WO2025111705A1 - Quinazolinone derivatives as poly (adp ribose) polymerase 1 (parp-1) inhibitors - Google Patents

Abstract

Disclosed is a compound of formula (I), its pharmaceutically acceptable salt or pro-drug thereof, where R¹, R², X, L¹, L², L³, L⁴, L⁵, and R are as disclosed herein. The compounds disclosed herein can have poly[ADP-Ribose) polymerase 1 (PARP- 1) inhibitory activity. As such, the compounds and compositions containing the compounds, as disclosed herein, can have use for treatment of diseases associated with PARP-1.

Description

QUINAZOLINONE DERIVATIVES AS POLY (ADP RIBOSE) POLYMERASE 1 (PARP-1) INHIBITORS

CROSS REFERENCE TO RELATED APPLICATION

[0001] This application claims the benefit of and priority to US Provisional Patent Application No.. 63/604245, having the title “POLY [ADP RIBOSE] POLYMERASE 1 (PARP-1) INHIBITORS”, filed on November 30^th, 2023. The content of the above-noted patent application is hereby expressly incorporated by reference into the detailed description hereof.

FIELD

[0002] The specification relates to Poly[ADP-Ribose) Polymerase 1 (PARP-1) inhibitors, their process for preparation and use thereof.

BACKGROUND

[0003] Cancer is a group of diseases involving abnormal cell growth with the potential to migrate to and affect other different parts of the body. Due to the broad range of diseases that are classified as cancer, a wide variety of different approaches are required for its treatment. Typical cancer treatments can include radiation therapy and chemotherapeutic drugs such as alkylating agents, anti-metabolites and topoisomerase inhibitors, which are designed to damage DNA, halt cellular division and influence apoptosis of cancer cells.

[0004] It is postulated that the efficacy of chemotherapy and radiation therapy is dampened by the activity of DNA repair enzymes (Ganesan, S. MYC, PARP1 , and Chemoresistance: BIN There, Done That? Sc/. Signal., 2011 , 4, 15, incorporated herein by reference). Two specific enzyme that are involved in the detection and repair of DNA damage is poly(ADP-ribose) polymerase-1 (PARP-1), and PARP-2 and these are members of the PARP family of enzymes (Herceg Z, Wang ZQ. Functions of poly(ADP-ribose) polymerase (PARP) in DNA repair, genomic integrity and cell death. Mutat Res. 2001 Jun 2;477(1-2):97-110, incorporated herein by reference). Inhibitors that target PARP enzymes and the mechanism of DNA repair are a new class of drugs that have recently received a large degree of attention and are of clinical importance with regard to various forms of cancer (Jagtap, P.; Szabo, C. Poly(ADP-ribose) polymerase and the therapeutic effects of its inhibitors, Nat. Rev. Drug Discov. 2005, 4, 421 ; Rouleau, M.; Patel, A.; Hendzel, M. J.; Kaufmann, S. H.; Poirier, G. G. PARP inhibition: PARP1 and beyond. Nat. Rev. Cancer, 2010, 10, 293; Cepeda, V.; Fuertes, M. A.; Castilla, J.; Alonso, C.; Quevedo, C;. Soto, M.; Perezb, M. Poly(ADP-Ribose) Polymerase-1 (PARP-1) inhibitors in cancer chemotherapy, Recent Patents on AntiCancer Drug Discovery, 2006, 1 , 39; Ferraris, D. V. Evolution of Poly(ADP-ribose) Polymerase-1 (PARP-1) Inhibitors. From Concept to Clinic. J. Med. Chem. 2010, 53, 4561 , all incorporated herein by reference).

[0005] Development of PARP-1 inhibitors has gone through several fluctuations. In the past, failure of several advanced clinical PARP-1 inhibitors dampened the enthusiasm for PARP inhibitors; however, it is notable that PARP-1 inhibitors have shown great potential to target cancers such as high-grade ovarian cancers and triple-negative breast cancers that are resistant to current treatment. Today, there are now a variety of PARP-1 inhibitors that have been approved by the FDA for the treatment of certain kinds of cancer including Olaparib, Talazoparib, and others (Rudolpha, J., Junga, K., Lugera, K. Inhibitors of PARP: Number crunching and structure gazing. Proc. Natl. Acad. Sci. U.S. A. 2022, 119 (11): 1):e2121979119, incorporated herein by reference).

[0006] There is still a need in the art for PARP-1 inhibitors. In addition, there is a need in the art for a process for preparation of PARP-1 inhibitors. Further, there is a need in the art for a composition containing PARP-1 inhibitors. Moreover, there is a need in the art for a method of treatment of a disease using PARP-1 inhibitors.

Furthermore, there is a need in the art for use of PARP-1 inhibitors for treatment of a disease.

[0007] The background herein is included solely to explain the context of the disclosure. This is not to be taken as an admission that any of the material referred to was published, known, or part of the common general knowledge as of the priority date.

SUMMARY

[0008] In one aspect, the specification relates to a compound of formula (I):

[0009] or a pharmaceutically acceptable salt or pro-drug thereof,

[0010] wherein

[0011] R¹ and R² independently are H, halogen, a Ci-6-substituent optionally having one or more heteroatoms, OR¹¹, or NR¹²R¹³;

[0012] R¹¹, R¹² and R¹³ are independently selected from H, benzyl, a

Ci-6-substituent optionally having with one or more heteroatoms, C(=O)R¹⁴, C(=O)OR¹⁵, or C(=O)NR¹⁶R¹⁷;

[0013] R¹⁴, R¹⁵, R¹⁶, and R¹⁷ are independently selected from H, benzyl, an optionally substituted aromatic ring, an optionally substituted heteroaromatic ring, or a Ci-6-substituent optionally having one or more heteroatoms;

[0014] X is O, S, or NH;

[0015] L¹ is absent or is NH, and when L¹ is absent, the quinazolinone moiety is bonded to L²;

[0016] L² is a Ci-3-chain optionally substituted with one or more methyl or methoxy, and L² being further optionally substituted with one or more heteroatoms;

[0017] L³ is absent or is NH, O, S;

[0018] L⁴ is absent or is - CH2 -;

[0019] L⁵ is absent or is C(=O), C(=S), S(=O) or S(=O)₂;

[0020] wherein when L³ is absent, L² is bonded to L⁴; when L³ and L⁴ are absent, L² is bonded to L⁵; when L³, L⁴ and L⁵ are absent, L² is bonded to R; when L⁴ is absent, L³ is bonded to L⁵; when L⁴ and L⁵ are absent, L³ is bonded to R; and when L⁵ is absent, L⁴ is bonded to R;

[0021] or [0022] L¹, L², L³, L⁴ and L⁵ together form a three to six membered ring optionally having one or more heteroatoms; and

[0023] R is H or a Ci-20-substituent optionally having one or more heteroatoms.

[0024] In a second aspect, the specification relates to a method for treating diseases mediated by PARP-1 protein comprising administering to a mammal in need thereof an effective amount of the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, or combination thereof, as disclosed herein.

[0025] In a third aspect, the specification relates to an in vitro method for inhibiting PARP-1 protein activity, comprising contacting the said protein with an effective amount of the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, or combination thereof, as disclosed herein.

[0026] In a fourth aspect, the specification relates to a pharmaceutical composition comprising a therapeutically effective amount of the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, or combination thereof, as disclosed herein, and at least one pharmaceutically acceptable excipient, carrier or diluent.

[0027] In a fifth aspect, the specification relates to a product comprising the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, or combination thereof, as disclosed herein, and one or more chemotherapeutic agents, as a combined preparation for simultaneous, separate or sequential use in anticancer therapy.

[0028] In a fifth aspect, the specification relates to the use of the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, or combination thereof, as disclosed herein, in the manufacture of a medicament for treatment of cancer.

[0029] In a sixth aspect, the specification relates to the use of the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, or combination thereof, as disclosed herein, for treating cancer. DESCRIPTION OF EXAMPLE EMBODIMENTS

[0030] Unless otherwise explained, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice for testing of the present invention, the typical materials and methods are described herein. In describing and claiming the present invention, the common terminology generally used is described herein below. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only, and is not intended to be limiting.

[0031] Many patent applications, patents, and publications are referred to herein to assist in understanding the aspects described. Each of these references are incorporated herein by reference in their entirety.

[0032] When introducing elements disclosed herein, the articles “a”, “an”, “the”, and “said” are intended to mean that there may be one or more of the elements.

[0033] The term "comprising" and its derivatives, as used herein, are intended to be open ended terms that specify the presence of the stated features, elements, components, groups, integers, and/or steps, but do not exclude the presence of other unstated features, elements, components, groups, integers and/or steps. The foregoing also applies to words having similar meanings such as the terms, "including", "having" and their derivatives. It will be understood that any embodiments described as “comprising” certain components may also “consist of” or “consist essentially of,” these components, wherein “consisting of” has a closed- ended or restrictive meaning and “consisting essentially of” means including the components specified but excluding other components except for materials present as impurities, unavoidable materials present as a result of processes used to provide the components, and components added for a purpose other than achieving the technical effects described herein. For example, a composition defined using the phrase “consisting essentially of” encompasses any known acceptable additive, excipient, diluent, carrier, and the like, suitable for the composition described herein. Typically, a composition consisting essentially of a set of components will comprise less than 5% by weight, typically less than 3% by weight, more typically less than 1 % by weight of non-specified components.

[0034] It will be understood that any component defined herein as being included may be explicitly excluded from the claimed invention by way of proviso or negative limitation, such as any specific compounds or method steps, whether implicitly or explicitly defined herein.

[0035] In addition, all ranges given herein include the end of the ranges and also any intermediate range points, whether explicitly stated or not.

[0036] Finally, terms of degree such as "substantially", "about" and "approximately" as used herein mean a reasonable amount of deviation of the modified term such that the end result is not significantly changed. These terms of degree should be construed as including a deviation of at least ±5% of the modified term if this deviation would not negate the meaning of the word it modifies.

[0037] The abbreviation, “e.g.” is derived from the Latin exempli gratia, and is used herein to indicate a non-limiting example. Thus, the abbreviation “e.g.” is synonymous with the term “for example.” The word “or” is intended to include “and” unless the context clearly indicates otherwise.

[0038] The phrase “at least one of” is understood to be one or more. The phrase “at least one of... and...” is understood to mean at least one of the elements listed or a combination thereof, if not explicitly listed. For example, “at least one of A, B, and C” is understood to mean A alone or B alone or C alone or a combination of A and B or a combination of A and C or a combination of B and C or a combination of A, B, and C.

[0039] The specification relates to inhibitors of the nuclear enzyme poly(adenosine 5'-diphospho-ribose) polymerase 1 , which may also be referred to as poly(ADP-ribose) polymerase 1 , PARP-1 , NAD⁺ ADP-ribosyl transferase 1 and poly(ADP-ribose) synthase 1 , and discloses compounds and compositions containing the disclosed compounds. Moreover, the specification discloses a method for treatment of a subject by administration of the disclosed PARP-1 inhibitors. In a particular embodiment, the disclosed PARP-1 inhibitors are used to treat cancer.

[0040] In one aspect the specification relates to a compound of formula (I):

[0041] or a pharmaceutically acceptable salt or pro-drug thereof,

[0042] R¹ and R² independently are H, halogen, a Ci-6-substituent optionally having one or more heteroatoms, OR¹¹, or NR¹²R¹³;

[0043] R¹¹, R¹² and R¹³ are independently selected from H, benzyl, a

Ci-6-substituent optionally having one or more heteroatoms, C(=O)R¹⁴, C(=O)OR¹⁵, or C(=O)NR¹⁶R¹⁷;

[0044] R¹⁴, R¹⁵, R¹⁶, and R¹⁷ are independently selected from H, benzyl, an optionally substituted aryl, an optionally substituted heteroaryl, or a Ci-6-substituent optionally having one or more heteroatoms;

[0045] X is O, S, or NH;

[0046] L¹ is absent or is NH, and when L¹ is absent, the quinazolinone moiety is bonded to L²;

[0047] L² is a Ci-3-chain optionally substituted with one or more methyl or methoxy, and L² being further optionally substituted with one or more heteroatoms;

[0048] L³ is absent or is NH, O, S;

[0049] L⁴ is absent or is - CH2 -;

[0050] L⁵ is absent or is C(=O), C(=S), S(=O) or S(=O)₂;

[0051] wherein when L³ is absent, L² is bonded to L⁴; when L³ and L⁴ are absent, L² is bonded to L⁵; when L³, L⁴ and L⁵ are absent, L² is bonded to R; when L⁴ is absent, L³ is bonded to L⁵; when L⁴ and L⁵ are absent, L³ is bonded to R; and when L⁵ is absent, L⁴ is bonded to R;

[0052] or

[0053] L¹, L², L³, L⁴ and L⁵ together form a three to six membered ring optionally having one or more heteroatoms; and [0054] R is H or a C-i-20-substituent optionally having one or more heteroatoms.

[0055] The compounds of formula (I) as disclosed herein have a quinazolinone moiety bonded to the structure -L¹-L²-L³-L⁴-L⁵-R, as shown in formula (I). The quinazolinone moiety has the structure:

[0056] wherein T represents the partial bond or point of bonding. In the current instance, as shown above, the

represents the partial bond between the quinazolinone moiety, and where it bonds to -L¹-L²-L³-L⁴-L⁵-R. The quinazolinone moiety present in the compound of formula (I) can be further substituted by R¹ and/or

R². Furthermore, the quinazolinone moiety has X, which can be O, S or NH. Nonlimiting example embodiments of the quinazolinone moiety include:

[0057] The term, pharmaceutically acceptable salt, as disclosed herein is not particularly limited and should be known to a skilled worker, or can be determined. There are no particular limitations on the pharmaceutically acceptable salt so long as the compound disclosed herein, and salt are formed, whether inorganic acid salt, inorganic base salt, organic base salt or organic acid salt. For example and without limitation, the salt can be hydrochloric acid salt, sulfuric acid salt, citrate, hydrobromic acid salt, hydroiodic acid salt, nitric acid salt, bisulfate, phosphoric acid salt, isonicotinic acid salt, acetic acid salt, lactic acid salt, salicylic acid salt, tartaric acid salt, pantotenic acid salt, ascorbic acid salt, succinic acid salt, maleic acid salt, fumaric acid salt, gluconic acid salt, saccharinic acid salt, formic acid salt, benzoic acid salt, glutaminic acid salt, methanesulfonic acid salt (also referred to as mesylic acid salt), ethanesulfonic acid salt, benzenesulfonic acid salt, p-toluenesulfonic acid salt, pamoic acid salt (pamoate), sodium salt, potassium salt, calcium salt, magnesium salt, arginine salt, diethanolamine, ethanolamine, ethylenediamine, histidine salt, triethylamine salt and so on.

[0058] The term, pro-drug, as used herein is not particularly limited and should be known to a person of ordinary skill in the art. A prodrug is a compound that is administered in a pharmacologically inactive form which is then converted to an active form through a normal metabolic process, such as hydrolysis of an ester. In other words, a pro-drug is a precursor chemical compound of an active pharmaceutical ingredient. In addition, a pro-drug of the compound disclosed herein can be determined or formed by a person of ordinary skill in the art. There are no particular limitations on the pro-drug of the compounds disclosed herein.

[0059] The term, halogen, as used herein is not particularly limited and should be known or understood by a person of skill in the art. The halogens are elements that form group 17 of the periodic table. In one embodiment, for example and without limitation, the term halogen includes fluorine, chlorine, bromine, or iodine. In a second embodiment, for example and without limitation, the term halogen includes fluorine, chlorine, or bromine.

[0060] The term, Ci-6-substituent, and the like, as used herein is not particularly limited, and should be understood by a person of skill in the art. The term refers to an organic substituent having the number of carbon atoms as noted in the subscript following C (the symbol for carbon). As such, the Ci-6-substituent, and the like, has from one to six carbon atoms. Hence, a Ci-6-substituent, and the like, can have one, two, three, four, five or six carbon atoms. The organic substituent formed by the Ci-6-substituent, and the like, is not particularly limited, and can include linear, branched or cyclic substituents, or a combination thereof. In one embodiment, for example and without limitation, the Ci-6-substituent, and the like, can include alkyl, alkenyl, alkynyls, carbocycle, aryl, a combination thereof and the like. In another embodiment, for example and without limitation, the specification refers to a Ci-20-substituent, and the like, where the organic substituent has from one to twenty carbon atoms.

[0061] The Ci-6-substituent, and the like, as disclosed herein can optionally have one or more heteroatoms. The phrase, optionally having one or more heteroatoms, as used herein is not particularly limited and should be known or understood by a person of skill in the art. A heteroatom refers to any atom other than carbon and hydrogen. In one embodiment, for example and without limitation, the heteroatom is one or more of nitrogen, oxygen, sulphur or a halogen. The presence of the heteroatom can change the functional group of the organic substituent on which the heteroatom is present. For example, and without limitation, when an oxygen atom is present in an alkyl chain, the organic substituent can be an alcohol, an ether, an alkoxide, a ketone, or an aldehyde. The number of heteroatoms present on the organic substituent is not particularly limited and can be varied based on design and application requirements. In one embodiment, for example and without limitation, the organic substituent can have from one, two, three, four, five or six heteroatoms, and where the heteroatoms are the same or different.

[0062] As such, the Ci-6-substituent, and the like, having one or more heteroatoms can include different types of organic substituents, which can include, for example and without limitation, alcohol, ether, aldehyde, ketones, esters, amines, amide, urea, thiols, or thioketones. In addition, when the Ci-6-substituent, and the like, has one or more heteroatoms, the bonding can take place from the carbon atom of the Ci-6-substituent, or from the heteroatom present on the Ci-6-substituent, and the like.

[0063] The term, alkyl, as used herein is not particularly limited and should be known or understood by a person of skill in the art. An alkane as used herein is a saturated hydrocarbon. An alkyl group is an alkane missing one hydrogen. Nonlimiting examples of alkyl include methyl, ethyl, propyl, /so-propyl, butyl, /so-butyl, te/Y-butyl, pentyl, /so-pentyl, etc. The alkyl group may be a straight-chain, a branched-chain or cyclic. The term, alkyl, is intended to embrace all structural isomeric forms of an alkyl group. For example, as used herein, propyl encompasses both n-propyl and /so-propyl; and butyl encompasses n-butyl, sec-butyl, /so-butyl and te/Y-butyl.

[0064] The term, alkenyl, as used herein is not particularly limited and should be known or understood by a person of skill in the art. An alkene as used herein is an unsaturated hydrocarbon having a double bond. An alkenyl group is an alkene missing one hydrogen. Non-limiting examples of alkenyl include ethenyl, propenyl, 1-butenyl, 2-butenyl, pentenyl, /so-pentenyl, /so-propenyl etc. The alkenyl group may be a straight-chain, a branched-chain or cyclic. The term, alkenyl, is intended to embrace all structural isomeric forms of an alkenyl group. For example, as used herein, butenyl encompasses 1 -butenyl and 2-butenyl.

[0065] The term, alkynyl, as used herein is not particularly limited and should be known or understood by a person of skill in the art. An alkyne as used herein is an unsaturated hydrocarbon having a triple bond. An alkynyl group is an alkyne missing one hydrogen. Non-limiting examples of alkynyl include ethynyl, propynyl, 1 -butynyl, 2-butynyl, 1 -pentynyl, 2-pentynyl, 3-methybut-1-yne etc. The alkynyl group may be a straight-chain, a branched-chain or cyclic. The term, alkynyl, is intended to embrace all structural isomeric forms of an alkynyl group. For example, as used herein, pentynyl encompasses 1 -pentynyl, 2-pentynyl, and 3-methybut-1- yne.

[0066] The term, aryl, as used herein is not particularly limited and should be known or understood by a person of skill in the art. An aryl is any functional group or substituent derived from an aromatic ring, usually an aromatic hydrocarbon. In addition, the term, aryl, as used herein, refers to a monocyclic or polycyclic aromatic group. Non-limiting examples of aryl group include phenyl, naphthyl, benzyl, and the like. When the Ci-6-substituent, and the like, have one or more heteroatoms, the aryl group is a heteroaryl, and can include, for example and without limitation, pyridine, furan, pyrazine, imidazole, pyrazole, oxazole, thiophene, and the like.

[0067] In the compound of formula (I), L¹ is absent or is NH, and when L¹ is absent, the quinazolinone moiety is bonded to L². In addition, L² is a Ci-3-chain optionally substituted with one or more methyl or methoxy, and L² being further optionally substituted with one or more heteroatoms. Further, L³ is absent or is NH, O, S; L⁴ is absent or is - CH2 -; and L⁵ is absent or is C(=O), C(=S), S(=O) or S(=O)₂. When L³ is absent, L² is bonded to L⁴; when L³ and L⁴ are absent, L² is bonded to L⁵; when L³, L⁴ and L⁵ are absent, L² is bonded to R; when L⁴ is absent, L³ is bonded to L⁵; when L⁴ and L⁵ are absent, L³ is bonded to R; and when L⁵ is absent, L⁴ is bonded to R. Alternatively, L¹, L², L³, L⁴ and L⁵ together form a three to six membered ring optionally having one or more heteroatoms.

