DMSO and delivered to the reaction mixture together with the Z-FR-AMC substrate and pooled human plasma. The multiwell reaction plate was incubated at room temperature for five minutes. To start the reaction, a 10% Actin FS solution was added to each well and kinetic measurements were performed at Ex/Em at 355/460 nm. The fluorescence signal was recorded every 30 seconds for a total of 10 minutes.

Permeability assay. Permeability studies were performed using Caco-2 cells (ECACC Cat. no. 09042001) seeded at a density of 80,000 cells per well in cell culture plates and maintained in culture medium (1x DMEM with 10% FBS, 0.1 mg/mL penicillin/streptomycin) for 18 to 21 days. The culture medium was changed every other day. Prior to the experiment, the integrity of the cell monolayer was assessed by measuring the TEER value using a voltohmmeter and a STX100C96 electrode. Only monolayers with a TEER value higher than 800 ohm.cm2 in buffer after the first wash were used.

Assay buffer (HBSS with Ca+2 and Mg+2, buffered with 10 mM HEPES and 25 mM D-Glucose, pH -7.4) was used on the apical side and on the basolateral side.

An intermediate stock solution of the test compound was prepared in DMSO at a concentration of 1 mM in DMSO. This stock solution was spiked into the assay buffer to achieve a target test compound concentration of 10 µM. The organic content of the final drug formulation was 1.0% v/v. Two-way permeability experiments were performed in single runs and sample analysis was performed in duplicate.

The cultured cell monolayer was washed with assay buffer (0.4 mL and 0.8 mL were added to the apical and basolateral sides of the culture plate, respectively) and the buffer from both compartments was discarded.

For apical to basolateral (AP>BL) experiments, aliquots of 0.4 mL of donor solution (assay buffer, pH 7.4, containing test compounds) and 0.8 mL of receptor solution (assay buffer only, pH 7.4) were added to the apical and basolateral compartments, respectively. For basolateral to apical (BL>AP) experiments, aliquots of 0.8 mL of donor solution (assay buffer, pH - 7.4, containing test compounds) and 0.4 mL of receptor solution (assay buffer, pH - 7.4) were added to the basolateral and apical compartments, respectively. The plates were then kept in an incubator at 37°C for 120 min.

Control experiments for these two directions (AP＞BL and BL＞AP) with Propranolol (high permeability), Atenolol (low permeability), Digoxin (high efflux-Pgp substrate), and Digoxin+Verapamil (Pgp inhibitor) were performed in separate wells on the same day of the experiment.

After completion of the transport experiment, the integrity of the cell monolayer was assessed by measuring Lucifer Yellow (LY) exclusion. To this end, 400 µL of 10 µM LY was added to each well of the filter disk and incubated at 37°C for 1 hour. Subsequently, samples were collected from the basolateral compartment and LY fluorescence was measured using an excitation wavelength of 485 nm and an emission wavelength of 530 nm. The percentage of LY exclusion across the cell monolayer was assessed by measuring the fluorescence in the receptor disk (basolateral compartment) compared to a theoretical equilibrium standard.

Study samples (collected from apical and basolateral compartments after 120 min incubation) were analyzed by LCMS/MS, and Papp was calculated for the compound(s) in both apical-to-basolateral and basolateral-to-apical directions. Papp = ([DBL] x VBL)/(A × t × [DAP]), where: [DBL] = final drug concentration on the basolateral side, VBL = volume of the basolateral compartment, A = surface area of cell culture, t = total incubation time, and [DAP] = initial drug concentration on the apical side.

The results from the above assays for representative compounds of the present disclosure are presented in Table 1 below. The scores of the selected compounds in each assay are presented as follows: Purified human plasma assay (nM) Storage human plasma assay (nM) Caco-2 permeation assay (10 ^-6 cm/s) A ＜ 1 A ＜ 100 A ＜ 2 1 ≤ B ＜ 10 100 ≤ B ＜ 1000 2 ≤ B ＜ 10 10 ≤ C ＜ 100 1000 ＜ C 10 ＜ C 100 ＜ D Table 1. Inhibition of plasma microangiotensin and cell permeation by representative compounds Examples Structure Purified PKal IC50 (nM) Storage human plasma PKal IC50 (nM) Caco-2 Perm (10 ^-6 cm/s) 1 A A A 2 B A 3 A A A 4 A A 5 B 6 7 A A A 8 B A 9 B B 10 B 11 B 12 C B 13 A A 14 A B 15 A A B 16 D 17 18 B A 19 A A A 20 twenty one twenty two twenty three twenty four 25 A A 26 A A A 27 28 A A A 29 30 31 32 33 A A A 34 35 36 37 38 39 40 41 42 43 B A A 44 D C 45 A A B 46 47 48 C B A 49 A B A 50 A A A 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 B A 70 C B 71 A A 72 C B 73 B A 74 B B 75 D C 78 81 A A 82 C A 83 C B 84 D D 85 C B 86 C B 87 C A A