[0068] The term, Ci-3-chain as used herein is not particularly limited, and should be understood by a person of skill in the art. The term refers to an organic substituent having the number of carbon atoms as noted in the subscript following C (the symbol for carbon). As such, the Ci-3-chain has from one to three carbon atoms in the chain length. In addition, like the Ci-6-substituent, the Ci-3-chain can be alkane, alkene, alkyne, cyclic, and the like. In one embodiment, as disclosed herein, the Ci-3-chain is optionally substituted with one or more methyl or methoxy, and L² being further optionally substituted with one or more heteroatoms. The C1-3- chain generally forms a linear chain length, however, when the Ci-3-chain is substituted with one or more methyl or methoxy substituents, L² forms a branched chain. In addition, the Ci-3-chain can optionally have one or more heteroatoms. Non-limiting example of compound of formula (I), having the quinazolinone moiety and L¹, L², L³, L⁴ and L⁴ include:

£1

In the compound of formula (I), R is H or a C-i-20-substituent optionally having one or more heteroatoms. The term, Ci-20-substituent optionally having one or more heteroatoms, as used herein is not particularly limited and should be understood by a person of skill in the art. As noted above, the term refers to an organic substituent having the number of carbon atoms as noted in the subscript following C (the symbol for carbon). As such, the C-i-20-substituent, and the like, has from one to twenty carbon atoms. The organic substituent formed by the Ci-20-substituent, and the like, is not particularly limited, and can include linear, branched or cyclic substituents, or a combination thereof. In one embodiment, for example and without limitation, the C1- 20-substituent, and the like, can include alkyl, alkenyl, alkynyls, carbocycle, aryl, a combination thereof and the like. The term C-i-20-substituent is like the C1-6- substituent, with the difference being that it can contain up to 20 carbon atoms, optionally having one or more heteroatoms. Furthermore, like the Ci-6-substituent, in the Ci-20-substituent, the heteroatoms can branch off the Ci-20-substituent or be part of a chain in the Ci-20-substituent. Non-limiting examples of R present as part of the compound of formula (I) include:

[0070] In one embodiment, for example and without limitation, the specification relates to a compound of formula (I), as disclosed herein, or a pharmaceutically acceptable salt or pro-drug thereof, wherein

[0071] R¹ and R² independently are H, halogen, or OR¹¹;

[0072] R¹¹ is H, benzyl, or a Ci-6-substituent optionally having one or more heteroatoms; and

[0073] X is O, or S.

[0074] As should be recognized by a person of skill in the art, the compound of formula (I) can include enantiomers and diastereomers. The specification encompasses such enantiomers and diastereomers.

[0075] The term, enantiomer, as used herein is not particularly limited and should be known or understood by a person of skill in the art. In organic chemistry, an enantiomer is a type of stereoisomer. Enantiomers, also known as optical isomers, are two stereoisomers that are related to each other by a reflection. Enantiomers are a pair of molecules that exist in two forms that are mirror images of one another but cannot be superimposed one upon the other. Enantiomers are in every other respect chemically identical.

[0076] The term, diastereomer, as used herein is not particularly limited and should be known or understood by a person of skill in the art. In organic chemistry, diastereomers are a type of stereoisomer. Diastereomers are defined as non-mirror image, non-identical stereoisomers. Hence, they occur when two or more stereoisomers of a compound have different configurations at one or more of the equivalent stereocenters and are not mirror images of each other. [0077] In a second aspect, the specification relates to a method for treating diseases mediated by PARP-1 protein comprising administering to a mammal in need thereof an effective amount of the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, or combination thereof, as disclosed herein.

[0078] In a third aspect, the specification relates to an in vitro method for inhibiting PARP-1 protein activity, comprising contacting the said protein with an effective amount of the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, or combination thereof, as disclosed herein.

[0079] In a fourth aspect, the specification relates to a pharmaceutical composition comprising a therapeutically effective amount of the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, or combination thereof, as disclosed herein, and at least one pharmaceutically acceptable excipient, carrier or diluent.

[0080] In a fifth aspect, the specification relates to a product comprising the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, or combination thereof, as disclosed herein, and one or more chemotherapeutic agents, as a combined preparation for simultaneous, separate or sequential use in anticancer therapy.

[0081] In a fifth aspect, the specification relates to use of the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, or combination thereof, as disclosed herein, in the manufacture of a medicament for treatment of cancer.

[0082] In a sixth aspect, the specification relates to use of the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, or combination thereof, as disclosed herein, for treating cancer.

[0083] The term, pharmaceutical composition, as used herein is not particularly limited and should be known or understood by a person of skill in the art. Pharmaceutical composition means one or more active ingredients, and one or more inert ingredients that make up the carrier, as well as any product which results, directly or indirectly, from combination, complexation or aggregation of any two or more of the ingredients, or from dissociation of one or more of the ingredients, or from other types of reactions or interactions of one or more of the ingredients. Accordingly, the pharmaceutical compositions of the present disclosure can encompass any composition made by admixing at least one compound of the present disclosure and a pharmaceutically acceptable carrier.

[0084] The term, therapeutically effective amount or effective amount, as used herein is not particularly limited and should be understood by a person of skill in the art. The therapeutically effective amount of a compound described herein, a pharmaceutically acceptable salt, tautomer, prodrug, or deuterated analog thereof means an amount sufficient to effect treatment when administered to a subject, to provide a therapeutic benefit such as amelioration of symptoms or slowing of disease progression. For example, a therapeutically effective amount may be an amount sufficient to decrease a symptom of a disease or condition responsive to PARP-1 inhibitors. The therapeutically effective amount may vary depending on the subject, and disease or condition being treated, the weight and age of the subject, the severity of the disease or condition, and the manner of administering, which can be determined by a person of skill in the art.

[0085] The term, pharmaceutically acceptable carrier, as used herein is not particularly limited and should be known or understood by a person of skill in the art. Pharmaceutically acceptable carrier can include one or more excipients or agents such as solvents, diluents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like that are not deleterious to the disclosed compound or use thereof. The use of such carriers and agents to prepare compositions of pharmaceutically active substances is well known in the art (see, e.g., Remington’s Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, PA 17th Ed. (1985); and Modern Pharmaceutics, Marcel Dekker, Inc. 3rd Ed. (G.S. Banker s C.T. Rhodes, Eds., both incorporated herein by reference)

[0086] Non-limiting examples of pharmaceutically acceptable carriers or diluents include water, sodium chloride (NaCI), normal saline solutions, lactated Ringer's, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (such as Ringer's solution), alcohols, oils, gelatins, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethycellulose, polyvinyl pyrrolidine, and colors, and the like. Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and/or aromatic substances and the like that do not deleteriously react with or interfere with the activity of the compounds provided herein. One of ordinary skill in the art will recognize that other pharmaceutical excipients are suitable for use with disclosed compounds.

[0087] Conventional procedures and ingredients for the selection and preparation of suitable pharmaceutical compositions are described, for example, in Remington's Pharmaceutical Sciences (2003 -20th edition) and in The United States Pharmacopeia: The National Formulary (USP 24 NF19) published in 1999 (both incorporated herein by reference). The one or more carriers are "acceptable" in the sense of being compatible with the other ingredients of the pharmaceutical composition and not deleterious to the recipient thereof.

[0088] In a further aspect, the specification provides methods to inhibit, limit and/or control the in vitro and/or in vivo polymerase activity of poly(ADP-ribose) polymerase (PARP) in solutions cells, tissues, organs or organ systems. In one embodiment, the present specification provides methods of limiting or inhibiting PARP activity in a mammal, such as a human, for example and without limitation, either through local or systemic administration.

[0089] In another further aspect, the specification provides a chemosensitization method for treating cancer, the method containing the step of contacting the cancer cells with a cytotoxicity-potentiating quinazolinone compound of Formula (I), a pharmaceutically acceptable salt, or a pro-drug thereof, and further contacting the tumor or cancer cells with an anticancer agent.

[0090] In a still another aspect, the specification provides a chemosensitization method wherein a first dose of at least one compound of Formula (I), a pharmaceutically acceptable salt, or a pro-drug thereof, is administered singly or repeatedly to a patient in need thereof, and wherein subsequently a second dose of at least one chemotherapeutic agent is administered singly or repeatedly to said patient after a time period to provide an effective amount of chemosensitization.

[0091] An aspect of the present specification provides a pharmaceutical formulation comprising the compound of Formula (I) in a form, for example and without limitation, pharmaceutically acceptable free base, salt, hydrate, ester, solvate, stereoisomer, and mixtures thereof. According to a further aspect, the pharmaceutical formulation further comprises a pharmaceutically acceptable carrier or diluent, and, optionally, a chemotherapeutic agent. The following embodiments are for illustrative purposes only and are not intended to limit in any way the scope of the present specification. In one embodiment, a pharmaceutical formulation, as disclosed herein, comprises a compound, as disclosed herein, in a pharmaceutically acceptable carrier. In another embodiment, a pharmaceutical formulation, as disclosed herein, comprises a pharmaceutically acceptable salt of a compound, as disclosed herein, in a pharmaceutically acceptable carrier. In another embodiment, a pharmaceutical formulation, as disclosed herein, comprises a compound, as disclosed herein, and one or more chemotherapeutic agents in a pharmaceutically acceptable carrier. In another embodiment, a pharmaceutical formulation, as disclosed herein, comprises a pharmaceutically acceptable salt of a compound, as disclosed herein, and one or more chemotherapeutic agents in a pharmaceutically acceptable carrier. Non-limiting examples of such chemotherapeutic agents are noted below.

[0092] In another further aspect in accordance with the specification, the chemosensitizing compound and the chemotherapeutic agent can be administered essentially simultaneously. In another further aspect in accordance with the specification, the chemosensitizing compound and the chemotherapeutic agent can be administered essentially sequentially.

[0093] In a particular embodiment, the chemotherapeutic agent is, for example and without limitation, temozolomide, adriamycin, camptothecin, carboplatin, cisplatin, daunorubicin, docetaxel, doxorubicin, interferon-alpha, interferon-beta, interferongamma, interleukin 2, irinotecan, paclitaxel, a taxoid, dactinomycin, danorubicin, 4'- deoxydoxorubicin, bleomycin, pilcamycin, mitomycin, neomycin and gentamycin, etoposide, 4-OH cyclophosphamide, a platinum coordination complex, topotecan, therapeutically effective analog or derivative of the same, or a mixture thereof. In a particular embodiment, the chemotherapeutic agent is temozolomide.

[0094] In another aspect, the present specification provides a method of treating the effect of cancer and/or to radiosensitize cancer cells to render the cancer cells more susceptible to radiation therapy and thereby to prevent the tumor cells from recovering from potentially lethal damage of DNA after radiation therapy, the method containing the step of administering to a subject an effective amount of a compound of Formula (I), a pharmaceutically acceptable salt, ora pro-drug thereof. In a particular embodiment, the method is directed to radiosensitizing cancer cells rendering the cancer cells more susceptible to radiation therapy than non-tumor cells.

[0095] In still further aspect, the specification provides a method of treatment of cancer in a subject in need thereof containing the step of administering to the subject a therapeutically effective amount of a compound of Formula (I), a pharmaceutically acceptable salt, or a pro-drug thereof, wherein the cancer cells have a defect in repair of double-stranded DNA scission. In one embodiment, the defect in repair of doublestranded DNA scission is a defect in homologous recombination. In another embodiment, the cancer cells have a phenotype selected from, for example and without limitation, a BRCA-1 defect, a BRCA-2 defect, a BRCA-1 and BRCA-2 defect, or Fanconi anemia.

[0096] In one embodiment, the present specification provides methods of treating BRCA1/2-associated breast cancer containing the step of administering a compound of Formula (I), a pharmaceutically acceptable salt, or a pro-drug thereof.

[0097] In accordance with another aspect, the specification discloses a compound for use in the chemosensitization method disclosed herein, the radiosensitization method disclosed herein, or the treatment of cancer wherein the cancer cells have a defect in repair of double-stranded DNA scission method, where the compound is selected from Formula (I), a pharmaceutically acceptable salt, or a pro-drug thereof.

[0098] The process for synthesis of the compound of formula (I) is not particularly limited, and is further disclosed in the Examples below. In addition, methodology similar to that disclosed in PCT patent publication number WO 2018/125961 (incorporated herein by reference), along with common general knowledge, can be used for synthesis of the compound of formula (I), or a pharmaceutically acceptable salt or pro-drug thereof. In one embodiment, the compounds disclosed in the current specification exclude compounds disclosed in WO 2018/125961.

EXAMPLES

[0099] The above disclosure generally describes the present invention. A more complete understanding can be obtained by reference to the following specific Examples. These Examples are described solely for purposes of illustration and are not intended to limit the scope of the invention. Changes in form and substitution of equivalents are contemplated as circumstances may suggest or render expedient. Without further description, it is believed that one of ordinary skill in the art can, using the preceding description and the following illustrative examples, make and utilize the constructs of the present invention and practice the claimed methods. The following working examples therefore, specifically point out the typical aspects of the present invention and are not to be construed as limiting in any way in the remainder of the disclosure. Although specific terms have been employed herein, such terms are intended in a descriptive sense and not for purposes of limitation.

[00100] A. Aldehyde intermediate (10) synthesis

Scheme 1 : Synthesis of advanced intermediate Aldehyde 10

[00101 ] 4-(Benzyloxy)-1 -(trityloxy)-2-butanol (2)

[00102] 4-(Benzyloxy)-1 -(trityloxy )-2-butanol (2) was synthesized from 2-(2,2-

Dimethyl-1 ,3-dioxolan-4-yl)ethanol (1), following the synthetic route and procedures described in Carbohydrate Research (1996), vol. 294, p 65-94.

[00103] ((4-(Benzyloxy)-2-methoxybutoxy)methanetriyl)tribenzene (3)

[00104] 4-(Benzyloxy)-1 -(trityloxy )-2-butanol (2) (1 equiv.) was dissolved in dimethyl formamide (DMF) and cooled to 0°C. Sodium hydride (1.2 equiv.) was then added in portions with stirring. The mixture was stirred for a further 6 h at ambient temperature. After quenching the excess NaH, the solvent was removed, and the residue was dissolved in dichloromethane (DCM). The organic layer was washed using water (x3), dried (MgSO4), filtered, and concentrated to give title compound 3 as a pure oily product (96%) which was used as such in the next step. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.63-7.25 (m, 20H), 4.56 (s, 2H), 3.64-3.52 (m, 3H), 3.48 (s, 3H), 3.31-3.18 (m, 2H), 2.02-1.88 (m, 2H). ES-MS: 475 [M+Na]+.

[00105] 4-(Benzyloxy)-2-methoxybutan-1-ol (4)

[00106] The trityl-protected compound 3 was dissolved in a minimum amount of DCM and diluted using MeOH. Amberlyst-15 was added, and the mixture was stirred at ambient temperature for 5 h. The solvents were removed followed by purification using column chromatography (Hexanes/EtOAc) to give the title compound 4 as a colorless oily product (87%). ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.38-7.25 (m, 5H), 4.60 (s, 2H), 3.75-3.68 (m, 1 H), 3.63-3.38 (m, 7H), 2.81 (s, 1 H), 1.88-1.73 (m, 2H). ES-MS: 233 [M+Na]+.

[00107] 4-(Benzyloxy)-2-methoxybutanoic acid (5)

[00108] The alcohol 4 (1 equiv.) was dissolved in acetone and the solution was cooled to 0°C. A Jones reagent solution (1.5 equiv.) was added slowly with stirring. The mixture was stirred for a further 6h at ambient temperature and concentrated. The residue was dissolved in EtOAc, and this organic layer was washed using water (x4). The organic layer was extracted using 10 % NaOH (x4) and discarded. The aqueous extracts were combined, cooled to 0°C and acidified to pH ~ 3 or until cloudy. The pure acid 5 (74%) was then extracted from this acidic solution using DCM. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.38-7.29 (m, 5H), 4.52-4.48 (m, 2H), 4.06-3.94 (m, 1 H), 3.66-3.61 (m, 2H), 3.38 (s, 3H), 2.24-1.88 (m, 2H). ES-MS: 247 [M+Na]+.

[00109] 2-(4-(Benzyloxy)-2-methoxybutanamido)-3-methoxybenzamide (7)

[00110] Carboxylic acid 5 (1 equiv.) and EtaN (2 equiv.) were dissolved in tetrahydrofuran (THF) and the mixture was stirred at 0°C for 10 min. Methanesulfonyl chloride (1.5 equiv.) was added and the mixture was stirred at ambient temperature for up to 1 h. 2-Amino-3-methoxybenzamide (6) in THF was then added and the mixture was stirred at ambient temperature overnight. The reaction mixture was concentrated followed by purification using column chromatography (EtOAc) to give the product 7 (71 %) as a colorless solid. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.51-8.38 (br s, 1 H), 7.38-7.25 (m, 6H), 7.23-7.13 (m, 1 H), 7.06-6.88 (m, 1 H), 6.50- 6.13 (br s, 1 H), 5.63-5.37 (br s, 1 H), 4.63-4.47 (m, 2H), 4.00-3.89 (m, 1 H), 3.86 (s, 3H), 3.74-3.63 (m, 2H), 3.56 (s, 3H), 2.26-2.13 (m, 1 H), 2.12-1.97 (m, 1 H). ES-MS: 395 [M+Na]+.

[00111 ] 2-(3-(Benzyloxy)-1 -methoxypropyl)-8-methoxyquinazolin-4(3/7)- one (8)

[00112] Compound 7 was dissolved in ethanol and a 1 M solution of NaOH was added until strongly basic. The mixture was stirred at ambient temperature for 1 h and the pH was adjusted to ~ 8 to precipitate the neutral colorless solid product 8 (81 %). ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 9.65-9.40 (br s, 1 H), 7.94-7.81 (m, 1 H), 7.48-7.38 (m, 1 H), 7.37-7.14 (m, 6H), 4.60-4.38 (m, 3H), 4.02 (s, 3H), 3.69-3.62 (m, 2H), 3.38 (s, 3H), 2.38-2.01 (m, 2H). ES-MS: 377 [M+Na]+.

[00113] 2-(3-Hydroxy-1 -methoxypropyl)-8-methoxyquinazolin-4(3/7)-one (9)

[00114] The compound 8 was dissolved in a minimum amount of DCM then diluted using THF and MeOH. A catalytic amount of Pd/C was added, and the reaction vessel was evacuated and back filled with hydrogen gas (x2). The mixture stirred under hydrogen atmosphere for 5 h. The mixture was filtered using Celite and the Celite cake was washed using MeOH. The combined filtrates were concentrated to give colorless solid product 9 (92%). ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.77- 7.72 (m, 1 H), 7.51-7.39 (m, 1 H), 7.38-7.20 (m, 1 H), 4.47-4.37 (m, 1 H), 4.01 (s, 3H), 3.72-3.63 (m, 2H), 3.37 (s, 3H), 2.20-1.88 (m, 2H). ES-MS: 287 [M+Na]+.

[00115] 3-Methoxy-3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanal (10)

[00116] Dess-Martin periodinane (1.5-2 equiv.) was added at 0°C to a solution of alcohol 9 (1 equiv.) and NaHCOa (2-4 equiv.) in anhydrous DCM. The solution was stirred for 10 min and then allowed to warm to ambient temperature where it was stirred for a further two to three hours. A 1 :1 :1 mixture of saturated aqueous sodium thiosulfate solution, saturated aqueous sodium bicarbonate solution, and water was added slowly. The resulting biphasic mixture was stirred vigorously for 1 h resulting in two clear layers. The layers were separated, and the aqueous layer was extracted three times with DCM. The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The resulting aldehyde was subjected to General Procedure A without further purification.

[00117] B. Preparation of PARP Inhibitors Library following Synthetic

Route I

Scheme 2: Synthetic Route I

[00118] General Procedure A

[00119] A solution of 3-methoxy-3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanal 10 (1 equiv.) and amine (1.3 equiv.) in 1 ,2-dichloroethane was stirred for 30 min at room temperature, and subsequently, sodium triacetoxyborohydride (2 equiv.) was added. The mixture was stirred overnight at ambient temperature. The solvent was removed in vacuo and the residue was purified using chromatography (silica) using a gradient of MeOH in DCM. Purification was also achieved using semipreparative HPLC (C column), eluting with a gradient of MeOH vs 0.1 % Formic Acid in water, yielding Amine 11 and/or Amide 12.

[00120] General Procedure B

[00121] The corresponding dimethoxy-protected quinazolinone species (Amine

11 or Amide 12) (1 equiv.) was dissolved in anhydrous DCM or 1 ,2-dichloroethane and excess BBrs solution (1.0 M, 6 to 10 equiv.) was added under nitrogen. The mixture was heated to reflux and for a duration of 6 to 18 h. The solvent was removed, and purification was achieved using semi-preparative HPLC (Cis column), eluting with a gradient of MeOH vs 0.1 % Formic Acid in water, yielding di-hydroxy compound Amine 13 or Amide 15, from Amine 11 or Amide 12 respectively.

[00122] General Procedure C

[00123] The corresponding methoxyquinazolin-4(3/7)-one protected species

(Amine 11 or Amide 12) (1 equiv.) was dissolved in anhydrous DCM and excess BBra solution (1.0 M, 2 to 4 equiv.) was added under nitrogen. The mixture was stirred at room temperature overnight. The solvent was removed, and purification was achieved using semi-preparative HPLC (Cis column), eluting with a gradient of MeOH vs 0.1 % Formic Acid in water, yielding mono-hydroxy compound Amine 14 or Amide 16, from Amine 11 or Amide 12 respectively.

[00124] 3-Methoxy-3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2-yl)-A/-(2- morpholinoethyl)propenamide (101 )

[00125] Prepared using General Procedure A, using compound 10 and 2- Morpholinoethanamine. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 10.61-9.73 (br s, 1 H), 8.19-7.69 (br s, 1 H), 7.62-6.92 (m, 2H), 6.84-6.17 (br s, 1 H), 5.10-4.46 (br s, 1 H), 4.41-3.11 (m, 12H), 3.06-1.99 (m, 9H). ES-MS: 391 [M+H]+.

[00126] (2S)-Methyl 1 -(3-methoxy-3-(8-methoxy-4-oxo-3,4- dihydroquinazolin-2-yl)propyl)pyrrolidine-2-carboxylate (102)

[00127] Prepared using General Procedure A, using compound 10 and L-Proline methyl ester. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.82-7.71 (m, 1 H), 7.52-7.42 (m, 1 H), 7.40-7.32 (m, 1 H), 4.41-4.28 (m, 1 H), 4.00 (s, 3H), 3.78-3.58 (m, 3H), 3.39 (s, 3H), 3.29-2.79 (m, 3H), 2.67-2.28 (m, 2H), 2.16-1.92 (m, 3H), 1.90-1.62 (m, 3H). ES-MS: 376 [M+H]+.

[00128] (2S)-Methyl 1 -(3-methoxy-3-(8-methoxy-4-oxo-3,4- dihydroquinazolin-2-yl)propanoyl)pyrrolidine-2-carboxylate (103)

[00129] Prepared using General Procedure A, using compound 10 and L-Proline methyl ester. ¹H NMR (300 MHz, Methanol-cM): 6 (ppm) 7.82-7.69 (d, 1 H), 7.51-7.39 (t, 1 H), 7.38-7.29 (d, 1 H), 4.80-4.68 (m, 1 H), 4.49-4.37 (m, 1 H), 3.99 (s, 3H), 3.79- 3.61 (m, 4H), 3.59-3.35 (m, 3H), 3.15-2.89 (m, 2H), 2.39-2.14 (m, 1 H), 2.12-1.71 (m, 3H). ES-MS: 390 [M+H]+.

[00130] (2S)-1 -(3-Hydroxy-3-(8-hydroxy-4-oxo-3,4-dihydroquinazolin-2- yl)propyl)pyrrolidine-2 -carboxylic acid (104)

[00131] Prepared using General Procedure B, from compound 102. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.75 (d, 1 H), 7.58 (dt, 1 H), 7.43 (d, 1 H), 5.38-5.17 (m, 1 H), 4.55 (t, 1 H), 4.09-3.70 (m, 2H), 3.69-3.51 (m, 1 H), 3.49-3.32 (m, 1 H), 2.81- 1.93 (m, 6H). ES-MS: 334 [M+H]+.

[00132] 8-Methoxy-2-(1 -methoxy-3-(4-(2-methoxyethyl)piperazin-1 - yl)propyl)quinazolin-4(3/7)-one (105)

[00133] Prepared using General Procedure A, using compound 10 and 1-(2- Methoxyethyl)piperazine. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.67-7.53 (m, 1 H), 7.37-7.11 (m, 2H), 4.24-4.08 (br s, 1 H), 3.83 (s, 3H), 3.24-3.05 (m, 6H), 2.71- 2.02 (m, 14H), 1.92-1.80 (br s, 2H). ES-MS: 391 [M+H]+.

[00134] Preparation of 8-Hydroxy-2-(1 -hydroxy-3-(4-(2- methoxyethyl)piperazin-1 -yl)propyl)quinazolin-4(3/7)-one (106), 8-Hydroxy-2-(1 - hydroxy-3-(4-(2-hydroxyethyl)piperazin-1 -yl)propyl)quinazolin-4(3/7)-one (107), 2-(1-Bromo-3-(4-(2-methoxyethyl)piperazin-1-yl)propyl)-8-hydroxyquinazolin- 4(3/7)-one (108), and 2-(1-Bromo-3-(4-(2-hydroxyethyl)piperazin-1-yl)propyl)-8- hydroxyquinazolin-4(3/7)-one (109)

Mixture of 4 compounds: Purification / Separation by Semi-preparative HPLC

[00135] 8-Hydroxy-2-(1 -hydroxy-3-(4-(2-methoxyethyl)piperazin-1 - yl)propyl)quinazolin-4(3/7)-one (106)

[00136] Prepared using General Procedure B, from compound 105, obtained after purification by semi-preparative HPLC. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.80 (d, 1 H), 7.51 (t, 1 H), 7.43 (d, 1 H), 4.85 -4.74 (m, 1 H), 4.04 (s, 3H), 3.87- 3.77 (m, 2H), 3.24-2.91 (m, 10H), 2.32-2.08 (m, 2H), 1.44-1.27 (m, 2H). ES-MS: 363 [M+H]+.

[00137] 8-Hydroxy-2-(1 -hydroxy-3-(4-(2-hydroxyethyl)piperazin-1 - yl)propyl)quinazolin-4(3/7)-one (107)

[00138] Prepared using General Procedure B, from compound 105, obtained after purification by semi-preparative HPLC. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.69 (d, 1 H), 7.40 (t, 1 H), 7.28 (d, 1 H), 4.83 (t, 1 H), 3.84 (t, 2H), 3.66-3.46 (br s, 1 H), 3.33 (s, 3H), 3.27-2.95 (m, 8H), 2.42-2.13 (m, 2H). ES-MS: 349 [M+H]+.

[00139] 2-(1 -Bromo-3-(4-(2-methoxyethyl)piperazin-1 -yl)propyl)-8- hydroxyquinazolin-4(3/7)-one (108)

[00140] Prepared using General Procedure B, from compound 105, obtained after purification by semi-preparative HPLC. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.80 (d, 1 H), 7.58-7.36 (m, 2H), 4.85-4.77 (m, 1 H), 4.04 (s, 3H), 3.89-3.78 (m, 1 H), 3.77-3.62 (m, 1 H), 3.61-3.48 (m, 1 H), 3.25-3.02 (m, 4H), 3.01-2.75 (m, 3H), 2.51-2.02 (m, 3H). ES-MS: 425, 427 [M+H]+.

[00141 ] 2-( 1 -Bromo-3-(4-(2-hydroxyethyl)piperazin-1 -yl)propyl)-8- hydroxyquinazolin-4(3/7)-one (109)

[00142] Prepared using General Procedure B, from compound 105, obtained after purification by semi-preparative HPLC. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.75-7.65 (m, 1 H), 7.47-7.35 (m, 1 H), 7.34-7.24 (m, 1 H), 4.87-4.77 (m, 1 H), 3.98-3.86 (m, 1 H), 3.70-3.49 (m, 6H), 3.48-3.34 (m, 5H), 3.13-2.93 (m, 2H), 2.56- 2.19 (m, 2H). ES-MS: 411-413 [M+H]+.

[00143] 3-Methoxy-3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2-yl)-N-(2-(1 - methylpyrrolidin-2-yl)ethyl)propenamide (110)

[00144] Prepared using General Procedure A, using compound 10 and 2-(1- Methylpyrrolidin-2-yl)ethanamine. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.78 (d, 1 H), 7.48 (t, 1 H), 7.39 (d, 1 H), 4.69 (t, 1 H), 4.01 (s, 3H), 3.43 (s, 3H), 3.24 (t, 2H), 3.14-2.96 (m, 1 H), 2.89-2.68 (m, 2H), 2.39-1 .84 (m, 10 H), 1.83-1.64 (m, 2H), 1.62- 1.29 (m, 2H). ES-MS: 389 [M+H]+.

[00145] 8-Methoxy-2-(1 -methoxy-3-(2-(1 -methylpyrrolidin-2- yl)ethylamino)propyl)quinazolin-4(3/7)-one (111 )

[00146] Prepared using General Procedure A, using compound 10 and 2-(1- Methylpyrrolidin-2-yl)ethanamine. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.77 (d, 1 H), 7.45 (t, 1 H), 7.34 (d, 1 H), 4.36 (t, 1 H), 4.01 (s, 3H), 3.38 (s, 3H), 3.12-2.96 (m, 1 H), 2.95-2.55 (m, 4H), 2.42-1.86 (m, 10H), 1.84-1.64 (m, 2H), 1.59-1.35 (m, 2H). ES-MS: 375 [M+H]+.

[00147] 3-Hydroxy-3-(8-hydroxy-4-oxo-3,4-dihydroquinazolin-2-yl)-N-(2-(1 - methylpyrrolidin-2-yl)ethyl)propenamide (112)

[00148] Prepared using General Procedure B, from compound 110. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.67 (d, 1 H), 7.38 (t, 1 H), 7.25 (d, 1 H), 5.04 (t, 1 H), 3.81-3.53 (m, 2H), 3.27-3.15 (m, 1 H), 3.14-2.99 (m, 1 H), 2.98-2.66 (m, 5H), 2.52- 2.26 (m, 1 H), 2.24-1.90 (m, 4H), 1.85-1.49 (m, 2H). ES-MS: 361 [M+H]+.

[00149] 8-Hydroxy-2-(1 -hydroxy-3-(2-(1 -methylpyrrolidin-2- yl)ethylamino)propyl)quinazolin-4(3/7)-one (113)

[00150] Prepared using General Procedure B, from compound 111. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.69 (d, 1 H), 7.39 (t, 1 H), 7.28 (d, 1 H), 4.85-4.76 (m, 1 H), 3.85-3.60 (br, 1 H), 2.96 (s, 3H), 2.61-1.71 (m, 8H). ES-MS: 346 [M+H]+.

[00151 ] 6-{4-[3-Methoxy-3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propyl]piperazin-1 -yl}pyridine-3-carbonitrile (114)

[00152] Prepared using General Procedure A, from compound 10 and (6- (Piperazin-1-yl)pyridine-3-carbonitrile). Purification by column chromatography (DCM/MeOH 90:10) yielded the title compound as a colorless solid. ¹H NMR (300 MHz, Chloroform-d): 6 (ppm) 11.08 (s, 1 H), 8.38 (s, 1 H), 7.86 (d, 1 H), 7.60 (dd, 1 H), 7.46 (dd, 1 H), 7.22 (dd, 1 H), 6.58 (d, 1 H), 4.49 (dd, 1 H), 4.09 (s, 3H), 3.62 (m, 4H), 3.40 (s, 3H), 2.74 (m, 1 H), 2.54 (m, 4H), 2.48 (m, 1 H), 2.16 (m, 2H). ES-MS: 435 [M+H]+.

[00153] 6-{4-[3-Methoxy-3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl]piperazin-1 -yl}pyridine-3-carbonitrile (115)

[00154] Prepared using General A, from compound 10 and (6-(Piperazin-1- yl)pyridine-3-carbonitrile). Purification by column chromatography (CH2Cl2/MeOH 90:10) yielded the title compound as a colorless solid. ¹H NMR (300 MHz, Chloroformed): 5 (ppm) 10.05 (s, 1 H), 8.38 (s, 1 H), 7.86 (d, 1 H), 7.62 (dd, 1 H), 7.46 (dd, 1 H), 7.22 (dd, 1 H), 6.56 (d, 1 H), 4.89 (dd, 1 H), 3.98 (s, 3H), 3.72 (m, 4H), 3.65 (m, 2H), 3.59 (m, 2H), 3.52 (s, 3H), 2.98 (m, 2H). ES-MS: 449 [M+H]+. [00155] 6-{4-[3-Hydroxy-3-(8-hydroxy-4-oxo-3,4-dihydroquinazolin-2- yl)propyl]piperazin-1 -yl}pyridine-3-carbonitrile (116)

[00156] Prepared using General Procedure B from compound 114. Purification by semi-preparative HPLC yielded the title compound as a colorless solid. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.88 (s, 1 H), 8.24 (d, 1 H), 8.08 (d, 1 H), 7.74 (dd, 1 H), 7.62 (d, 1 H), 7.32 (d, 1 H), 5.22 (dd, 1 H), 4.46 (m, 4H), 3.92 (m, 6H), 2.85 (m, 2H). ES-MS: 407 [M+H]+.

[00157] 6-(4-(3-(8-hydroxy-4-oxo-3,4-dihydroquinazolin-2-yl)-3- methoxypropyl)piperazin-1 -yl)nicotinonitrile (117)

[00158] Prepared using General Procedure C from compound 114. Purification using semi-preparative HPLC yielded the title compound as a colorless solid. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.41 (s, 1 H), 7.68-7.85 (m, 2H), 7.36-7.53 (m, 2H), 6.79-6.88 (d, 1 H), 4.75-4.87 (m, 1 H), 4.04 (s, 3H), 3.53-3.74 (m, 4H), 2.81-2.95 (m, 1 H), 2.61-2.79 (m, 5H), 2.12-2.26 (m, 1 H). ES-MS: 419.8 [M-H]-.

[00159] 6-{4-[3-Hydroxy-3-(8-hydroxy-4-oxo-3,4-dihydroquinazolin-2-yl) propanoyl]piperazin-1 -yl}pyridine-3-carbonitrile (118)

[00160] By General Procedure B from compound 115. Purification using semipreparative HPLC yielded the title compound as a colorless solid. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.59 (s, 1 H), 8.24 (d, 1 H), 7.68 (d, 1 H), 7.44 (dd, 1 H), 7.29 (d, 1 H), 7.12 (d, 1 H), 5.18 (dd, 1 H), 3.82 (m, 8H), 3.18 (m, 2H). ES-MS: 422 [M+H]+.

[00161 ] 6-{4-[3-(8-Hydroxy-4-oxo-3,4-dihydroquinazolin-2-yl)-3- methoxypropanoyl] piperazin-1 -yl}pyridine-3-carbonitrile (119)

[00162] Prepared using General Procedure C from compound 115. Purification using semi-preparative HPLC yielded the title compound as a colorless solid. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.42 (s, 1 H), 7.78 (d, 1 H), 7.71 (d, 1 H), 7.40 (dd, 1 H), 7.25 (d, 1 H), 6.89 (d, 1 H), 5.14 (dd, 1 H), 3.82 (m, 11 H), 3.19 (m, 2H). ES-MS: 435 [M+H]+.

[00163] 2-[3-(4-Hydroxy-4-methylpiperidin-1 -yl)-1 -methoxypropyl]-8- methoxy-3,4-dihydroquinazolin-4-one (120)

[00164] Prepared using General Procedure A, from compound 10 and 4- Hydroxy-4-methylpiperidine. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.83 (d, 1 H), 7.41 (dd, 1 H), 7.18 (d, 1 H), 4.42 (dd, 1 H), 4.02 (s, 3H), 3.40 (s, 3H), 2.99 (m, 2H), 2.78 (m, 2H), 2.55 (m, 2H), 2.12 (m, 1 H), 1.84 (m, 2H), 1.61 (m, 2H), 1.22 (m, 5H). ES-MS: 362 [M+H]+.

[00165] 8-Methoxy-2-{1 -methoxy-3-[({4-[(4-methylpiperazin-1 - yl)methyl]phenyl} methyl)amino]propyl}-3,4-dihydroquinazolin-4-one (121 )

[00166] Prepared using General Procedure A, from compound 10 and 4-[(4- Methyl-1-piperazinyl)methyl]benzylamine. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.82 (d, 1 H), 7.44 (dd, 1 H), 7.42 (d, 2H), 7.11 (d, 2H), 7.09 (d, 2H), 4.42 (dd, 1 H), 4.21 (m, 2H), 3.98 (s, 3H), 3.82 (s, 2H), 3.48 (m, 5H), 3.11 (m, 8H), 2.82 (s, 3H), 2.32 (m, 2H). ES-MS: 466 [M+H]+.

[00167] 3-Methoxy-3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2-yl)-N-({4-

[(4-methylpiperazin-1-yl)methyl]phenyl}methyl)propenamide (122)

[00168] Prepared using General Procedure A, from compound 10 and 4-[(4- Methyl-1-piperazinyl)methyl]benzylamine. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.75 (d, 1 H), 7.38 (dd, 1 H), 7.21 (m, 6H), 4.73 (dd, 1 H), 4.41 (m, 2H), 3.82 (s, 3H), 3.42 (m, 5H), 2.84 (m, 1 H), 2.78 (m, 1 H), 2.54 (m, 8H), 2.32 (s, 3H). ES-MS: 480 [M+H]+.

[00169] 3-(8-Hydroxy-4-oxo-3,4-dihydroquinazolin-2-yl)-3-methoxy-N-({4-

[(4-methylpiperazin-1-yl)methyl]phenyl}methyl)propenamide (123)

[00170] Prepared using General Procedure C from compound 122. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.66 (d, 1 H), 7.38 (dd, 1 H), 7.27 (d, 2H), 7.18 (m, 3H), 5.06 (dd, 1 H), 4.38 (m, 2H), 3.98 (m, 3H), 3.62 (s, 3H), 3.44 (m, 2H), 3.12 (s, 3H), 2.83 (m, 5H), 2.62 (m, 2H). ES-MS: 466 [M+H]+.

[00171] 3-Hydroxy-3-(8-hydroxy-4-oxo-3,4-dihydroquinazolin-2-yl)-N-({4-

[(4-methylpiperazin-1-yl)methyl]phenyl}methyl)propenamide (124)

[00172] Prepared using General Procedure B from compound 122. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.68 (d, 1 H), 7.41 (m, 6H), 5.17 (dd, 1 H), 4.42 (m, 2H), 4.24 (s, 2H), 3.63 (m, 4H), 3.38 (m, 4H), 2.98 (s, 3H), 2.97 (m, 2H). ES-MS: 452 [M+H]+.

[00173] 2-{3-[4-(4-Fluorophenyl)-1 ,2,3,6-tetrahydropyridin-1 -yl]-1 - methoxypropyl}-8-methoxy-3,4-dihydroquinazolin-4-one (125)

[00174] Prepared using General Procedure A, from compound 10 and 4-(4- Fluorophenyl)-1 ,2,3,6-tetrahydropyridine. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.80 (d, 1 H), 7.41 (dd, 1 H), 7.32 (d, 2H), 7.22 (d, 1 H), 6.98 (d, 2H), 5.91 (d, 1 H), 4.48 (dd, 1 H), 4.03 (s, 3H), 3.42 (s, 3H), 3.24 (m, 2H), 2.74 (m, 2H), 2.49 (m, 3H), 2.19 (m, 3H). ES-MS: 424 [M+H]+.

[00175] 2-{3-[4-(4-Fluorophenyl)-1 ,2,3,6-tetrahydropyridin-1 -yl]-1 -methoxy-

3-oxopropyl}-8-methoxy-3,4-dihydroquinazolin-4-one (126)

[00176] Prepared using General Procedure A, from compound 10 and 4-(4- Fluorophenyl)-1 ,2,3,6-tetrahydropyridine. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 10.02 (s, 1 H), 7.81 (d, 1 H), 7.41 (dd, 1 H), 7.32 (d, 2H), 7.22 (d, 1 H), 6.98 (d, 2H), 5.92 (d, 1 H), 4.92 (dd, 1 H), 4.25 (d, 1 H), 4.13 (d, 1 H), 4.03 (s, 3H), 3.82 (m, 1 H), 3.63 (m, 1 H), 3.52 (s, 3H), 2.94 (m, 2H), 2.52 (m, 2H). ES-MS: 438 [M+H]+.

[00177] 2-{3-[4-(4-Fluorophenyl)-1 ,2,3,6-tetrahydropyridin-1 -yl]-1 -hydroxy-

3-oxopropyl}-8-hydroxy-3,4-dihydroquinazolin-4-one (127)

[00178] Prepared using General Procedure B from compound 126. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.64 (d, 1 H), 7.40 (m, 3H), 7.22 (d, 1 H), 7.09 (d, 2H), 6.14 (d, 1 H), 5.12 (dd, 1 H), 4.22 (m, 2H), 3.83 (m, 2H), 3.11 (m, 3H), 2.62 (m, 1 H). ES-MS: 410 [M+H]+.

[00179] 8-Methoxy-2-(1 -methoxy-3-oxo-3-(4-phenylpiperazin-1 - yl)propyl)quinazolin-4(3/7)-one (128)

[00180] Prepared using General Procedure A, from compound 10 and 1- Phenylpiperazine. Purified by column chromatography eluting with a gradient of 3-30% MeOH in DCM, followed by preparative TLC eluting with 10% MeOH in DCM. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 10.05 (br s, 1 H), 7.91 (d, 1 H), 7.45 (t, 1 H), 7.31 (m, 1 H), 7.24 (m, 1 H), 6.93 (m, 4H), 4.94 (br m, 1 H), 4.02 (s, 3H), 3.81 (br m, 2H), 3.62 (br m, 2H), 3.56 (s, 3H), 3.18 (br m, 4H), 3.09-2.84 (br m, 2H). ES-MS: 423 [M+H]+.

[00181 ] 8-Hydroxy-2-(1 -methoxy-3-oxo-3-(4-phenylpiperazin-1 - yl)propyl)quinazolin-4(3/7)-one (129)

[00182] Prepared by General Procedure C from compound 128. Purified by column chromatography. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.79 (d, 1 H), 7.52 (t, 1 H), 7.43 (d, 1 H), 7.29 (t, 2H), 7.06 (d, 2H), 6.95 (t, 1 H), 5.18 (br m, 1 H), 4.02 (s, 3H), 3.80 (br m, 4H), 3.20 (br m, 4H), 2.71 (s, 2H). ES-MS: 409 [M+H]+.

[00183] Preparation of 8-Hydroxy-2-(3-(4-(4-hydroxy-3-

(hydroxymethyl)phenyl)piperazin-1 -yl)-1 -methoxy-3-oxopropyl)quinazolin- 4(3H)-one (133)

[00185] A solution of tert-butyl 4-(2,2-dimethyl-4/7-benzo[d][1 ,3]dioxin-6- yl)piperazine-1 -carboxylate 130 in 5 mL of DCM/TFA (1 :1 ) was stirred at ambient temperature for 1 h. All volatiles were removed by rotational evaporation to afford a crude red oil, which was combined with pTsOH’F (0.1 equiv.), 2,2- dimethoxypropane (5 equiv.) and Na2SO4 (excess) in anhydrous acetone. The mixture was stirred at ambient temperature for 4 h and then heated to 40 °C for 18.5 h. The solution was cooled ambient temperature, adsorbed onto silica and purified by column chromatography, eluting with DCM/MeOH/NH4OH (85/15/2) to yield amine 131. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 6.90 (d, 1 H), 6.73 (m, 2H), 4.82 (s, 2H), 3.36 (m, 4H), 3.28 (m, 4H), 1.49 (s, 6H). ES-MS: 249 [M+H]+.