It should be understood that the examples and embodiments described herein are for illustrative purposes only and that persons of ordinary skill in the art will conceive of various modifications or variations thereof and incorporate such modifications or variations within the spirit and scope of this application and the scope of the appended patent applications. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.

without

without

Claims (31)

A compound of formula (IA) or a pharmaceutically acceptable salt thereof: (IA) wherein ^D1 is N or ^CR1 , ^D2 is N or ^CR2 , ^D3 is N or ^CR3 , and ^D4 is N or ^CR4 , wherein no more than three of ^D1 , ^D2 , ^D3 , and ^D4 are simultaneously N; ^R1 , ^R2 , ^R3 , and ^R4 are independently selected from the group consisting of H, _C2 - _C6 -alkenyl, _C1 - _C6 -halogenalkyl, halogen, ^NRaRb , ^ORa , ^-NRaC (O) ^Rb , -C(O) ^Ra , ^-C (O)halogen, -OC(O) ^Ra , -OC(O) ^ORa , -C(O) ^ORa , _C6 -C10 _- aryl, CN, -S(O) _0-2Ra ^{, -S(O)2ORa} _, and NO ₂ ; each of ^Ra and ^Rb is independently selected from the group consisting of H, C ₁ -C ₆ -alkyl, and C ₁ -C ₆ -haloalkyl; ^Rc1 , ^Rc2 , and ^Rc3 are independently selected from the group consisting of H, C ₁ -C ₆ -alkyl, and C ₁ -C ₆ -haloalkyl; _Q1 , _Q2 , and _Q3 are independently selected from the group consisting of ^CR5 , N, O, and S; ^R5 is selected from the group consisting of H, C ₁ -C ₆ -alkyl, ^ORa , -(C ₁ -C ₆ -alkyl) ^ORa , and C ₃ -C ₁₀ -cycloalkyl; P is C or N; ^L1 is _-SO2- or -C ₁ -C ₈ -alkylene-; is a divalent monocyclic or bicyclic moiety selected from the group consisting of: C ₃ -C ₁₀ -cycloalkyl, C ₆ -C ₁₀ -aryl, 3 to 10-membered heterocycloalkyl (wherein 1 to 4 ring members are independently selected from N, O, and S), 5 to 10-membered heteroaryl (wherein 1 to 4 heteroaryl members are independently selected from N, O, and S), and fused combinations thereof; L ² is selected from the group consisting of: -C ₁ -C ₈ -alkylene, -C ₂ -C ₈ -alkenylene, -C ₂ -C ₈ -alkynylene, and a divalent moiety selected from the group consisting of: C ₃ -C ₁₀ -cycloalkyl, C ₃ -C ₁₀ -cycloalkenyl, C ₆ -C ₁₀ -aryl, 5- to 10-membered heteroaryl (wherein 1 to 4 heteroaryl members are independently selected from N, O, and S), (C ₁ -C ₈ -alkyl)C ₃ -C ₁₀ -cycloalkyl, (C ₁ -C ₈ -alkyl)C ₃ -C ₁₀ -cycloalkenyl, (C ₁ -C ₈ -alkyl)C ₆ -C ₁₀ -aryl, and (C ₁ -C ₈ -alkyl)5- to 10-membered heteroaryl (wherein 1 to 4 heteroaryl members are independently selected from N, O, and S), wherein cycloalkyl, cycloalkenyl, aryl, and heteroaryl are monocyclic or bicyclic; wherein L ² is optionally substituted with 1 to 3 substituents selected from the group consisting of C ₁ -C ₆ -alkyl, C ₁ -C ₆ -haloalkyl, and CN; Z is selected from the group consisting of ^-ORc , -OC(O) ^Rc , -OC(O) ^{NRcRd ,} -S(O) _0-2Rc , ^-CN , -C(O) ^Rc , ^-C (O)ORc, -C(O) ^{NRcRd ,} -C(S) ^Rc _, ^-NRcRd , =NRc, ^{-NRcC (} O) ^{NRcRd , -NRcCO2Rd , -NRc} -NO, _-NO2 , ^-NRc - ^ORd , -N=C=O, -N= ^C =S, and ^-NRc - ^NRcRd ; and each example ^{of Rc and Rd is independently} selected from H, _C1 - _C6 -alkyl, _C1 - _C6 -haloalkyl, and _C6 - _C10 -aryl. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein no more than one of D ¹ , D ² , D ³ , and D ⁴ is N. The compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein the ring containing D ¹ , D ² , D ³ , and D ⁴ is selected from the group consisting of: , ,and . The compound of any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, wherein the ring system containing D ¹ , D ² , D ³ , and D ⁴ is . The compound of any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² , R ³ , and R ⁴ are independently selected from the group consisting of H, OR ^a , halogen, and CN. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein R ¹ , R ² , R ³ , and R ⁴ are independently selected from the group consisting of H and halogen. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein at least one of R ¹ , R ² , R ³ , and R ⁴ is a halogen group. The compound of any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, wherein each of R ¹ , R ² , R ³ , and R ⁴ is H. The compound of any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein each of R ^c1 , R ^c2 , and R ^c3 is H. The compound or pharmaceutically acceptable salt thereof of any one of claims 1 to 9, wherein ^Q1 is C; each of ^Q2 and ^Q3 is independently selected from the group consisting of ^CR5 , N, O, and S; and P is selected from the group consisting of C and N. The compound of any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof, wherein P is N. The compound of any one of claims 1 to 11 or a pharmaceutically acceptable salt thereof, wherein Q ² is CR ⁵ and Q ³ is N. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, wherein Q ² is N and Q ³ is CR ⁵ . The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 13, wherein L ¹ is -C ₁ -C ₈ -alkylene-. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 14, wherein L ¹ is -C ₁ -C ₃ -alkylene-. A compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof, wherein It is a divalent monocyclic C ₆ -C ₁₀ -aryl group. A compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof, wherein Department . The compound of any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof, wherein L ² is selected from -C ₁ -C ₈ -alkylene, C ₆ -C ₁₀ -aryl, and (C ₁ -C ₈ -alkyl)C ₆ -C ₁₀ -aryl. The compound of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, wherein L ² is -C ₁ -C ₈ -alkylene. The compound of any one of claims 1 to 19 or a pharmaceutically acceptable salt thereof, wherein L ² is C ₁ -C ₃ -alkylene. The compound of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, wherein L ² is C ₆ -C ₁₀ -aryl. The compound or pharmaceutically acceptable salt thereof of any one of claims 1 to 21, wherein L ² is phenyl. The compound or pharmaceutically acceptable salt thereof of any one of claims 1 to 22, wherein Z is selected from the group consisting of -OR ^c , CN, -C(O)OR ^c , and -C(O)NR ^c R ^d . The compound of any one of claims 1 to 23 or a pharmaceutically acceptable salt thereof, wherein Z is CN. The compound of any one of claims 1 to 23 or a pharmaceutically acceptable salt thereof, wherein each of R ^c and R ^d is independently H or C ₁ -C ₆ -alkyl. The compound or pharmaceutically acceptable salt thereof of any one of claims 1 to 24, wherein Z is selected from the group consisting of OH, OCH ₃ , -COOH, -C(O)NH ₂ , and -C(O)NHCH ₃ . The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has formula (I): (I) wherein Q ¹ , Q ² , and Q ³ are selected from the group consisting of C, N, O, and S; P is selected from the group consisting of C and N; L ¹ is a linker selected from the group consisting of C ₁ -C ₈ alkyl linkers and SO ₂ ; i) a cyclic hydrocarbon, bicyclic hydrocarbon, or heterocyclic hydrocarbon containing up to 10 atoms consisting of C or N; or ii) selected from the group consisting of cyclopentyl, cyclohexyl, phenyl, naphthyl, pyridine, pyrimidine, indene, 2,3-dihydro- 1H -indene, or any saturated and unsaturated cyclic hydrocarbon or heterocyclic hydrocarbon thereof; ^L2 is i) a hydrocarbon not containing a double bond to O or S, ii) a hydrocarbon and not containing heteroatoms such as O, N, or S; or iii) a bonding group selected