[00186] 2-(3-(4-(2,2-Dimethyl-4H-benzo[d][1 ,3]dioxin-6-yl)piperazin-1 -yl)-1 - methoxy-3-oxopropyl)-8-methoxyquinazolin-4(3/7)-one (132)

[00187] Prepared using General Procedure A, from compound 10 and compound 131. Purified using column chromatography eluting with a gradient of 3- 10% MeOH in DCM. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.94 (br s, 1 H), 7.86 (d, 1 H), 7.42 (t, 1 H), 7.21 (d, 1 H), 6.76 (m, 2H), 6.52 (s, 1 H), 4.94 (t, 1 H), 4.79 (s, 2H), 3.98 (s, 3H), 3.77 (m, 2H), 3.63 (m, 2H), 3.51 (s, 3H), 2.99 (m, 6H), 1.51 (s, 6H). ESMS: 509 [M+H]+. [00188] 8-Hydroxy-2-(3-(4-(4-hydroxy-3-(hydroxymethyl)phenyl)piperazin-

1 -y I )-1 -methoxy-3-oxopropyl)quinazolin-4(3/7)-one (133)

[00189] Prepared using General Procedure C from compound 132. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.69 (d, 1 H), 7.49 (d, 1 H), 7.40 (t, 1 H), 7.30 (m, 2H), 6.92 (d, 1 H), 5.17 (t, 1 H), 4.51 (s, 2H), 4.02 (m, 4H), 3.58 (m, 4H), 3.46 (s, 3H), 3.20 (m, 2H). ES-MS: 455 [M+H]+.

[00190] 8-Methoxy-2-[1 -methoxy-3-oxo-3-(2-oxo-3-azepanylamino)propyl]- 3/7-quinazolin-4-one (134)

[00191] Prepared using General Procedure A, from compound 10 and 3-Amino- 2-azepanone.¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 10.90 (s, 1 H), 7.91-7.81 (m, 1 H), 7.48-7.38 (m, 1 H), 7.25-7.19 (m, 1 H), 5.87-5.76 (m, 1 H), 4.68-4.57 (m, 1 H), 4.02 (s, 3H), 3.48 (s, 3H), 3.36-3.23 (m, 2H), 2.90-2.79 (m, 2H), 2.13-1.95 (m, 2H), 1 .93-1 .76 (m, 2H), 1 .64-1 .31 (m, 3H). LR ESI MS: m/z calcd for C19H24N4O5 [M+H]⁺, 389.41.

[00192] 8-Hydroxy-2-[1 -hydroxy-3-oxo-3-(2-oxo-3-azepanylamino)propyl]- 3/7-quinazolin-4-one (135)

[00193] Prepared using General Procedure B from compound 134. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.80-7.72 (m, 1 H), 7.63-7.53 (m, 1 H), 7.46-7.38 (m, 1 H), 5.40-5.29 (m, 1 H), 4.65-4.49 (m, 1 H), 3.27-3.21 (m, 2H), 1.64-1.54 (m, 2H), 1.48- 1.12 (m, 4H). LR ESI MS: m/z calcd for C17H20N4O5 [M+H]⁺, 361.14.

[00194] Preparation of 2-(3-{2,6-Diazaspiro[3.5]nonan-6-yl}-1 - hydroxypropyl)-8-hydroxy-3,4-dihydroquinazolin-4-one (138)

[00195] Prepared using General Procedure A, from 10 and te/Y-Butyl 2,6- diazaspiro[3.5]nonane-2-carboxylate (136), to give Boc-protected dimethoxy intermediate 137 which was submitted to General Procedure B to yield title compound 138. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.68 (d, 1 H), 7.42 (dd, 1 H), 7.28 (d, 1 H), 4.78 (dd, 1 H), 4.16 (m, 3H), 4.08 (m, 4H), 3.44 (m, 3H), 2.42 (m, 2H), 1.94 (m, 4H). ES-MS: 345 [M+H]+.

[00196] Preparation of 2-(3-{1 ,8-Diazaspiro[4.5]decan-1 -yl}-1 - methoxypropyl)-8-methoxy-3,4-dihydroquinazolin-4-one (141), and 2-(3-{1,8- Diazaspiro[4.5]decan-1 -yl}-1-hydroxypropyl)-8-hydroxy-3,4-dihydroquinazolin- 4-one (142)

[00197] tert-Butyl 1 -[3-methoxy-3-(8-methoxy-4-oxo-3H-quinazolin-2- yl)propyl]-1 ,8-diaza-8-spiro[4.5]decanecarboxylate intermediate (140)

[00198] Prepared using General Procedure A, from compound 10 and tert-Butyl 1 ,8-diazaspiro[4.5]decane-8-carboxylate. Used without further purification.

[00199] General Procedure D

[00200] To a solution of /V-Boc protected compound (1 equiv.) in a suitable solvent (such as DCM, 1 ,4-dioxane, EtOH, DMF) at 0 °C is slowly added 4N HCI solution in 1 ,4-dioxane (5-10 equiv.). The mixture is stirred at room temperature until all starting material is consumed (usually 1 -8h), then the solvents are removed under vacuo to yield the amine product as a HCI salt. Optionally, the free amine can be obtained from basic wash I organic extraction.

[00201 ] 2-( 3-{1 ,8-Diazaspiro[4.5]decan-1 -yl}-1 -meth oxy propyl )-8-methoxy-

3,4-dihydroquinazolin-4-one (141 )

[00202] Prepared using General Procedure D from compound 140. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.85 (d, 1 H), 7.62 (dd, 1 H), 7.52 (d, 1 H), 4.61 (dd, 1 H), 4.18 (m, 2H), 4.02 (s, 3H), 3.58 (m, 2H), 3.50 (s, 3H), 3.24 (m, 2H), 2.47 (m, 3H), 2.26 (m, 3H), 2.17 (m, 2H), 1.37 (m, 4H), 0.97 (m, 2H). ES-MS: 387 [M+H]+.

[00203] 2-(3-{1 ,8-Diazaspiro[4.5]decan-1 -yl}-1 -hydroxypropyl)-8-hydroxy-

3,4-dihydroquinazolin-4-one (142)

[00204] Prepared using General Procedure B from compound 140. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.68 (d, 1 H), 7.38 (dd, 1 H), 7.25 (d, 1 H), 4.78 (dd, 1 H), 3.50 (m, 6H), 3.16 (m, 2H), 2.33 (m, 8H), 2.08 (m, 2H). ES-MS: 359 [M+H]+.

[00205] C. Benzyloxy-2-aminobenzamide intermediate synthesis

Scheme 3: Synthesis of Aminobenzamide 20

[00206] General Procedure E

[00207] Hydroxybenzoic acid starting material 17 (1 equiv.) was dissolved in DMF, then benzyl chloride (2.4 equiv.) and K2CO3 (3 equiv.) were added, and the mixture was heated at 50 °C for 18h. After cooling down to room temperature, EtOAc was added, and the mixture was filtered and concentrated to a dark brown residue. The residue was redissolved in DCM, washed with saturated aqueous NaHCOs, dried over Na2SO4, filtered and concentrated to a brown solid or paste. This residue was further purified by DCM/MeOH trituration and precipitation at room temperature. After filtration and wash with MeOH, the off-white solid was dried to yield benzyl ester 18 (60-80% yield).

[00208] General Procedure F

[00209] Benzyl ester 18 (1 equiv.) was dissolved in 1 ,4-dioxane I MeOH mixture (1/1 , v/v). NaOH solid (1.5 equiv.) was added, and the mixture was heated at 70 °C until TLC showed complete reaction (16-24h). Volatiles were removed under vacuum and the resulting solid redissolved in water (at about 1 M). At 0 °C the solution was acidified to pH = 3 with 2N HCI. The resulting precipitate was filtered, washed with cold water and dried under vacuum to yield benzoic acid 19 as an off-white solid (>90% yield).

[00210] General Procedure G

[00211] Benzoic acid 19 (1 equiv.) was dissolved in DMF and 1 ,1 '- carbonyldiimidazole (CDI, 1 .2 equiv.) was added. The mixture was heated at 70 °C for 2h. After cooling down to 0 °C, concentrated (28-30%) aqueous NH4OH (30 equiv.) was added slowly, then stirring maintained at 0 °C for 30 min, then room temperature for 1 h, and eventually the mixture was heated at 50 °C for 18h. The mixture was concentrated to remove DMF and yield a yellow solution. The residue was diluted with water and pH was adjusted to 10 with aqueous 2N NaOH. The product was extracted from the aqueous layer with EtOAc multiple times. The pooled organic layers were subsequently washed multiple times with saturated aqueous Na2CO3, a pH~6 mixture of aqueous NaHCO3/NH4CI and brine. The organic layer was then dried over Na2SO4, filtered and concentrated to yield benzamide 20 as a beige to light-yellow solid (50- 80% yield).

[00212] Benzyl 2-amino-3-(benzyloxy)benzoate (143)

[00213] Prepared using General Procedure E, from 2-Amino-3-hydroxybenzoic acid. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.56 (d, 1 H), 7.48-7.30 (m, 10H), 6.92 (d, 1 H), 6.55 (t, 1 H), 6.07 (br s, 2H), 5.33 (s, 2H), 5.09 (s, 2H).

[00214] 2-Amino-3-(benzyloxy)benzoic acid (144)

[00215] Prepared using General Procedure F, from compound 143. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.57 (d, 1 H), 7.49-7.32 (m, 5H), 6.96 (d, 1 H), 6.58 (t, 1 H), 6.4-5.7 (br s, 1 H), 5.11 (s, 2H). ES-MS: 244 [M+H]+.

[00216] 2-Amino-3-(benzyloxy)benzamide (145)

[00217] Prepared using General Procedure G, from compound 144. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.73 (br s, 1 H), 7.52-7.45 (m, 2H), 7.44-7.29 (m, 3H), 7.21 (d, 1 H), 7.10 (br s 1 H), 6.97 (d, 1 H), 6.46 (t, 1 H), 6.28 (br s, 2H), 5.13 (s, 2H). ES-MS: 243.9 [M+H]+.

[00218] D. Preparation of PARP Inhibitors Library following Synthetic Route II

Scheme 4: Synthetic Route II

[00219] General Procedure H

[00220] To a solution of the benzamide 21 (1 equiv.) in dioxane was added dropwise the acyl chloride (1.5 equiv.). The reaction was stirred for 1 h then diluted with water and the product 22 isolated by filtration.

[00221] General Procedure J

[00222] The amide 22 (1 equiv.) was suspended in a 1 :1 mixture of EtOH/5% aqueous NaOH and the reaction was heated to 90 °C and stirred for 20 minutes, then cooled to room temperature. The cyclized product 23 was precipitated by addition of AcOH and isolated by filtration.

[00223] General Procedure K

[00224] The olefin 23 (1 equiv.) and amine (2 equiv.) were dissolved in MeOH. AcOH (0.1 equiv.) was added, and the reaction was heated to 75 °C and stirred overnight. The reaction was diluted with water and the resulting solid 24 isolated by filtration to yield amine 23.

[00225] General Procedure L

[00226] The benzyloxy protected compound 24 was dissolved in MeOH and Pd/C (5%) was added. The reaction vessel was pressurized with H2 gas (50 psi) then evacuated (3x), and then pressurized with H2 gas and stirred overnight. The mixture was filtered through Celite and concentrated to give the hydroxy product 25.

[00227] 2-(3-Chloropropionylamino)-3-anisamide (22a)

[00228] Prepared using General Procedure H, from Amino-3- methoxybenzamide (6). The product was isolated as a white solid. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 9.30 (br s, 1 H), 7.38 (br s, 1 H), 7.32-7.23 (m, 2H), 7.13 (d, 1 H), 7.07 (d, 1 H), 3.79 (t, 2H), 3.77 (s, 3H), 2.74 (t, 2H). ESI-MS: 257 ([M+H]⁺).

[00229] 8-Methoxy-2-vinyl-3H-quinazolin-4-one (23a)

[00230] Prepared using General Procedure J, from compound 22a. The product was isolated as a white solid. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 11 .96 (br s, 1 H), 7.91 (d, 1 H), 7.45 (t, 1 H), 7.25 (d, 1 H), 6.80 (dd, 1 H), 6.64 (d, 1 H), 5.93 (d, 1 H),

4.05 (s, 3H).

[00231 ] 8-Methoxy-2-(2-morpholinoethyl)quinazolin-4(3/7)-one (146)

[00232] General Procedure K was followed using compound 23a and morpholine. The product was isolated as a white solid. ¹H NMR (300 MHz, Chloroformed): 5 (ppm) 12.96 (br, 1 H), 7.55-7.72 (d, 1 H), 7.24-7.47 (m, 2H), 3.88 (s, 3H), 3.49- 3.64 (m, 4H), 2.64-2.89 (m, 4H), 2.30-2.47 (br, 4H). ES-MS: 290.8 [M+H]+.

[00233] 8-Hydroxy-2-(2-morpholinoethyl)quinazolin-4(3H)-one (147)

[00234] General Procedure B was followed from compound 146. The desired product was isolated as a white solid. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 12.12 (br s, 1 H), 7.73 (d, 1 H), 7.34 (t, 1 H), 7.25 (d, 1 H), 3.92-3.82 (m, 4H), 2.95-2.81 (m, 4H), 2.72-2.61 (m, 4H). ES-MS: 276.7 [M+H]+.

[00235] 6-(4-(2-(8-Methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)ethyl)piperazin-1 -yl)nicotinonitrile (148)

[00236] General Procedure K was used from compound 23a and 6-(1- Piperazinyl)nicotinonitrile. The product was isolated as a white solid. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 12.10 (br s, 1 H), 8.45-8.41 (m, 1 H), 7.88-7.81 (m, 1 H), 7.69-7.61 (m, 1 H), 7.43-7.35 (m, 1 H), 7.23-7.16 (m, 1 H), 6.67-6.60 (m, 1 H), 4.01 (s, 3H), 3.88-3.81 (m, 4H), 3.04-2.97 (m, 2H), 2.92-2.85 (m, 2H), 2.78-2.71 (m, 4H). LR ESI MS: m/z calcd for C21H23N6O2 [M+H]⁺, 391 .4 found 391 .8.

[00237] Alternative to Synthetic Route II

Scheme 5: Synthetic Route I l-b

[00238] 2-(2-Chloroethyl)-8-methoxyquinazolin-4(3H)-one (27)

[00239] To a solution of Methyl 2-amino-3-methoxybenzoate (26) (1 equiv.) in 1 ,4-dioxane was added Acrylonitrile (2 equiv.). After cooling in ice bath, cone. HCI (2 equiv.) was added slowly. The reaction mixture was stirred in sealed tube at 80 °C overnight. After completion of reaction, all solvents and volatiles were evaporated. Water and DCM were added, then the pH of the mixture was adjusted to ~7-8 by careful addition of aqueous ammonia solution at 0 °C. The mixture was filtered, and the filtrate was stirred in methyl t-butyl ether to give solid product which was filtered and dried to yield title compound 27. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.68- 7.60 (m, 1 H), 7.46-7.30 (m, 2H), 4.08-3.98 (m, 2H), 3.15-3.05 (m, 2H). ESI MS: m/z calcd for C11H11CIN2O2 [M+H]⁺, 239.67.

[00240] General Procedure M

[00241] To a solution of compound 27 (1 equiv.) and required Amine (1 equiv.) in acetonitrile was added K2CO3 (2 equiv.) and the mixture was heated at 80 °C for 16-24h. After completion of the reaction, water was added and extracted with ethyl acetate. Organic layer was dried over Na2SO4, filtered and evaporated to get crude product. Pure Amine 28 was obtained by flash column chromatography.

[00242] Preparation of 4-((4-(2-(8-hydroxy-4-oxo-3,4-dihydroquinazolin-2- yl)ethyl)piperazin-1 -yl)methyl)benzonitrile (154)

[00243] tert-Butyl 4-(4-cyanobenzyl)piperazine-1 -carboxylate (151 )

[00244] To a solution of p-(Bromomethyl)benzonitrile (150) (1 equiv.) in acetonitrile is added DI PEA (1 .5 equiv.) followed by tert-Butyl 1 -piperazinecarboxylate (149) (1 .5 equiv.). The mixture was stirred at room temperature for 18h. Aqueous wash I EtOAc extraction yields the amine 151. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.64-7.57 (m, 2H), 7.49-7.40 (m, 2H), 3.57-3.51 (m, 2H), 3.47-3.37 (m, 4H), 2.43- 2.31 (m, 4H), 1.45 (m, 9H). ESI MS: m/z calcd for C17H23N3O2 [M+H]⁺, 302.38.

[00245] 4-(Piperazin-1-ylmethyl)benzonitrile (152) [00246] General Procedure D was followed using compound 151. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.53-7.45 (m, 2H), 7.39-7.32 (m, 2H), 3.43 (s, 2H), 2.85-2.72 (m, 4H), 2.41-2.19 (m, 4H). ESI MS: m/z calcd forCi2Hi₅N₃ [M+H]⁺, 202.26.

[00247] 4-((4-(2-(8-Methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)ethyl)piperazin-1 -yl)methyl)benzonitrile (153)

[00248] General Procedure M was followed using compound 152 and compound

27. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 12.44 (s, 1 H), 7.86-7.78 (m, 1 H), 7.63- 7.56 (m, 2H), 7.48-7.42 (m, 2H), 7.40-7.31 (m, 1 H), 7.20-7.12 (m, 1 H), 3.99 (s, 3H), 3.59 (m, 2H), 2.98-2.88 (m, 2H), 2.86-2.77 (m, 2H), 2.74-2.48 (m, 8H). LR ESI MS: m/z calcd for C23H25N5O2 [M+H]⁺, 404.47.

[00249] 4-((4-(2-(8-Hydroxy-4-oxo-3,4-dihydroquinazolin-2- yl)ethyl)piperazin-1 -yl)methyl)benzonitrile (154)

[00250] General Procedure B was followed using compound 153. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.31 (s, 1 H), 7.78-7.68 (m, 2H), 7.67-7.53 (m, 3H), 7.41- 7.29 (m, 1 H), 7.29-7.18 (m, 2H), 3.75 (s, 2H), 3.58-3.45 (m, 2H), 3.31-3.10 (m, 6H), 2.92-2.72 (m, 4H). LR ESI MS: m/z calcd for C22H₂3N₅O2[M+H]⁺, 390.45.

[00251] Preparation of 4-(4-(2-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)ethyl)piperazine-1 -carbonyl)benzonitrile (159)

[00252] General Procedure N

[00253] The carboxylic acid partner, such as compound 155, (1 equiv.) was suspended in DMF at room temperature under inert atmosphere. A coupling agent (such as HATU, DCC, EDC-HCI, or EDC-HCI with 1 equiv. additional HOBt) was added (1.5 equiv.). If required, DIPEA (2-4 equiv.) was slowly added. The clear solution was stirred at RT for 30 min., then the amine partner, such as compound 149, (1-2 equiv.) dissolved in a minimum amount of DMF was added into the mixture via syringe. The mixture was stirred for 16-24h (until reaction was deemed complete) and the solvent was removed under vacuum. Aqueous NaHCO₃ solution was added, and the mixture was stirred for 15 minutes after which, if precipitation occurred, the solid amide product was filtered off and dried. If no precipitation occurred, the compound was extracted with a suitable organic solvent (EtOAc, DCM for example), dried with Na2SO4, filtered and concentrated to give the amide product, such as compounds 156. Flash chromatography on silica gel was performed if further purification was required.

[00254] tert-Butyl 4-(4-cyanobenzoyl)piperazine-1 -carboxylate (156)

[00255] General Procedure N was followed using amine 149 and carboxylic acid 155. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.96-7.88 (m, 2H), 7.62-7.57 (m, 2H), 3.69-3.52 (m, 2H), 3.48-3.14 (m, 6H), 1 .40 (m, 9H). ESI MS: m/z calcd forCi7H2iN₃O₃ [M+H]⁺, 316.36.

[00256] 4-(Piperazine-1-carbonyl)benzonitrile (157)

[00257] General Procedure D was followed using compound 156. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.50-7.41 (m, 2H), 7.28-7.20 (m, 2H), 3.58-3.35 (m, 2H), 3.20-2.93 (m, 2H), 2.80-2.44 (m, 4H). ESI MS: m/z calcd for CI₂HI₃N₃0I [M+H]⁺, 316.36.

[00258] 4-(4-(2-(8-Methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)ethyl)piperazine-1 -carbonyl)benzonitrile (158)

[00259] General Procedure M was followed using compound 157 and compound 27. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 12.1 (s, 1 H), 7.85-7.78 (m, 1 H), 7.76- 7.69 (m, 2H), 7.56-7.48 (m, 2H), 7.43-7.34 (m, 1 H), 7.24-7.17 (m, 1 H), 4.01 (s, 3H), 3.98- 3.84 (m, 2H), 3.60-3.41 (m, 2H), 3.06-2.84 (m, 4H), 2.81-2.49 (m, 4H). LR ESI MS: m/z calcd for C2₃H₂₃N₅O₃[M+H]⁺ , 418.46. [00260] 4-(4-(2-(8-Hydroxy-4-oxo-3,4-dihydroquinazohn-2- yl)ethyl)piperazine-1 -carbonyl)benzonitrile (159)

[00261] General Procedure B was followed using compound 158. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.78-7.69 (m, 2H), 7.56-7.44 (m, 3H), 7.26-7.16 (m, 1 H), 7.14-7.05 (m, 1 H), 3.78-3.62 (m, 2H), 3.41-3.26 (m, 2H), 2.93-2.78 (m, 4H), 2.70- 2.41 (m, 4H). LR ESI MS: m/z calcd for C22H2iN₅O₃[M+H]⁺ , 404.43.

[00262] 3-(Benzyloxy)-2-(3-chloropropionylamino)benzamide (22b)

[00263] Prepared using General Procedure H, from 2-Amino-3- (benzyloxy)benzamide (compound 145). The product was isolated as a greyish solid. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 9.37 (br s, 1 H), 7.51-7.13 (m, 9H), 7.08 (d, 1 H), 5.13 (s, 2H), 3.79 (t, 2H), 2.75 (t, 2H).

[00264] 8-(Benzyloxy)-2-vinyl-3H-quinazolin-4-one (23b)

[00265] Prepared using General Procedure J, from compound 22b. The product was isolated as a white solid. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.32 (brs, 1 H), 7.71-7.63 (m, 1 H), 7.56-7.48 (m, 2H), 7.46-7.29 (m, 5H), 6.62-6.50 (m, 2H), 5.85-5.75 (m, 1 H), 5.34-5.24 (m, 2H). ESI-MS: 279 ([M+H]⁺).

[00266] 8-(Benzyloxy)-2-(2-morpholinoethyl)quinazolin-4(3H)-one (160)

[00267] General Procedure K was followed using compound 23b and Morpholine. The product was isolated as a white solid. ¹H NMR (300 MHz, Chloroformed): 5 (ppm) 12.24 (br s, 1 H), 7.86 (d, 1 H), 7.48 (d, 2H), 7.43-7.25 (m, 4H), 7.16 (d, 1 H), 5.36 (s, 2H), 3.90-3.80 (m, 4H), 3.00 (t, 2H), 2.85 (t, 2H), 2.71 -2.62 (m, 4H). ES-MS: 366 [M+H]+.

[00268] 8-(Benzyloxy)-2-(2-(diethylamino)ethyl)quinazolin-4(3H)-one (161 )

[00269] General Procedure K was followed using compound 23b and Diethylamine. The desired product was isolated as a white solid. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.75-7.65 (m, 1 H), 7.50-7.30 (m, 7H), 5.15 (s, 2H), 3.5-3.48 (m, 2H), 3.1-3.0 (m, 2H), 2.9-2.8 (m, 4H), 1.1 -0.98 (m, 6H). ES-MS: 350 [M+H]+.

[00270] 2-(2-(Diethylamino)ethyl)-8-hydroxyquinazolin-4(3H)-one (162)

[00271] General Procedure L was followed using compound 161. The desired compound was isolated as a sticky brown solid. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.72-7.64 (m, 1 H), 7.44-7.33 (m, 1 H), 7.32-7.22 (m, 1 H), 3.63-3.49 (m, 2H), 3.26-3.10 (m, 4H), 3.10-3.05 (m, 2H), 1.45-1.26 (m, 6H). ES-MS: 262 [M+H]+.

[00272] 8-(Benzyloxy)-2-(2-(piperidin-1 -yl)ethyl)quinazolin-4(3/7)-one (163)

[00273] General Procedure K was followed using compound 23b and Piperidine. The product was isolated as a white solid. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.68-7.61 (m, 1 H), 7.54-7.46 (m, 2H), 7.44-7.28 (m, 5H), 5.27 (s, 2H), 2.82-2.66 (m, 4H), 2.45-2.34 (m, 4H), 1.54-1.42 (m, 4H), 1.42-1.31 (m, 2H). ES-MS: 364 [M+H]+.

[00274] 8-Hydroxy-2-(2-(piperidin-1 -yl)ethyl)quinazolin-4(3/7)-one (164)

[00275] General Procedure L was followed using compound 163. The desired product was isolated as a white solid. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 12.70 (br s, 1 H), 7.72 (d, 1 H), 7.37-7.19 (m, 2H), 2.92-2.71 (m, 4H), 2.69-2.48 (m, 4H), 1.85- 1.66 (m, 4H), 1.64-1.47 (m, 2H). ES-MS: 274 [M+H]+.

[00276] 8-(Benzyloxy)-2-(2-(4-phenylpiperidin-1-yl)ethyl)quinazolin-4(3H)- one (165)

[00277] General Procedure K was followed using compound 23b and 4- Phenylpiperidine. The desired compound was isolated as a white solid. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.87 (d, 1 H), 7.51-7.46 (m, 2H), 7.42-7.20 (m, 9H), 7.15 (d, 1 H), 5.38 (s, 2H), 3.33-3.22 (m, 2H), 3.06-2.98 (m, 2H), 2.95-2.86 (m, 2H), 2.69- 2.57 (m, 1 H), 2.36-2.24 (m, 2H), 2.03-1.93 (m, 4H). ES-MS: 440 [M+H]+.

[00278] 8-Hydroxy-2-(2-(4-phenylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one

(166)

[00279] General Procedure L was followed using compound 165. The desired product was isolated as a white solid. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.74 (d, 1 H), 7.39-7.21 (m, 7H), 3.30-3.20 (m, 2H), 2.95-2.84 (m, 4H), 2.70-2.56 (m, 1 H), 2.36-2.22 (m, 2H), 2.04-1.93 (m, 4H). ES-MS: 350 [M+H]+.

[00280] E. Preparation of PARP Inhibitors Library following Synthetic Route

III

Scheme 6: Synthetic Route III

[00281] 8-Methoxy-2-(oxiran-2-yl)quinazolin-4(3H)-one (30)

[00282] The olefin 23a (1 equiv.) was dissolved in CHCh at 50 °C and m-CPBA (meta-chloroperoxybenzoic acid, 4 equiv.) was added in portions over 2 minutes. The reaction was stirred at 50 °C for 3 hours then cooled to room temperature and diluted with CH2CI2, washed with aqueous NaHCOa and concentrated. The crude residue was purified by column chromatography eluting with 40-100% EtOAc/hexanes to give the epoxide product 30 as a white solid. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 9.20 (br s, 1 H), 7.88 (d, 1 H), 7.46 (t, 1 H), 7.26 (d, 1 H), 4.17 (d, 1 H), 4.06 (s, 3H), 3.23 (t, 1 H), 2.97 (dd, 1 H). APC-MS: 219 [M+H]+.

[00283] General Procedure P

[00284] The epoxide 30 (1 equiv.) was dissolved in a mixture of methanol or isopropanol and 1 ,2-dichloroethane. The required amine (2 equiv.) was added, and the reaction was heated to 60 °C and stirred overnight. The reaction mixture was concentrated, and the residue was purified by column chromatography to yield hydroxy-amine 31.

[00285] 2-(1-Hydroxy-2-morpholinoethyl)-8-methoxyquinazolin-4(3H)-one

(167)

[00286] The title compound was prepared using General Procedure P, from epoxide 30 and Morpholine. The product was purified by column chromatography eluting with 0-10% MeOH/DCM. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.89 (d, 1 H), 7.43 (t, 1 H), 7.23 (d, 1 H), 4.80 (dd, 1 H), 4.03 (s, 3H), 3.85-3.72 (m, 4H), 3.04 (dd, 1 H), 2.78-2.66 (m, 3H), 2.62-2.52 (m, 2H). ES-MS: 306 [M+H]+.

[00287] 2-(2-(5,6-Dihydropyridin-1 (2H)-yl)-1 -hydroxyethyl)-8- methoxyquinazolin-4(3/7)-one (168)

[00288] The title compound was prepared using General Procedure P, from epoxide 30 and 1 ,2, 3-6 tetrahydropyridine. The product was purified by column chromatography eluting with 0-10% MeOH/DCM. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.89 (d, 1 H), 7.43 (t, 1 H), 7.23 (d, 1 H), 5.96-5.88 (m, 1 H), 5.74-5.68 (m, 1 H), 5.10 (dd, 1 H), 4.02 (s, 3H), 3.55-3.46 (m, 4H), 3.21-3.03 (m, 3H), 2.48-2.41 (m, 2H). ES-MS: 302 [M+H]+.

[00289] 2-(1 -Hydroxy-2-(4-phenylpiperazin-1 -yl)ethyl)-8- methoxyquinazolin-4(3/7)-one (169)

[00290] The title compound was prepared using General Procedure P, from epoxide 30 and 4-Phenylpiperazine. The product was purified by column chromatography eluting with 0-10% MeOH/DCM. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.90 (d, 1 H), 7.44 (t, 1 H), 7.34-7.21 (m, 3H), 6.99-6.87 (m, 3H), 4.83 (t, 1 H), 4.03 (s, 1 H), 3.44-3.40 (m, 4H), 3.15 (dd, 1 H), 2.96-2.86 (m, 2H), 2.83-2.71 (m, 3H). ES-MS: 381 [M+H]+.

[00291] F. Preparation of PARP Inhibitors Library following Synthetic Route IV

Scheme 7: Synthetic Route IV

[00292] General Procedure Q

[00293] The carboxylic acid 32 (1 equiv.) was suspended in anhydrous DCM under a nitrogen atmosphere. DMF (1 drop) was then added, followed by oxalyl chloride (0.95 equiv.), and the reaction mixture was stirred for 2h under nitrogen at room temperature. All volatiles were removed by rotational evaporation to afford crude acetyl chloride 33, used without further purification.

[00294] General Procedure R

[00295] To a flask charged with 2-amino-3-methoxybenzamide 6 (1 equiv.) dissolved in a minimum amount of 1 ,4-dioxane, was added a substituted acetyl chloride 33 (3 equiv.) and the white slurry was stirred at ambient temperature for 21 h. Ethanol was added to fully dissolve the material, then water was added dropwise to precipitate the product out of solution. The white solid was collected by filtration, washed with water, then toluene, and dried under vacuum. The product was taken up a 1 :3 mixture of ethanol and 1 M NaOH, heated up to 100 °C, and stirred until completion (usually 2-16 h). The reaction mixture was cooled to ambient temperature and the pH was adjusted to 4 with concentrated HCI. The resulting precipitate was collected by filtration, washed with water and dried thoroughly under vacuum to yield methoxy-quinazolinone 34.

[00296] 4-(8-Methoxy-4-oxo-3,4-dihydroquinazolin-2-yl)benzoic acid (170)

[00297] Prepared using General Procedure R, from compound 6 and Methyl 4- chlorocarbonylbenzoate. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.67 (s, 1 H), 8.30- 8.22 (m, 2H), 8.11-8.04 (m, 2H), 7.74-7.68 (m, 1 H), 7.51-7.43 (m, 1 H), 7.43-7.36 (m, 1 H), 3.95 (s, 3H). LR ESI MS: m/z calcd for C16H12N2O4 [M-H]; 295.27.

[00298] 8-Methoxy-2-(naphthalen-1 -ylmethyl)quinazolin-4(3/7)-one (171 )

[00299] Prepared using General Procedure Q and Rfrom 1 -Naphthylacetic acid, and compound 6, using dioxane as a solvent. Purified by washing with methanol. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 8.25 (d, 1 H), 7.94 (d, 1 H), 7.84 (d, 1 H), 7.28-7.66 (m, 7H), 4.42 (s, 2H), 3.81 (s, 3H). ES-MS: 317 [M+H]+.

[00300] 8-Methoxy-2-(4-methylstyryl)quinazolin-4(3H)-one (172)

[00301] Prepared using General Procedure Q and R from 4-Methylcinnamic acid, and compound 6. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 12.10 (s, 1 H), 7.92- 7.79 (m, 1 H), 7.79-7.71 (m, 1 H), 7.58-7.48 (m, 2H), 7.46-7.37 (m, 4H), 7.37-7.28 (m, 1 H), 7.27-7.17 (m, 2H), 7.02-6.88 (m, 1 H), 4.02 (s, 3H), 2.36 (s, 3H). LR ESI MS: m/z calcd for C18H18N2O2 [M+H]⁺ , 293.33.

[00302] 2-(4-(Ethoxymethyl)phenyl)-8-methoxyquinazolin-4(3H)-one (173)

[00303] Prepared using General Procedure Q and R from 4- (Bromomethyl)benzoic acid, and compound 6. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 12.14 (s, 1 H), 8.37-8.22 (m, 2H), 8.18-8.06 (m, 1 H), 7.99-7.86 (m, 1 H), 7.64- 7.37 (m, 4H), 7.32-7.18 (m, 1 H), 4.62 (s, 2H), 4.06 (s, 3H), 3.69-3.48 (m, 2H), 1.37- 1 .20 (m, 3H). LR ESI MS: m/z calcd for C18H18N2O3 [M+H]⁺ , 311 .34.

[00304] 8-Methoxy-2-(1-(6-methoxynaphthalen-2-yl)ethyl)quinazolin-4(3H)- one (174)

[00305] Prepared using General Procedure Q and R from 2-(6-methoxy-2- naphthyl)propanoic acid, and compound 6. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.81-7.71 (m, 3H), 7.51 (d, 1 H), 7.28 (t, 1 H), 7.21 (s, 1 H), 7.14-7.08 (m, 3H), 4.06-3.96 (m, 1 H), 3.89 (s, 3H), 3.72 (s, 3H), 1.60 (d, 3H). ES-MS: 361 [M+H]+.

[00306] 8-Methoxy-2-(methoxy(phenyl)methyl)quinazolin-4(3H)-one (175)

[00307] Prepared using General Procedure Q and R from 2-Methoxy-2- phenylacetic acid, and compound 6, using DCM as a solvent. Purified by column chromatography using 2-5% MeOH in DCM. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 10.44 (br s, 1 H), 7.82 (d, 1 H), 7.53 (d, 2H), 7.39-7.23 (m, 4H), 7.15 (d, 1 H), 5.43 (s, 1 H), 3.96 (s, 3H), 3.49 (s, 3H). ES-MS: 297 [M+H]+.

[00308] 8-Methoxy-2-(3-(piperidin-1 -yl)propyl)quinazolin-4(3/7)-one (176)

[00309] Prepared using General Procedure Q and R from 4-(Piperidin-1 - yl)butanoic acid, and compound 6. Purified by column chromatography eluting with a mixture of DCM/MeOH/NH4OH (90/10/1). ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 12.2 (br s, 1 H), 7.75 (d, 1 H), 7.32 (t, 1 H), 7.14 (d, 1 H), 3.99 (s, 3H), 3.09 (m, 6H), 2.45 (m, 2H), 2.02 (br m, 4H), 1.58 (m, 4H). ES-MS: 302 [M+H]+.

[00310] 8-Hydroxy-2-(3-(piperidin-1 -yl)propyl)quinazolin-4(3/7)-one (177)

[00311] Prepared by General Procedure B from compound 176. Purified by preparative TLC eluting with a mixture of DCM/MeOH/NH4OH. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.62 (d, 1 H), 7.32 (t, 1 H), 7.21 (d, 1 H), 2.80 (m, 2H), 2.65 (m, 4H), 2.10 (m, 2H), 1.69 (m, 4H), 1.54 (m, 2H), 1.30 (m, 2H). ES-MS: 288 [M+H]+.

[00312] 2-( 1 -Cyclopenten-1 -yl)-8-methoxy-3/7-quinazolin-4-one (178)

[00313] Prepared using General Procedure Q and R from 1-Cyclopentene-1- carboxylic acid, and compound 6. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.79 (d, 1 H), 7.44 (t, 1 H), 7.35 (d, 1 H), 6.87 (s, 1 H), 4.03 (s, 3H), 2.97-2.88 (m, 2H), 2.72-2.63 (m, 2H), 2.14-2.02 (m, 2H). ES-MS: 243 [M+H]+.

[00314] 2-(1 -Cyclopenten-1 -yl)-8-hydroxy-3H-quinazolin-4-one (179)

[00315] Prepared using General Procedure B from compound 178. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.66 (d, 1 H), 7.35 (t, 1 H), 7.24 (d, 1 H), 6.91 (s, 1 H), 2.98- 2.89 (m, 2H), 2.73-2.64 (m, 2H), 2.14-2.02 (m, 2H). ES-MS: 229 [M+H]+.

[00316] 8-Methoxy-2-(2-piperidinocyclopentyl)-3/7-quinazolin-4-one (180)

[00317] Prepared using General Procedure K from compound 178, using Piperidine as solvent and heating at 120 °C for 18h. ¹H NMR (300 MHz, DMSO-d6): 6 (ppm) 7.63 (d, 1 H), 7.41-7.28 (m, 3H), 3.90 (s, 3H), 3.46-3.37 (m, 1 H), 3.11-3.02 (m, 1 H), 2.53-2.43 (m, 2H), 2.42-2.31 (m, 2H), 2.06-1 .56 (m, 6H), 1.52-1.41 (m, 4H), 1.41- 1.31 (m, 2H). ES-MS: 328 [M+H]+.

[00318] Alternative to Synthetic Route IV

Scheme 8 : Synthetic Route IV-b

[00319] General procedure S

[00320] The open acid 35 (1 equiv.) was suspended in EtOH or MeOH at RT and 3M NaOH solution (~ 5 equiv.) was added slowly. After a few minutes, all solids dissolved, and a clear solution was observed. The clear solution was stirred for a further 2 to 5 hours when complete consumption of the open acid was confirmed. EtOH was removed and the resulting residue was dissolved in a minimum amount of water and carefully acidified to pH ~ 3-4 with slow addition of 2N aq. HCI. The resulting precipitated product was collected using suction filtration and dried to give cyclized quinazolinone 36 as a colorless solid (80-95% yield) which required no further purification. [00321] General procedure T

[00322] The benzylated species 37 was dissolved in a minimum amount of a suitable solvent (such as EtOH, MeOH, EtOAc, or DMF). A catalytic amount of Pd/C was added, and the reaction vessel was evacuated and backfilled with hydrogen twice. The mixture was stirred at room temperature, under a hydrogen atmosphere, for 5 to 12 hrs until complete consumption of the starting material. The mixture was filtered using celite and the celite cake was washed using MeOH. The combined filtrates were concentrated and optional purification using column chromatography gives the hydroxy-quinazolinone 38 as a colorless solid product (90-96% yield).

[00323] Preparation of p-[4-({[(8-Hydroxy-4-oxo-3H-quinazolin-2- yl)methyl]amino}carbonyl)-1 -piperidyl]benzonitrile (185)

[00324] General procedure D2

[00325] A solution of N-Boc protected compound in a 1 :2 (v:v) TFA/DCM mixture was stirred at room temperature until all starting material was consumed (usually 16- 24h). The mixture was concentrated to yield the deprotected Amine compound as a trifluoroacetate salt. Optionally, the free-base Amine compound could be obtained from basic treatment with NaOH and aqueous wash I organic extraction, followed by drying.

[00326] 2-[2-(Benzyloxy)-6-carbamoylphenylamino]-2-oxoethylamino 2,2- dimethylpropionate (181)

[00327] Synthesized using General Procedure N from compound 145 and N- Boc-Glycine. The product was purified by flash chromatography. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.29 (br s, 1 H), 7.42-7.28 (m, 5H), 7.23-7.17 (m, 2H), 7.08- 7.01 (m, 1 H), 6.31 (br s, 1 H), 5.70 (br s, 1 H), 5.23 (br s, 1 H), 5.12 (s, 2H), 3.94 (d, 2H), 1 .44 (s, 9H). LR ESI MS: m/z calcd for C21 H24N3O5 [M-Hp, 398.43 found 398.9.

[00328] 8-(Benzyloxy)-2-[(tert-butoxycarbonylamino)methyl]-3H- quinazolin-4-one (182)

[00329] Synthesized using General Procedure S from compound 181. Product was isolated as a white solid. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.81 (d, 1 H), 7.47-7.41 (m, 2H), 7.39-7.25 (m, 5H), 7.17 (d, 1 H), 5.30 (s, 2H), 4.28 (s, 2H), 1.45 (s, 9H). LR ESI MS: m/z calcd for C21 H22N3O4 [M-Hp, 380.42 found 380.8.

[00330] 2-(Aminomethyl)-8-(benzyloxy)-3H-quinazolin-4-one, trifluoroacetate salt (183)

[00331] Synthesized using General Procedure D2 from compound 182. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.83-7.78 (m, 1 H), 7.56-7.45 (m, 4H), 7.45-7.31 (m, 4H), 5.32 (s, 2H), 4.18 (s, 2H). LR ESI MS: m/z calcd for C16H16N3O2 [M+H]⁺, 282.32 found 282.5.

[00332] p-{4-[({[8-(Benzyloxy)-4-oxo-3H-quinazolin-2- yl]methyl}amino)carbonyl]-1 -piperidyl}benzonitrile (184)

[00333] Synthesized using General Procedure N from compound 183 and 1-(p- Cyanophenyl)-4-piperidinecarboxylic acid. The product was purified by flash chromatography. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.80 (d, 1 H), 7.47-7.14 (m, 10H), 6.80 (d, 2H), 5.26 (s, 2H), 4.32 (s, 2H), 3.85-3.75 (m, 2H), 2.91-2.79 (m, 2H), 2.49-2.33 (m, 1 H), 1.96-1.70 (m, 4H). LR ESI MS: m/z calcd for C29H28N5O3 [M+H]⁺, 494.56 found 494.8.