from the group consisting of a _C1 -C2-containing hydrocarbon, an unsaturated hydrocarbon, a branched hydrocarbon, a cyclic hydrocarbon, and combinations thereof; ₈ alkyl bond linker; a cyclic hydrocarbon selected from the group consisting of cyclopentyl, cyclohexyl, phenyl, naphthyl, indene, and 2,3-dihydro- 1H -indene, or any saturated or unsaturated cyclic hydrocarbon thereof; Z is i) selected from the group consisting of -OH, -OR', -OC(O)H, -OC(O)R', -OC(O) _NH2 , -OC(O)NHR', -OC(O)NH(R') ₂ , -SH, -SR', -S(O)R', -S(O) _2R ', -CN, -C(O)H, -C(O)R', -C(O)OH, -C(O)OR', -C(O) _NH2 , -C(O)NHR', -C(O)NH(R') ₂ , -C(S)R', _-NH2 , _-NH2R ', -NR'2, =NH _, =NR', -NHC(O) _NH2 , -NR'C(O)NH2, -NR'C(O)NHR', _-NR'C (O)N(R') ₂ , -NHCO2H _{, -NHCO2R} ', -NR'CO2R', -NH-NO, -NR' _- NO, _-NO2 , -NH-OH, -OH, -NR'-OR', -N=C=O, -N=C=S, -NH-NH2, and -NH-NHR', wherein each R' is independently alkyl or alkyl halide, or ii) is not halogen or hydrogen. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the following table: 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 twenty one twenty two twenty three twenty four 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 78 81 82 83 84 85 86 87 A pharmaceutical composition comprising the compound of any one of claims 1 to 28 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. A method for treating a subject suffering from a disease or condition, comprising administering to the subject a compound of any one of claims 1 to 28 or a pharmaceutically acceptable salt thereof, wherein the disease or condition is selected from the group consisting of ischemic stroke, hemorrhagic stroke, hypertension, retinopathy, diabetic retinopathy, nephropathy, cerebral edema suppression, pulmonary hypertension, inflammation, acute myocardial infarction, deep venous embolism, complications from fibrinolytic therapy, stroke, angina, vascular edema, Hyperemia, arthritis, complications of cardiopulmonary bypass, microvascular leak syndrome, inflammatory bowel disease, vascular complications from diabetes, diabetic macular edema, macular degeneration, neuropathy, age-related macular degeneration, retinal vein occlusion, cerebral edema, ischemia-reperfusion injury, angiogenesis, asthma, acute allergy, Alzheimer's disease, Parkinson's disease, multiple sclerosis, glioblastoma multiforme, complications of fibrinolytic therapy, increased albumin secretion, macroalbuminuria, pain , amyotrophic lateral sclerosis, Kugelman's disease, epilepsy, brain trauma, high altitude cerebral edema, cancer, generalized intravascular coagulation, pancreatitis, inflammation, shock, hereditary vascular edema (HAE), uveitis, polyangiitis, acute respiratory distress syndrome (ARDS), thrombosis, vasculitis, Crohn's disease, ulcerative colitis, colitis, arteritis, glomerulonephritis, eczema, endometriosis, preeclampsia, malaria, arthritis, intermittent and relapsing fever, Choker's disease, Raynaud's disease, systemic sclerosis, granulomatosis with polyangiitis, small vessel vasculitis, medium vessel vasculitis, large vessel vasculitis, pan vasculitis, systemic autoinflammatory disease, renal failure, cerebral malaria, Clarkson's disease (systemic vascular leak syndrome), Hantavirus infection, Hantavirus renal syndrome, Hantavirus pulmonary syndrome, virus-related inflammatory disorders, retinal vasculitis, uveitis, Ellis disease, Behcet's disease, sarcoidosis, pertussis, coronavirus infection, and noninfectious posterior uveitis. The method of claim 30, wherein the disease or condition is selected from the group consisting of diabetic macular edema, diabetic retinopathy, and uveitis.