[00334] p-[4-({[(8-Hydroxy-4-oxo-3H-quinazolin-2- yl)methyl]amino}carbonyl)-1 -piperidyl]benzonitrile (185)

[00335] Synthesized using General Procedure T from compound 184. Semi- Prep HPLC purification gave the title compound. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.73-7.65 (m, 1 H), 7.59-7.48 (m, 2H), 7.33 (t, 1 H), 7.21 (d, 1 H), 7.02 (d, 2H), 4.28 (s, 2H), 4.14-3.90 (m, 3H), 2.99-2.87 (m, 2H), 1.86-1.75 (m, 2H), 1.71-1.57 (m, 2H). LR ESI MS: m/z calcd for C22H20N5O3 [M-Hp, 402.43, found 402.8.

[00336] Preparation of 6-{4-[2-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)acetyl]- 1 -piperazinyl}nicotinonitrile (190)

[00337] Methyl 3-[4-(5-cyano-2-pyridyl)-1 -piperazinyl]-3-oxopropionate (186)

[00338] To a solution of 6-(1-Piperazinyl)nicotinonitrile (1 equiv.) in DCM at 0 °C under inert atmosphere is added DI PEA (1 equiv.) and Methyl (chloroformyl)acetate (1 .2 equiv.). The mixture was stirred 2h at 0 °C then at room temperature for 16h. After addition of aq. NaHCOs, the product was extracted with DCM and concentrated to yield the title compound as a brown solid residue. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.43 (s, 1 H), 7.66 (d, 1 H), 6.62 (d, 1 H), 3.84-3.74 (m, 7H), 3.72-3.65 (m, 2H), 3.62-3.56 (m, 2H), 3.53 (s, 2H).

[00339] 3-[4-(5-Cyano-2-pyridyl)-1-piperazinyl]-3-oxopropionic acid (187)

[00340] To a solution of compound 186 (1 equiv.) in TH F/water mixture (4:1 , v:v) at 0 °C was added UOH-H2O (1 equiv.). The mixture was stirred 2h at 0 °C then at room temperature for 16h. Concentration yielded the title compound as a light yellow solid. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.44 (s, 1 H), 7.77 (d, 1 H), 6.89 (d, 1 H), 3.88-3.77 (m, 4H), 3.77-3.66 (m, 4H), 3.41 (s, 2H). LR ESI MS: m/z calcd for C13H15N4O3 [M+H]⁺, 275.28 found 275.8.

[00341] 3-(Benzyloxy)-2-{3-[4-(5-cyano-2-pyridyl)-1-piperazinyl]-3- oxopropionylamino}benzamide (188) [00342] Synthesized using General Procedure N from compound 187 and compound 145. Flash chromatography purification yielded the title compound as a light yellow solid. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 9.28 (br s, 1 H), 8.42 (s, 1 H), 7.64 (d, 1 H), 7.46-7.17 (m, 7H), 7.07 (d, 1 H), 6.55 (d, 1 H), 6.22 (br s, 1 H), 5.67 (br s, 1 H), 5.12 (s, 2H), 3.78-3.56 (m, 8H), 3.54 (s, 2H). LR ESI MS: m/z calcd for C27H27N6O4 [M+H]⁺, 499.54 found 499.9.

[00343] 6-(4-{2-[8-(Benzyloxy)-4-oxo-3H-quinazolin-2-yl]acetyl}-1- piperazinyl)nicotinonitrile (189)

[00344] Synthesized using General Procedure S from compound 188. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.38 (s, 1 H), 7.78 (d, 1 H), 7.60 (d, 1 H), 7.47-7.40 (m, 2H), 7.38-7.16 (m, 4H), 7.11 (d, 1 H), 6.45 (d, 1 H), 5.29 (s, 2H), 3.91 (s, 2H), 3.78- 3.60 (m, 8H). LR ESI MS: m/z calcd for C27H25N6O3 [M+H]⁺, 481 .52 found 481 .9.

[00345] 6-{4-[2-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)acetyl]-1 - piperazinyl}nicotinonitrile (190)

[00346] Synthesized using General Procedure T from compound 189. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 8.52 (s, 1 H), 7.88 (d, 1 H), 7.52 (d, 1 H), 7.30 (t, 1 H), 7.17 (d, 1 H), 6.95 (d, 1 H), 3.86 (s, 2H), 3.80-3.59 (m, 8H). LR ESI MS: m/z calcd for C20H19N6O3 [M+H]⁺, 391 .40 found 391 .8.

[00347] Preparation of p-[4-({[(8-Hydroxy-4-oxo-3H-quinazolin-2- yl)methyl]amino}carbonyl)-1 -piperazinyl]benzonitrile (195)

[00348] Methyl {[4-(p-cyanophenyl)-1 -piperazinyl]carbonylamino}acetate (191)

[00349] To a stirred solution of Methyl glycinate hydrochloride (1 equiv.) in anhydrous THF was added b/s(4-Nitrophenyl) carbonate (1.1 equiv.), and DIPEA (2.5 equiv.) at 0 °C temperature. After this, the reaction mixture was cooled to ambient temperature and was stirred at room temperature for 2 hours. The crude residue was diluted with EtOAc and washed with water, brine and dried over anhydrous Na2SO4 and concentrated to obtain crude material which was purified by flash column chromatography by using 0 to 30% of ethyl acetate in hexane to afford the yellow gummy (4-Nitrophenyl)-carbamate intermediate which was directly used for next step without any further purification. To a stirred solution of (4-Nitrophenyl)-carbamate intermediate (1 equiv.) in anhydrous THF was added 1 -(4-Cyanophenyl)piperazine (1.1 equiv.), and DI PEA (2 equiv.) at 0 °C temperature. After this, the reaction mixture was cooled to ambient temperature and was stirred at room temperature for 16 hours. The crude residue was diluted with EtOAc and washed with water, brine and dried over anhydrous Na2SO4 and concentrated to obtain crude material which was purified by flash column chromatography by using 0 to 5% of methanol in dichloromethane to afford the light-yellow solid ester 191. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.55-7.48 (m, 2H), 6.88-6.80 (m, 2H), 5.07-4.95 (m, 1 H), 4.07-4.02 (m, 2H), 3.77 (s, 3H), 3.65-3.57 (m, 4H), 3.42-3.35 (m, 4H). LR ESI MS: m/z calcd for C15H19N4O3 [M+H]⁺, 303.3 found 303.8.

[00350] {[4-(p-Cyanophenyl)-1-piperazinyl]carbonylamino}acetic acid (192)

[00351] To a stirred solution of ester intermediate 191 (1 equiv.) in MeOH:THF:water (4:2:1 ) was added lithium hydroxide (3 equiv.) and the mixture was stirred at room temperature for 3 hours. The solvent was evaporated, and crude residue was extracted with EtOAc and the aqueous phase was acidified with 1 M HCI solution to adjust the pH ~3-4. The residue was extracted with 5% MeOH in dichloromethane, washed with water, brine and dried over anhydrous Na2SO4 and concentrated to obtain crude material afford the off-white acid intermediate 192. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.44-12.29 (br s, 1 H), 7.65-7.54 (m, 2H), 7.10- 6.95 (m, 2H), 3.71-3.65 (m, 2H), 3.50-3.42 (m, 4H), 3.39-3.30 (m, 4H). LR ESI MS: m/z calcd for C14H16N4O3 [M+H]⁺, 289.3 found 289.7.

[00352] p-{4-[({[8-(Benzyloxy)-4-oxo-3H-quinazolin-2- yl]methyl}amino)carbonyl]-1 -piperazinyl}benzonitrile (194)

[00353] General Procedure S was used to prepare compound 194 from compound 193, which was prepared using General Procedure N from compounds 192 and 145. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.12-12.06 (br s, 1 H), 7.69-7.64 (m, 1 H), 7.63-7.56 (m, 2H), 7.51-7.45 (m, 2H), 7.43-7.24 (m, 5H), 7.04-6.98 (m, 2H), 5.24 (s, 2H), 4.22-4.17 (m, 2H), 3.51-3.43 (m, 4H), 3.37-3.30 (m, 4H). LR ESI MS: m/z calcd for C28H27N6O3 [M+H]⁺, 495.5 found 495.9.

[00354] p-[4-({[(8-Hydroxy-4-oxo-3H-quinazolin-2- yl)methyl]amino}carbonyl)-1 -piperazinyl]benzonitrile (195)

[00355] General Procedure T was used to prepare compound 195 from compound 194. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.63-7.56 (m, 2H), 7.52-7.45 (m, 1 H), 7.29-7.21 (m, 1 H), 7.17-7.01 (m, 4H), 4.27-4.20 (m, 2H), 3.69-3.23 (m, 10H). LR ESI MS: m/z calcd for C21H19N6O3 [M-H]; 403.4 found 403.8.

[00356] Preparation of p-{4-[2-(8-Hydroxy-4-oxo-3H-quinazolin-2- yl)ethylsulfonyl]-1 -piperazinyl}benzonitrile (198)

[00357] 3-[4-(p-Cyanophenyl)-1 -piperazinylsulfonyl]propionic acid (196)

[00358] To a stirred solution of Methyl 3-(chlorosulfonyl)propionate (1 equiv.) in DCM was added 1-(4-Cyanophenyl)piperazine (1.2 equiv.), and DIPEA (1.5 equiv.) at 0°C. The resulting solution was warmed to ambient temperature and stirred for 5 hours. The reaction mixture was quenched by water and extracted in DCM, washed with water, brine and dried over anhydrous Na2SO4 and concentrated to obtain crude sulfonamide intermediate. The crude material was dissolved in MeOH:DCM (3:1 ), followed by the addition of 2M aqueous NaOH (5% volume, excess). The resulting solution was stirred at room temperature for 4 hours and neutralized with 1 M aq. HCI solution to adjust the pH ~3 and extracted by using 5% MeOH in DCM, washed with water, brine and dried over anhydrous Na2SO4 and concentrated to obtain crude acid intermediate 196. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.54 (br s, 1 H), 7.65-7.59 (m, 2H), 7.10-7.04 (m, 2H), 3.47-3.41 (m, 4H), 3.36-3.26 (m, 6H), 2.68-2.62 (m, 2H). LR ESI MS: m/z calcd for C14H18N3O4S [M+H]⁺, 324.4 found 324.7.

[00359] p-(4-{2-[8-(Benzyloxy)-4-oxo-3H-quinazolin-2-yl]ethylsulfonyl}-1- piperazinyl)benzonitrile (197)

[00360] Prepared using General Procedure N and S from compounds 196 and 145. Compound 197 was obtained as white solid after filtration and was directly used without further purification. LR ESI MS: m/z calcd for C28H28N5O4S [M+H]⁺, 530.6 found 530.7.

[00361 ] p-{4-[2-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)ethylsulfonyl]-1 - piperazinyl}benzonitrile (198)

[00362] Prepared using General Procedure T from compound 197. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.23 (br s, 1 H), 9.38 (br s, 1 H), 7.65-7.55 (m, 2H), 7.53- 7.45 (m, 1 H), 7.33-7.24 (m, 1 H), 7.21-7.14 (m, 1 H), 7.10-7.00 (m, 2H), 3.85-3.77 (m, 2H), 3.46-3.29 (m, 8H), 3.13-3.05 (m, 2H). LR ESI MS: m/z calcd for C21H22N5O4S [M+H]⁺ , 440.5 found 440.9.

[00363] Preparation of p-{4-[(E)-3-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)-2- butenoyl]-1 -piperazinyl}benzonitrile (201 )

[00364] General procedure U

[00365] The selected Amine (1 equiv.) and required cyclic Anhydride (1.25 equiv.) were suspended in a 2:1 (v.v) mixture of DCM/AcOH or 1 ,4-dioxane/AcOH, and the resulting clear solution was stirred under argon at room temperature. Sometimes there is formation of a precipitate product after a few minutes but stirring is continued until all starting materials have reacted (1 to 12 hrs). Volatiles were removed and the solid product was collected using suction filtration and washed several times using Et2O/Hexanes mixture. No further purification was required (80- 95% yield).

[00366] (E)-4-[4-(p-Cyanophenyl)-1-piperazinyl]-2-methyl-4-oxo-2-butenoic acid (199) [00367] Prepared using general procedure U from p-(1-Piperazinyl)benzonitrile and 3-Methyl-2,5-furandione. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.53 (d, 2H), 6.85 (d, 2H), 5.75 (s, 1 H), 3.74 (m, 2H), 3.42 (m, 2H), 3.20 (m, 4H), 2.12 (s, 3H).

[00368] p-(4-{(E)-3-[8-(Benzyloxy)-4-oxo-3H-quinazolin-2-yl]-2-butenoyl}-1- piperazinyl)benzonitrile (200)

[00369] Prepared using General Procedure N and S from compounds 199 and 145. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.83 (d, 1 H), 7.52 (m, 4H), 7.36 (m, 4H), 7.19 (dd, 1 H), 6.79 (d, 2H), 5.78 (s, 1 H), 5.31 (s, 2H), 3.85 (m, 4H), 3.34 (m, 4H), 2.11 (s, 3H).

[00370] p-{4-[(E)-3-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)-2-butenoyl]-1- piperazinyl}benzonitrile (201)

[00371] Benzyl-protected compound 200 (1 equiv.) was dissolved in DCM, cooled to 0 °C, and BCh (1 equiv.) was added slowly. The reaction mixture was stirred at 0 °C for 5 minutes then additionally for 15 minutes at room temperature. Solvent was removed under reduced pressure and the product was purified by HPLC using acetonitrile formic acid mixture, yielding title compound 201 as a white solid. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.72 (d, 1 H), 7.52 (d, 2H), 7.36 (m, 2H), 6.86 (d, 2H), 5.76 (s, 1 H), 3.83 (m, 2H), 3.88 (m, 4H), 3.36 (m, 4H). ES-MS: 414.8 [M-H]-.

[00372] Preparation of p-{4-[3-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)propyl]- 1 -piperazinyl}benzonitrile (206)

[00373] Methyl 4-[4-(p-cyanophenyl)-1-piperazinyl]butyrate (202)

[00374] To a solution of Methyl 3-formylpropionate (1 equiv.) in DCE, was added p-(1-Piperazinyl)benzonitrile (1 equiv.) and stirred for 15 minutes then NaBH(OAc)3 (2 equiv.) was added and the mixture was stirred at room temperature for 18h. The reaction was quenched with saturated NaHCOs solution and extracted with DCM, dried over sodium sulfate and concentrated under vacuum to give compound 202 as a greenish liquid (64% yield). ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.64 (d, 2H), 6.64 (d, 2H), 3.68 (s, 3H), 3.32 (m, 4H), 2.55 (m, 4H), 2.47 (m, 4H), 1.77 (m, 2H).

[00375] 4-[4-(p-Cyanophenyl)-1-piperazinyl]butyric acid hydrochloride (203)

[00376] Compound 202 (1 equiv.) was dissolved in MeOH and 1 N NaOH (1.2 equiv.) was added. Mixture was stirred at room temperature for 5 hours; and the solvent was removed under vacuum. Then 1 N HCI was added to neutralize the sodium salt. All volatiles were removed under vacuum and 5:95 MeOH/DCM mixture was added to separate the product from NaCI salt. Solvent was removed under vacuum and the product was dried under vacuum, yielding the desired compound 203 hydrochloride salt form, as a white solid (84% yield). ¹H NMR (300 MHz, Methanol- d4) 5 (ppm) 7.64 (d, 2H), 7.16 (d, 2H), 3.70 (m, 4H), 2.45 (m, 4H), 3.30 (m, 2H), 2.50 (m, 2H), 2.10 (m, 2H).

[00377] p-[4-(4-{2-[(Aminooxy)methyl]-6-(benzyloxy)phenylamino}-4- oxobutyl)-1 -piperazinyl]benzonitrile (204)

[00378] Prepared using General Procedure N from compounds 203 and 145. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.34 (m, 7H), 6.86 (dd, 1 H), 6.78 (m, 3H), 6.40 (dd, 1 H), 5.95 (br s, 2H), 4.92 (s, 2H), 3.17 (m, 4H), 2.42 (m, 4H), 2.26 (m, 4H), 1.72 (m, 2H). ES-MS: 498.9 [M+H]+.

[00379] p-(4-{3-[8-(Benzyloxy)-4-oxo-3H-quinazolin-2-yl]propyl}-1- piperazinyl)benzonitrile (205)

[00380] Prepared using General Procedure S from compound 204. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.02 (m, 1 H), 7.61 (m, 2H), 7.42 (m, 5H), 7.11 (d, 4H), 5.35 (d, 2H), 3.42 (m, 4H), 2.74 (m, 2H), 2.57 (m, 2H), 2.53 (m, 2H), 2.11 (m, 2H), 0.88 (m, 2H). ES-MS: 478.8 [M-H]-. [00381 ] p-{4-[3-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)propyl]-1 - piperazinyl}benzonitrile (206)

[00382] Prepared using General Procedure T from compounds 205. ¹H NMR (300 MHz, Methanol-cM + Chloroform-d): 5 (ppm) 8.45 (br s, 2H), 7.92 (dd, 1 H), 7.45 (d, 2H), 7.23 (m, 1 H), 7.11 (d, 1 H), 6.89 (d, 2H), 3.32 (m, 4H), 2.74 (m, 2H), 2.65 (m, 2H), 2.52 (m, 2H), 2.01 (m, 2H), 0.8 (m, 2H). ES-MS: 489.0 [M-H]-.

[00383] Preparation of p-(4-{[3-(8-Hydroxy-4-oxo-3H-quinazolin-2- yl)cyclobutyl]carbonyl}-1 -piperazinyl)benzonitrile (211 )

[00384] Methyl 3-{[4-(p-cyanophenyl)-1- piperazinyl]carbonyl}cyclobutanecarboxylate (207)

[00385] Prepared using General Procedure N from p-(1-Piperazinyl)benzonitrile

(1 equiv.) and 3-Methoxycarbonylcyclobutanecarboxylic acid (1 equiv.) in DMF. Purification by column chromatography using Hexane/EtOAc mixture 10-20% gave compound 207 as colourless liquid (63% yield). ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.52 (d, 2H), 6.86 (d, 2H), 4.74 (m, 2H), 3.67 (s, 3H), 3.52 (m, 2H), 3.30 (m, 4H), 3.12 (m, 2H), 2.64 (m, 2H), 2.44 (m, 2H).

[00386] 3-{[4-(p-Cyanophenyl)-1- piperazinyl]carbonyl}cyclobutanecarboxylic acid (208) [00387] Ester 207 (1 equiv.) was dissolved in MeOH, and 1 N NaOH (1 .5 equiv.) was added. The reaction mixture was stirred at room temperature for 5 hours. The mixture was neutralized with 1 N HCI and MeOH was removed under vacuum and the product was extracted with EtOAc (3x50 mL), dried over sodium sulfate and concentrated under vacuum to yield acid 208 as a white solid (88% yield) which was used in the next step without purification. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.45 (d, 2H), 6.90 (d, 2H), 3.60 (m, 2H), 3.45 (m, 2H), 3.30 (m, 4H), 3.05 (m, 2H), 2.40 (m, 4H).

[00388] 3-(Benzyloxy)-2-[(3-{[4-(p-cyanophenyl)-1 - piperazinyl]carbonyl}cyclobutyl)carbonylamino]benzamide (209)

[00389] Prepared using General Procedure N from compounds 208 and 145. The crude was purified by column chromatography using (20-50 %) hexane/EtOAc mixture to give compound 209 as a white solid. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.49 (m, 1 H), 7.30 (d, 2H), 7.24 (m, 4H), 7.03 (m, 2H), 6.92 (m, 1 H), 6.71 (d, 2H), 4.92 (s, 2H), 3.50 (m, 2H), 3.35 (m, 2H), 3.20 (m, 5H), 2.51 (m, 2H), 2.40 (m, 2H), 2.02 (m, 1 H). ES-MS: 536.8 [M+H]+.

[00390] p-[4-({3-[8-(Benzyloxy)-4-oxo-3H-quinazolin-2- yl]cyclobutyl}carbonyl)-1 -piperazinyl]benzonitrile (210)

[00391] Prepared using General Procedure S from compound 209. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.81 (dd, 1 H), 7.50 (d, 3H), 7.39 (m, 6H), 6.83 (d, 2H), 5.35 (s, 2H), 3.75 (m, 4H), 3.34 (m, 4H), 3.09 (m, 2H), 2.87 (m, 2H), 2.66 (m, 2H). ESMS: 518.8 [M-H]-.

[00392] p-(4-{[3-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)cyclobutyl]carbonyl}- 1 -piperazinyl)benzonitrile (211 )

[00393] Prepared using General Procedure T from compound 210. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.58 (dd, 1 H), 7.46 (d, 2H), 7.26 (m, 1 H), 7.16 (m, 1 H), 6.90 (d, 2H), 3.71 (m, 2H), 3.50 (m, 4H), 3.24 (m,2H), 2.90 (m, 1 H), 2.71 (m, 5H). ESMS: 430.7 [M+H]+. [00394] Alternative to Synthetic Route IV

Scheme 9 : Synthetic Route IV-c

[00395] General procedure V

[00396] To a stirred solution of compound 145 (1 equiv.) in EtOH was added a selected aromatic or heteroaromatic aldehyde (1.2 equiv.), and iodine (1.2 equiv.) at ambient temperature. The resulting solution was heated to 85 °C and stirred for 6 hours. The reaction mixture was then cooled to ambient temperature and quenched by addition of 5% sodium thiosulfate solution and the solvent was removed by rotary evaporation. The crude residue was suspended in water and the solid was filtered. The solid cake was washed with 25% EtOAc in hexanes and dried under vacuum to obtain the mixture of quinazolinone intermediate 39.

[00397] Preparation of 6-{4-[m-(8-Hydroxy-4-oxo-3H-quinazolin-2- yl)phenyl]-1 -piperazinyl}nicotinonitrile (215)

[00398] General procedure W

[00399] To a stirred solution of Amine (intermediate 213, 1 equiv.) in anhydrous DMF was added halo-aryl or halo-heteroaryl (6-Chloronicotinate, 1.3 equiv.), K2CO3 (3 equiv.), and DMAP (0.2 equiv.) at ambient temperature. The resulting suspension was heated to 110 °C and stirred for 24 hours. After this, the reaction mixture was cooled to ambient temperature and the solvent was removed by rotary evaporation. The crude residue was diluted with EtOAc and washed with water, brine and dried over anhydrous Na2SO4 and concentrated to obtain crude material which was purified by flash column chromatography by using 0-10% MeOH in DCM to afford the Aminoaryl intermediate (214).

[00400] 8-(Benzyloxy)-2-[fn-(1-piperazinyl)phenyl]-3H-quinazolin-4-one

(213)

[00401] General Procedure V from compound 145 and 1 -Boc-4-(3-

Formylphenyl)piperazine gave a mixture of Boc-protected intermediate 212 and Boc- deprotected intermediate 213. The mixture was dissolved in DCM and trifluoroacetic acid was added (3: 1 v:v) and stirred at room temperature for 6 h. The solvent was removed by rotary evaporation and the residue was washed with EtOAc to obtain the intermediate 213 as TFA salt. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.56 (br s, 1 H), 8.85-8.72 (m, 1 H), 7.83-7.19 (m, 11 H), 5.34 (s, 2H), 3.51-3.42 (m, 4H), 3.34- 3.24 (m, 4H). LR ESI MS: m/z calcd for C25H23N4O2 [M+H]+, 411 .5 found 411 .9.

[00402] 6-(4-{m-[8-(Benzyloxy)-4-oxo-3H-quinazolin-2-yl]phenyl}-1- piperazinyl)nicotinonitrile (214)

[00403] Prepared using General Procedure W from compound 213. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.56 (br s, 1 H), 8.63-8.43 (m, 1 H), 8.02-6.88 (m, 14H), 5.33 (s, 2H), 3.96-3.75 (m, 4H), 3.48-3.28 (m, 4H). LR ESI MS: m/z calcd for C31H25N6O2 [M+H]+ , 513.6 found 513.9.

[00404] 6-{4-[m-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)phenyl]-1 - piperazinyl}nicotinonitrile (215)

[00405] Prepared using General Procedure T from compound 214.¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.41 (br s, 1 H), 8.57-8.49 (m, 1 H), 8.01-7.84 (m, 3H), 7.60-7.50 (m, 1 H), 7.43-7.24 (m, 2H), 7.23-7.12 (m, 2H), 7.08-7.00 (m, 1 H), 3.94- 3.78 (m, 4H), 3.49-3.30 (m, 4H). LR ESI MS: m/z calcd for C24H19N6O2 [M+H]+ , 423.4 found 423.9. [00406] Preparation of p-{4-[6-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)-2- py ridy l]-1 -piperazinyl}benzonitrile (218)

[00407] 8-(Benzyloxy)-2-(6-bromo-2-pyridyl)-3H-quinazolin-4-one (216)

[00408] Prepared using General Procedure V from compound 145 and 6-Bromo- 2-pyridinecarbaldehyde. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 10.75 (br s, 1 H), 8.59-8.54 (m, 1 H), 7.99-7.93 (m, 1 H), 7.80-7.72 (m, 1 H), 7.67-7.63 (m, 1 H), 7.58- 7.52 (m, 2H), 7.47-7.28 (m, 5H), 5.36 (s, 2H). LR ESI MS: m/z calcd for C₂oHi3BrN₃02 [M+H]+ , 407.3 found 408.8.

[00409] p-(4-{6-[8-(Benzyloxy)-4-oxo-3H-quinazolin-2-yl]-2-pyridyl}-1- piperazinyl)benzonitrile (217)

[00410] Prepared using General Procedure W from compound 216. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 11.82 (br s, 1 H), 7.85-7.73 (m, 3H), 7.66-7.54 (m, 4H), 7.52-7.33 (m, 5H), 7.14-7.06 (m, 3H), 5.36 (s, 2H), 3.91-3.83 (m, 4H), 3.58-3.50 (m, 4H). LR ESI MS: m/z calcd for C₃I H27N₆O₂ [M+H]⁺ , 515.6 found 515.9.

[00411 ] p-{4-[6-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)-2-pyridyl]-1 - piperazinyl}benzonitrile (218)

[00412] Prepared using General Procedure T from compound 217.¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 11.66 (br s, 1 H), 9.70 (br s, 1 H), 8.20-8.13 (m, 1 H), 7.86- 7.76 (m, 1 H), 7.68-7.57 (m, 3H), 7.42-7.33 (m, 1 H), 7.29-7.21 (m, 1 H), 7.15-7.00 (m, 3H), 3.93-3.79 (m, 4H), 3.61-3.48 (m, 4H). LR ESI MS: m/z calcd for C24H19N6O2 [M+H]+ , 423.4 found 423.4.

[00413] G. Preparation of PARP Inhibitors Library following Synthetic Route V

[00414] 4-(2-Carbamoyl-6-methoxyphenylamino)-4-oxobutanoic acid (43a)

[00415] Prepared using General Procedure U from 2-Amino-3- methoxybenzamide (6) and Succinic anhydride. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.38-7.28 (m, 1 H), 7.23-7.14 (m, 2H), 3.88 (s, 3H), 2.76-2.63 (m, 4H). LR ESI MS: 265 [M-H]-.

[00416] 3-(8-Methoxy-4-oxo-3H-quinazolin-2-yl)propionic acid (44a)

[00417] Prepared using General Procedure S from compound 43a. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.67-7.58 (m, 1 H), 7.45-7.23 (m, 2H), 3.88 (s, 3H), 2.90- 2.69 (m, 4H). LR ESI MS: 247 [M-H]-.

[00418] 4-[2-(Benzyloxy)-6-carbamoylphenylamino]-4-oxobutyric acid (43b)

[00419] Prepared using General Procedure U from 2-Amino-3- (benzyloxy)benzamide (145) and Succinic anhydride. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.16 (s, 1 H), 9.28 (s, 1 H), 7.55-7.47 (m, 2H), 7.47-7.32 (m, 4H), 7.26 (d, 1 H), 7.23-7.17 (m, 1 H), 7.16-7.10 (m, 1 H), 5.16 (s, 2H), 2.60-2.46 (m, 4H). ES-MS: 341.9 [M-H]-.

[00420] 3-[8-(Benzyloxy)-4-oxo-3H-quinazolin-2-yl]propionic acid (44b)

[00421] Prepared using General Procedure S from compound 43b. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.25 (s, 2H), 7.66 (dd, 1 H), 7.56-7.49 (m, 2H), 7.44-7.29 (m, 5H), 5.28 (s, 2H), 2.91-2.83 (m, 2H), 2.83-2.75 (m, 2H). ES-MS: 325.8 [M+H]+ .

[00422] 2-{3-[2-(Cyclopropylamino)-2-oxoethylamino]-3-oxopropyl}-8- methoxy-3H-quinazolin-4-one (219)

[00423] Synthesized using General Procedure N from compound 44a and 2- Amino-ZV-cyclopropylacetamide. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.21 (s, 1 H), 8.18 (t, 1 H), 7.90-7.83 (m, 1 H), 7.63 (d, 1 H), 7.42-7.28 (m, 2H), 3.88 (s, 3H), 3.62 (d, 2H), 2.87-2.78 (m, 2H), 2.71-2.62 (m, 2H), 2.63-2.55 (m, 1 H), 0.63-0.53 (m, 2H), 0.39-0.31 (m, 2H). LR ESI MS: m/z calcd for C17H21 N4O4 [M+H]⁺, 345.37 found 345.8.

[00424] 2-{3-[2-(Cyclopropylamino)-2-oxoethylamino]-3-oxopropyl}-8- hydroxy-3H-quinazolin-4-one (220)

[00425] Synthesized using General Procedure B from compound 219. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.16 (s, 1 H), 9.16 (s, 1 H), 8.15 (t, 1 H), 7.88-7.82 (m, 1 H), 7.49 (d, 1 H), 7.27 (t, 1 H), 7.17 (d, 1 H), 3.61 (d, 2H), 2.90-2.82 (m, 2H), 2.77-2.70 (m, 2H), 2.62-2.53 (m, 1 H), 0.62-0.53 (m, 2H), 0.37-0.31 (m, 2H). LR ESI MS: m/z calcd for C16H19N4O4 [M+H]⁺, 331 .35 found 331 .7.

[00426] 8-Methoxy-2-(3-{[(5-methyl-3-isoxazolyl)methyl]amino}-3- oxopropyl)-3H-quinazolin-4-one (221 )

[00427] Synthesized using General Procedure N from compound 44a and (5- Methylisoxazol-3-yl)methylamine. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.21 (s, 1 H), 8.56 (t, 1 H), 7.63 (d, 1 H), 7.42-7.28 (m, 2H), 6.02 (s, 1 H), 4.24 (d, 2H), 3.87 (s, 3H), 2.90-2.80 (m, 2H), 2.70-2.61 (m, 2H), 2.29 (s, 3H). LR ESI MS: m/z calcd for C17H19N4O4 [M+H]⁺, 343.36 found 343.7.

[00428] 8-Hydroxy-2-(3-{[(5-methyl-3-isoxazolyl)methyl]amino}-3- oxopropyl)-3H-quinazolin-4-one (222)

[00429] Synthesized using General Procedure B from compound 221. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 8.52 (t, 1 H), 7.50 (d, 1 H), 7.28 (t, 1 H), 7.17 (d, 1 H), 5.90 (s, 1 H), 4.23 (d, 2H), 2.93-2.84 (m, 2H), 2.77-2.68 (m, 2H), 2.20 (s, 3H). LR ESI MS: m/z calcd for C16H17N4O4 [M+H]⁺, 329.33 found 329.7.

[00430] N-[(S)-1-Benzyl-2-hydroxyethyl]3-(8-methoxy-4-oxo-3,4-dihydro-2- quinazolinyl)propionamide (223)

[00431] Synthesized using General Procedure N from compound 44a and (S)-2- Amino-3-phenylpropanol. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.82-7.73 (m, 1 H), 7.57-7.42 (m, 1 H), 7.41-7.35 (m, 1 H), 7.18-7.03 (m, 5H), 4.22-4.10 (m, 1 H), 4.05 (s, 3H), 3.59-3.53 (m, 2H), 2.98-2.83 (m, 3H), 2.78-2.63 (m, 3H). LR ESI MS: m/z calcd. for C21 H22N3O4 [M-Hp 380.2, found 380.8.

[00432] Preparation of (S)-2-[3-(6,7-Dimethoxy-4-oxo-3,4-dihydro-2- quinazolinyl)propionylamino]glutaric acid (226)

[00433] 3-(6,7-Dimethoxy-4-oxo-3H-quinazolin-2-yl)propionic acid (224)

[00434] Synthesized using General Procedure U, from 2-Amino-4,5-dianisamide and Succinic anhydride, followed by General Procedure S. ¹H NMR (300 MHz, DMSO- d6) 5 (ppm) 13.3-11 .3 (br s, 1 H), 7.41 (s, 1 H), 7.08 (s, 1 H), 3.89 (s, 3H), 3.85 (s, 3H), 2.94-2.69 (m, 4H). LR ESI MS: m/z calcd for C13H15N2O8 [M+H]⁺ 279.1 , found 279.6.

[00435] Dibenzyl (S)-2-[3-(6,7-dimethoxy-4-oxo-3,4-dihydro-2- quinazolinyl)propionylamino]glutarate (225) [00436] Synthesized using General Procedure N from compound 224 and Dibenzyl (S)-2-aminoglutarate. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 10.99 (br s 1 H), 7.50 (s, 1 H), 7.24 (m, 10H), 7.02 (s, 1 H), 6.82-6.67 (m, 1 H), 5.09 (s, 2H), 5.00 (s, 2H), 4.72-4.56 (m, 1 H), 3.91 (s, 6H), 3.42 (br s, 2H), 2.91 (m, 2H), 2.80 (m, 2H), 2.43- 1 .91 (m, 3H). LR ESI MS: m/z calcd for C32H33N3O8 [M+H]⁺ 588.2, found 588.9.

[00437] (S)-2-[3-(6,7-Dimethoxy-4-oxo-3,4-dihydro-2- quinazolinyl)propionylamino]glutaric acid (226)

[00438] The di-ester compound 225 (1 equiv.) was suspended in MeOH at RT and 3M NaOH solution (2.5 equiv.) was added slowly. The solution was stirred for 8 hours when complete consumption of the di-ester was confirmed. MeOH was removed and the resulting residue was dissolved in a minimum amount of water and carefully acidified to pH 3-4 with slow addition of 2N aq. HCI. The resulting precipitated product was collected by filtration and dried to give di-acid 226 as a white solid which required no further purification. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.64 (s, 1 H), 7.18 (s, 1 H), 4.52-4.37 (m, 1 H), 4.05 (s, 3H), 4.00 (s, 3H), 3.27-3.16 (m, 2H), 3.16-2.95 (m, 2H), 2.49-2.34 (m, 2H), 2.30-2.08 (m, 1 H), 2.04-1.84 (m, 1 H). LR ESI MS: m/z calcd. for C18H21 N₃O₈ [M-H]- 406.1 , found 406.7.

[00439] A/-(2-((5-cyanopyridin-2-yl)(methyl)amino)ethyl)-3-(8-methoxy-4- oxo-1 ,2,3, 4-tetrahydroquinazolin-2-yl)-A/-methylpropanamide (227)

[00440] General Procedure W using 6-Bromonicotinonitrile and N1,N2- Dimethylethane-1 ,2-diamine gave the intermediate 6-(Methyl(2- (methylamino)ethyl)amino)nicotinonitrile. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.40-8.37 (m, 1 H), 7.61-7.54 (m, 1 H), 6.54-6.47 (m, 1 H), 3.79-3.71 (m, 2H), 3.12 (s, 3H), 2.88-2.80 (m, 2H), 2.47 (s, 3H). LR ESI MS: m/z calcd for C10H14N4 [M+H]⁺, 191.2. 6-(Methyl(2-(methylamino)ethyl)amino)nicotinonitrile intermediate was then used with compound 44a following General Procedure N to give title compound 227. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.38-8.17 (m, 1 H), 7.90-7.80 (m, 1 H), 7.46-7.33 (m, 1 H), 7.30-7.13 (m, 2H), 7.14-7.02 (m, 1 H), 6.46-6.28 (m, 1 H), 4.02 (s, 3H), 3.92-3.75 (m, 2H), 3.14-2.95 (m, 8H), 2.81-2.72 (m, 1 H), 2.66-2.50 (m, 2H). LR ESI MS: m/z calcd for C22H26N6O3 [M-H]¹ 421 .0.

[00441] Preparation of N-{4-[4-(N-Methylcarbamoyl)-2- nitrophenylamino]butyl}3-(8-methoxy-4-oxo-3,4-dihydro-2- quinazolinyl)propionamide (231)

[00442] 4-[4-(tert-Butoxycarbonylamino)butylamino]-3-nitrobenzoic acid (228)

[00443] Prepared using General Procedure W from 4-Fluoro-3-nitrobenzoic acid and /V-Boc-1 ,4-diaminobutane. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.41 (br s, 1 H), 8.64-8.57 (m, 1 H), 8.54-8.44 (m, 1 H), 7.99-7.88 (m, 1 H), 7.17-7.05 (m, 1 H), 6.89- 6.74 (m, 1 H), 3.48-3.35 (m, 2H), 3.01-2.87 (m, 2H), 1.67-1.41 (m, 4H), 1.36 (s, 9H). LR ESI MS: m/z calcd. for C16H23N3O6 [M -H]⁺ 352.74.

[00444] 4-[4-(N-Methylcarbamoyl)-2-nitrophenylamino]butylamino-tert- butylformylate (229)

[00445] Prepared using General Procedure N from compound 228 and Methylamine. ¹H NMR (300 MHz, DMSO-d6): 6 (ppm) 8.64-8.57 (br s, 1 H), 8.54-8.44 (br s, 1 H), 7.99-7.88 (m, 1 H), 7.17-7.05 (m, 1 H), 6.89-6.74 (m, 1 H), 3.46-3.22 (m, 4H), 3.02-2.84 (m, 3H), 1.67-1.41 (m, 4H), 1.36 (s, 9H). LR ESI MS: m/z calcd. for C17H26N4O5 [M-Boc+H]⁺ 285.8.

[00446] N-Methyl4-(4-aminobutylamino)-3-nitrobenzamide, trifluoroacetate salt (230) [00447] Prepared using General Procedure D2 from compound 229 ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 8.64 (br s, 1 H), 8.51-8.43 (m, 2H), 8.40-8.29 (m, 1 H), 8.07-7.92 (m, 1 H), 7.85-7.48 (m, 1 H), 7.21-7.05 (m, 1 H), 3.52-3.28 (m, 2H), 2.96-2.64 (m, 5H), 1.76-1.51 (m, 4H). LR ESI MS: m/z calcd. for C12H18N4O3 [M+H]⁺ 267.7.

[00448] A/-{4-[4-(A/-Methylcarbamoyl)-2-nitrophenylamino]butyl}3-(8- methoxy-4-oxo-3,4-dihydro-2-quinazolinyl)propionamide (231 )

[00449] Prepared using General Procedure N from compound 230 and compound 44a. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 8.61 (s, 1 H), 8.52-8.42 (m, 1 H), 8.36-8.27 (m, 1 H), 8.24-8.14 (m, 1 H), 8.02-7.90 (m, 1 H), 7.60-7.54 (m, 1 H), 7.32- 7.15 (m, 2H), 7.10-7.01 (m, 1 H), 3.85 (s, 3H), 3.41-3.20 (m, 4 H), 3.15-3.04 (m, 2H), 2.84-2.70 (m, 4H), 2.61-2.53 (m, 2H), 1.66-1.40 (m, 2H). LR ESI MS: m/z calcd. for C24H28N6O6 [M+H]+ 495.9.

[00450] tert-Butyl-2-{1 -[3-(8-methoxy-4-oxo-3,4-dihydro-2- quinazolinyl)propionyl]-4-piperidyl}ethylaminoformylate (232)

[00451] Prepared using General Procedure N from compound 44a and 4-(2-Boc- aminoethyl)piperidine.¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.66-7.52 (m, 1 H), 7.36-7.16 (m, 2H), 6.77 (br s, 1 H), 4.40-4.24 (m, 1 H), 3.98-3.78 (m, 4H), 3.05-2.67 (m, 8H), 1.78-1.18 (m, 15H), 1.17-0.78 (m, 2H). LR ESI MS: m/z calcd. for C24H34N4O5 [M+H]+ 457.9.

[00452] 2-{3-[4-(2-Aminoethyl)-1-piperidyl]-3-oxopropyl}-8-methoxy-3,4- dihydro-4-quinazolinone, hydrochloride salt (233)

[00453] Prepared using General Procedure D from compound 232. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 8.07 (br s, 1 H), 7.73-7.63 (m, 1 H), 7.60-7.45 (m, 2H), 4.39- 4.25 (m, 1 H), 3.97 (s, 3H), 3.92-3.77 (m, 1 H), 3.13-2.88 (m, 4H), 2.85-2.66 (m, 3H), 1.79-1.41 (m, 5H), 1.16-0.80 (m, 2H). LR ESI MS: m/z calcd. for C19H26N4O3 [M+H]⁺ 359.8.

[00454] A/-(2-{1-[3-(8-Methoxy-4-oxo-3,4-dihydro-2-quinazolinyl)propionyl]- 4-piperidyl}ethyl)p-cyanobenzamide (234)

[00455] Prepared using General Procedure N from compound 233 and 4- Cyanobenzoic acid. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 8.02-7.93 (m, 2H), 7.88-7.81 (m, 2H), 7.78-7.70 (m, 1 H), 7.49-7.31 (m, 2H), 4.56-4.43 (m, 1 H), 4.14-3.93 (m, 4H), 3.51-3.41 (m, 1 H), 3.19-2.84 (m, 6H), 2.73-2.56 (m, 1 H), 1.90-0.99 (m, 7H). LR ESI MS: m/z calcd. for C27H29N5O4 [M+H]+ 486.5.

[00456] A/-(2-{1-[3-(8-Methoxy-4-oxo-3,4-dihydro-2-quinazolinyl)propionyl]- 4-piperidyl}ethyl)3-(8-methoxy-4-oxo-3,4-dihydro-2-quinazolinyl)propionamide (235)

[00457] Prepared using General Procedure N from compound 233 and compound 44a.¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 12.17 (br s, 1 H), 7.96 (br s, 1 H), 7.68-7.55 (m, 2H), 7.42-7.21 (m, 4H), 4.34-4.19 (m, 1 H), 3.86 (s, 6H), 3.15-2.99 (m, 2H), 2.93-2.68 (m, 7H), 2.63-2.54 (m, 1 H), 2.41-2.24 (m, 1 H), 0.75-1.74 (m, 7H). LR ESI MS: m/z calcd. for CsiHseNeOe [M+H]+ 589.6.

[00458] Preparation of 2-(3-{4-[3-(8-Hydroxy-4-oxo-3H-quinazolin-2- yl)propionyl]-1-piperazinyl}-1-methoxypropyl)-8-methoxy-3/7-quinazolin-4-one (239)

[00459] tert-Butyl 4-{3-[8-(benzyloxy)-4-oxo-3H-quinazolin-2-yl]propionyl}- 1 -piperazinecarboxylate (236)

[00460] Prepared using General Procedure N from compound 44b, and tertBuhl 1 -piperazinecarboxylate. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 10.90 (s, 1 H), 7.84 (d, 1 H), 7.49-7.42 (m, 2H), 7.40-7.23 (m, 4H), 7.15 (d, 1 H), 5.35 (s, 2H), 3.70-3.60 (m, 2H), 3.50-3.40 (m, 6H), 3.21-3.13 (m, 2H), 2.93-2.84 (m, 2H), 1.47 (s, 9H). LR ESI MS: m/z calcd for C27H33N4O5 [M+H]⁺, 493.2 found 493.8.

[00461 ] 8-(Benzyloxy)-2-[3-oxo-3-(1 -piperazinyl)propyl]-3/7-quinazolin-4- one (237)

[00462] Prepared using General Procedure D from compound 236. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.83 (d, 1 H), 7.70-7.52 (m, 4H), 7.47-7.34 (m, 3H), 5.44 (s, 2H), 3.89-3.73 (m, 4H), 3.37-3.25 (m, 4H), 3.25-3.13 (m, 4H). LR ESI MS: m/z calcd for C22H25N4O3 [M+H]⁺, 393.2 found 393.8.

[00463] 8-(Benzyloxy)-2-(3-{4-[3-methoxy-3-(8-methoxy-4-oxo-3H- quinazolin-2-yl)propyl]-1-piperazinyl}-3-oxopropyl)-3/7-quinazolin-4-one (238)

[00464] Prepared using General Procedure A from compound 237 and compound 10. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.08 (br s, 1 H), 7.85 (m, 2H), 7.49-7.19 (m, 6H), 7.15 (d, 1 H), 5.32 (s, 2H), 4.45 (t, 1 H), 4.02 (s, 3H), 3.71-3.61 (m, 1 H), 3.58-3.48 (m, 1 H), 3.48-3.38 (m, 5H), 3.16-3.08 (m, 2H), 2.87-2.78 (m, 2H), 2.76-2.63 (m, 1 H), 2.55-2.40 (m, 5H), 2.15-2.04 (m, 2H). LR ESI MS: m/z calcd for C35H39N6O6 [M+H]⁺, 639.3 found 639.8.

[00465] 2-(3-{4-[3-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)propionyl]-1- piperazinyl}-1-methoxypropyl)-8-methoxy-3/7-quinazolin-4-one (239)

[00466] Prepared using General Procedure T from compound 238. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 10.88 (br s, 2H), 7.86 (d, 1 H), 7.70 (d, 1 H), 7.43 (t, 1 H), 7.36-7.18 (m, 3H), 4.50-4.43 (m, 1 H), 4.02 (s, 3H), 3.71-3.60 (m, 1 H), 3.56-3.32 (m, 6H), 3.10-3.00 (m, 2H), 2.85-2.63 (m, 3H), 2.53-2.36 (m, 5H), 2.16-2.02 (m, 2H). LR ESI MS: m/z calcd for C28H33N6O6 [M+H]⁺, 549.2 found 549.8.

[00467] 8-Methoxy-2-(3-{4-[(3-methyl-2-oxo-1 -pyridyl)methyl]-1 -piperidyl}-

3-oxopropyl)-3,4-dihydro-4-quinazolinone (240)

[00468] Prepared using General Procedure N from compound 44a, and 3- Methyl-1-[(piperidin-4-yl)methyl]-1 ,2-dihydropyridin-2-one. ¹H NMR (300 MHz, DMSO- d6) 6 (ppm) 12.16 (br s, 1 H), 7.69-7.54 (m, 1 H), 7.50-7.20 (m, 4H), 6.17-6.01 (m, 1 H), 4.40-4.23 (m, 1 H), 4.02-3.65 (m, 6H), 3.05-2.89 (m, 1 H), 2.21-1.90 (m, 4H), 1.61-1.41 (m, 2H), 1.27-0.94 (m, 2H). LR ESI MS: m/z calcd. for C24H28N4O4 [M+H]+, 435.8.

[00469] 8-Hydroxy-2-(3-{4-[(3-methyl-2 -oxo-1 -pyridyl )methyl]-1 -piperidyl}-

3-oxopropyl)-4(3/7)-quinazolinone (241 )

[00470] Prepared using General Procedure N, from compound 44b and 3- Methyl-1-[(piperidin-4-yl)methyl]-1 ,2-dihydropyridin-2-one, followed by General Procedure T. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.88-7.78 (m, 1 H), 7.32-7.27 (m, 1 H), 7.22-7.18 (m, 1 H), 7.17-7.10 (m, 1 H), 7.05-6.95 (m, 1 H), 6.17-6.01 (m, 1 H), 4.75-4.61 (m, 1 H), 3.95-3.77 (m, 2H), 3.69-3.58 (m, 1 H), 3.21-3.09 (m, 2H), 3.05-2.91 (m, 1 H), 2.88-2.77 (m, 2H), 2.65-2.48 (m, 1 H), 2.29-2.08 (m, 4H), 1.78-1.64 (m, 2H), 1 .29-1 .07 (m, 2H). LR ESI MS: m/z calcd. for C23H26N4O4 [M+H]+, 421 .8.

[00471] 2-[3-(8-Methoxy-4-oxo-3H-quinazolin-2-yl)propionyl]-1, 2,3,4- tetrahydroisoquinoline-6-carbonitrile (242)

[00472] Prepared using General Procedure N, from compound 44a and 1 ,2,3,4- Tetrahydroisoquinoline-6-carbonitrile. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.24- 12.12 (br s, 1 H), 7.74-7.57 (m, 3H), 7.46-7.23 (m, 3H), 4.84-4.59 (m, 2H), 3.85-3.61 (m, 5H), 3.07-2.76 (m, 6H). LR ESI MS: m/z calcd for C22H20N4O3 : 388.42; found [M+H]⁺, found 389.90.

[00473] 8-Methoxy-2-(3-oxo-3-(3-oxopiperazin-1-yl)propyl)quinazolin-

4(3H)-one (243)

[00474] Prepared using General Procedure N, from compound 44a and 2- Piperazinone. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.81-7.72 (m, 1 H), 7.47- 7.37 (m, 1 H), 7.33-7.24 (m, 1 H), 4.37 (s, 2H), 4.18 (s, 1 H), 4.04-3.95 (m, 3H), 3.87- 3.72 (m, 2H), 3.49-3.41 (m, 1 H), 3.14-2.88 (m, 4H). LR ESI MS: m/z calcd for C16H18N4O4 [M+H]⁺, 331.33.

[00475] 8-(Benzyloxy)-2-(3-oxo-3-(3-oxopiperazin-1-yl)propyl)quinazolin- 4(3H)-one (244)

[00476] Prepared using General Procedure N, from compound 44b and 2- Piperazinone. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.21 (s, 1 H), 7.97-8.18 (m, 1 H), 7.59-7.72 (m, 1 H), 7.22-7.55 (m, 7H), 5.25 (s, 2H), 4.09 (s, 1 H), 3.88 (s, 1 H), 3.48-3.66 (m, 2H), 3.06-3.22 (m, 2H), 2.78-2.95 (m, 4H). LR ESI MS: m/z calcd for C22H22N4O4 [M+H]⁺, 406.43.

[00477] 8-Hydroxy-2-(3-oxo-3-(3-oxopiperazin-1-yl)propyl)quinazolin- 4(3H)-one (245)

[00478] Prepared using General Procedure T from compound 244. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.00 (s, 1 H), 7.68-7.58 (m, 1 H), 7.38-7.27 (m, 1 H), 7.26- 7.17 (m, 1 H), 4.34 (s, 2H), 4.16 (s, 1 H), 3.92-3.82 (m, 1 H), 3.82-3.73 (m, 1 H), 3.48- 3.40 (m, 1 H), 3.16-2.88 (m, 5H). LR ESI MS: m/z calcd for C15H16N4O4 [M+H]⁺, 317.31.

[00479] A/-(1 -(3-(8-Methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)piperidin-4-yl)acetamide (246)

[00480] Prepared using General Procedure N, from compound 44a and /V- (Piperidin-4-yl)acetamide. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.18 (s, 1 H), 7.88-7.75 (m, 1 H), 7.44-7.24 (m, 2H), 4.23-4.06 (m, 1 H), 3.95-3.67 (m, 5H), 3.20- 3.06 (m, 2H), 2.90-2.67 (m, 6H), 1.87-1.36 (m, 5H), 1.42-1.05 (m, 4H). LR ESI MS: m/z calcd for C19H24N4O4 [M+H]⁺, 373.41 .

[00481 ] 2-( 1 -(3-(8-Methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)piperidin-4-yl)-N-methylacetamide (247)

[00482] Prepared using General Procedure N, from compound 44a and /V- Methyl-2-(piperidin-4-yl)acetamide. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.16 (s, 1 H), 7.78-7.68 (m, 1 H), 7.66-7.57 (m, 1 H), 7.42-7.24 (m, 2H), 4.38-4.21 (m, 1 H), 3.87 (s, 3H), 3.08-2.93 (m, 1 H), 2.87-2.74 (m, 5H), 2.00-1.78 (m, 3H), 1.74-1.48 (m, 2H), 1.21-1.01 (m, 1 H), 1.01-0.79 (m, 1 H). LR ESI MS: m/z calcd for C20H26N4O4 [M+H]⁺, 387.44.

[00483] tert-Butyl 4-(3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)-1 ,4-diazepane-1 -carboxylate (248)

[00484] Prepared using General Procedure N, from compound 44a and 1-Boc- 1 ,4-diazepane. ¹H NMR (300 MHz, Chloroform-d): 6 (ppm) 7.74-7.65 (m, 1 H), 7.31- 7.18 (m, 1 H), 7.12-7.01 (m, 1 H), 3.85 (s, 3H), 3.14-2.99 (m, 2H), 2.89-2.77 (m, 2H), 2.64-2.50 (m, 4H), 1.89-1.64 (m, 4H), 1.33 (s, 9H). LR ESI MS: m/z calcd for C22H30N4O5 [M+H]⁺, 431.49.

[00485] 2-(3-(1 ,4-Diazepan-1 -yl)-3-oxopropyl)-8-methoxyquinazolin-4(3/7)- one, hydrochloride salt (249)

[00486] Prepared using General Procedure D, from compound 248. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.71-7.58 (m, 2H), 7.58-7.38 (m, 2H), 7.31-7.18 (s, 1 H), 7.12-7.01 (m, 1 H), 3.94 (s, 3H), 3.78-3.62 (m, 2H), 3.60-3.43 (m, 2H), 3.36- 2.96 (m, 8H), 2.14-1.83 (m, 2H). LR ESI MS: m/z calcd for C17H23CIN4O3 [M+H]⁺, 331.38. [00487] 6-(4-(3-(8-Methoxy-4-oxo-3,4-dihydroquinazolin-2-yl)propanoyl)- 1 ,4-diazepan-1 -yl)nicotinonitrile (250)

[00488] Prepared using General Procedure W, from compound 249 and 6- Bromonicotinonitrile. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 11.22-10.78 (m, 1 H), 8.39-8.22 (m, 1 H), 7.86-7.77 (m, 1 H), 7.53-7.31 (m, 2H), 7.23-7.13 (m, 1 H), 6.56- 6.41 (m, 1 H), 3.99 (s, 3H), 3.86-3.71 (m, 2H), 3.71-3.63 (m, 1 H), 3.63-3.52 (m, 2H), 3.52-3.35 (m, 1 H), 3.22-2.98 (m, 2H), 2.96-2.81 (m, 1 H), 2.81-2.66 (m, 1 H), 2.10- 1.84 (m, 2H). LR ESI MS: m/z calcd for C23H24N6O3 [M+H]⁺, 433.47.

[00489] 8-Methoxy-2-(3-oxo-3-(2-oxo-1,7-diazaspiro[3.5]nonan-7- yl)propyl)quinazolin-4(3/7)-one (251 )

[00490] Prepared using General Procedure N, from compound 44a and 1 ,7- Diazaspiro[3.5]nonan-2-one. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.59-7.56 (m, 1 H), 7.37-7.27 (m, 1 H), 7.27-7.20 (m, 1 H), 3.89 (s, 3H), 3.66-3.35 (m, 4H), 2.90 (s, 4H), 2.64 (s, 2H), 1.87-1.55 (m, 4H). ESI MS: m/z calcd for C19H22N4O4 [M+H]⁺, 371.40.

[00491] 8-(Benzyloxy)-2-(3-oxo-3-(2-oxo-1,7-diazaspiro[3.5]nonan-7- yl)propyl)quinazolin-4(3H)-one (252)

[00492] Prepared using General Procedure N, from compound 44b and 1 ,7- Diazaspiro[3.5]nonan-2-one. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 8.31 (s, 1 H), 7.70-7.60 (m, 1 H), 7.54-7.45 (m, 2H), 7.44-7.27 (m, 5H), 5.25 (s, 2H), 3.60-3.37 (m, 4H), 2.97-2.78 (m, 4H), 2.62-2.55 (m, 2H), 1.78-1.43 (m, 4H). LR ESI MS: m/z calcd for C25H26N4O4 [M+H]⁺, 447.49.

[00493] 8-Hydroxy-2-(3-oxo-3-(2-oxo-1,7-diazaspiro[3.5]nonan-7- yl)propyl)quinazolin-4(3/7)-one (253)

[00494] Prepared using General Procedure T from compound 252. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.00 (s, 1 H), 7.68-7.57 (m, 1 H), 7.39-7.27 (m, 1 H), 7.27- 7.15 (m, 1 H), 3.83-3.48 (m, 4H), 3.15-2.95 (m, 4H), 2.75 (s, 2H), 2.01-1.68 (m, 4H). LR ESI MS: m/z calcd for C18H20N4O4 [M+H]⁺, 357.37.

[00495] 8-(Benzyloxy)-2-(3-oxo-3-(2-oxo-1,8-diazaspiro[4.5]decan-8- yl)propyl)quinazolin-4(3/7)-one (254)

[00496] Prepared using General Procedure N, from compound 44b and 2-Oxo- 1 ,8-diazaspiro[4.5]decane. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.78-7.18 (m, 3H), 5.23 (s, 2H), 3.64-3.40 (m, 4H), 3.08-2.70 (m, 6H), 2.11-1 .91 (m, 2H), 1 .60-1 .25 (m, 4H). ESI MS: m/z calcd for C26H28N4O4 [M+H]⁺, 461.52.

[00497] 8-Hydroxy-2-(3-oxo-3-(2-oxo-1,8-diazaspiro[4.5]decan-8- yl)propyl)quinazolin-4(3/7)-one (255)

[00498] Prepared using General Procedure T from compound 254. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.55-7.44 (m, 1 H), 7.26-7.13 (m, 1 H), 7.12-7.04 (m, 3H), 3.67-3.27 (m, 4H), 3.13-3.07 (m, 2H), 2.95-2.84 (m, 4H), 2.13-2.11 (m, 2H), 1.63- 1 .43 (m, 4H). ESI MS: m/z calcd for C19H22N4O4 [M+H]⁺, 371 .40. [00499] 8-Hydroxy-2-(3-{4-[(2-imidazolyl)methyl]-1-piperidyl}-3-oxopropyl)- 4(3/7)-quinazolinone (256)

[00500] Prepared using General Procedure N, from compound 44b and 4-((1/7- lmidazol-2-yl)methyl)piperidine, followed by General Procedure T. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.30-7.21 (m, 1 H), 7.14-7.04 (m, 1 H), 6.95-6.88 (m, 1 H), 6.85- 6.74 (m, 2H), 4.45-4.30 (m, 1 H), 4.10-3.83 (m, 1 H), 2.97-2.79 (m, 5H), 2.55-2.24 (m, 3H), 1.95-1.78 (m, 1 H), 1.61-1.42 (m, 1 H), 0.85-0.70 (m, 1 H). LR ESI MS: m/z calcd. for C20H23N5O3 [M+H]+ 380.6.

[00501] Preparation of 4-(4-(3-(8-Methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)piperazine-1 -carbonyl)benzonitrile (259)

[00502] tert-Butyl 4-(4-cyanobenzoyl)piperazine-1 -carboxylate (257)

[00503] To a solution of p-Cyanobenzoyl chloride (1 equiv.) in DCM at 0 °C is added DIPEA (1.5 equiv.) followed by tert-Butyl 1 -piperazinecarboxylate (149) (1.5 equiv.). The mixture was stirred at room temperature for 18h. Aqueous wash I EtOAc extraction yields the title compound. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.75- 7.68 (m, 2H), 7.53-7.46 (m, 2H), 3.82-3.21 (m, 8H), 1.45 (s, 9H). ESI MS: m/z calcd for C17H21N3O3 [M+H]⁺, 316.36.

[00504] 4-(Piperazine-1-carbonyl)benzonitrile, hydrochloride salt (258)

[00505] Prepared using General Procedure D, from compound 257. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.93-7.84 (m, 2H), 7.74-7.64 (m, 2H), 4.18-3.55 (m, 8H). ESI MS: m/z calcd for C12H14CIN3O [M+H]⁺, 216.25.

[00506] 4-(4-(3-(8-Methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)piperazine-1 -carbonyl)benzonitrile (259)

[00507] Prepared using General Procedure N, from compound 258 and compound 44a. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 10.81 (s, 1 H), 7.88-7.80 (m, 1 H), 7.78-7.69 (m, 2H), 7.55-7.48 (m, 2H), 7.43-7.33 (m, 1 H), 7.23-7.14 (m, 1 H), 4.00 (s, 3H), 3.21-3.12 (m, 8H), 2.98-2.79 (m, 2H). ESI MS: m/z calcd forC24H₂3N₅O4 [M+H]⁺, 446.47.

[00508] Preparation of 6-(1-(3-(8-Methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)piperidin-4-ylamino)nicotinonitrile (262)

[00509] tert-Butyl 4-(5-cyanopyridin-2-ylamino)piperidine-1 -carboxylate (260)

[00510] Prepared using General Procedure W, from 4-Amino-1-Boc-piperidine and 6-Chloronicotinonitrile. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.27 (s, 1 H), 7.51-7.41 (m, 1 H), 6.41-6.32 (m, 1 H), 5.46-5.35 (m, 1 H), 4.13-3.81 (m, 3H), 2.98-

2.78 (m, 2H), 2.03-1 .79 (m, 2H), 1 .41 (s, 9H), 1 .38-1 .25 (m, 2H). ESI MS: m/z calcd for C16H22N4O2 [M+H]⁺, 303.37.

[00511 ] 6-(Piperidin-4-ylamino)nicotinonitrile, hydrochloride salt (261 )

[00512] Prepared using General Procedure D, from compound 260. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.48 (s, 1 H), 8.05-7.91 (m, 1 H), 7.21-7.04 (m, 1 H), 4.25-4.04 (m, 1 H), 3.60-3.42 (m, 2H), 3.28-3.10 (m, 2H), 2.37-2.21 (m, 2H), 2.01-

1 .79 (m, 2H). ESI MS: m/z calcd for C11H15N4CI [M+H]⁺, 203.25.

[00513] 6-( 1 -(3-(8-Methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)piperidin-4-ylamino)nicotinonitrile (262)

[00514] Prepared using General Procedure N, from compound 261 and compound 44a. ¹H NMR (300 MHz, Chloroform-d): 6 (ppm) 8.31 (s, 2H), 7.87-7.71 (m, 1 H), 7.54-7.42 (m, 1 H), 7.40-7.29 (m, 1 H), 7.22-7.10 (m, 1 H), 6.48-6.31 (m, 1 H), 5.66-5.49 (m, 1 H), 4.61-4.44 (m, 1 H), 4.12-3.92 (m, 4H), 3.92-3.77 (m, 1 H), 3.30- 3.05 (m, 3H), 2.96-2.76 (m, 3H), 2.43-1.93 (m, 3H), 1.52-1.30 (m, 2H). ESI MS: m/z calcd for C23H24N₆O₃ [M+H]⁺, 433.47.

[00515] Preparation of 6-((1 -(3-(8-Hydroxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)piperidin-4-yl)(methyl)amino)nicotinonitrile (266)

[00516] tert-Butyl 4-((5-cyanopyridin-2-yl)(methyl)amino)piperidine-1 - carboxylate (263)

[00517] Prepared using General Procedure W, from 1-Boc-4- Methylaminopiperidine and 6-Chloronicotinonitrile. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.39-8.32 (m, 1 H), 7.62-7.53 (m, 1 H), 6.50-6.41 (m, 1 H), 4.89-4.69 (m, 1 H), 4.37-4.08 (m, 2H), 2.95-2.72 (m, 5H), 1.69-1.59 (m, 4H), 1.45 (s, 9H). ESI MS: m/z calcd for C17H24N4O2 [M+H]⁺, 317.39.

[00518] 6-(Methyl(piperidin-4-yl)amino)nicotinonitrile, hydrochloride salt (264)

[00519] Prepared using General Procedure D, from compound 263. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.61-8.56 (m, 1 H), 8.23-8.14 (m, 1 H), 7.71-7.56 (m, 1 H), 4.78-4.59 (m, 1 H), 3.66-3.53 (m, 2H), 3.42-3.27 (m, 2H), 3.25 (s, 3H), 2.39- 2.05 (m, 4H). ESI MS: m/z calcd for C12H17CIN4 [M+H]⁺, 217.28.

[00520] 6-((1 -(3-(8-(Benzyloxy)-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)piperidin-4-yl)(methyl)amino)nicotinonitrile (265)

[00521] Prepared using General Procedure N, from compound 264 and compound 44b. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.23 (s, 1 H), 8.52-8.44 (m, 1 H), 7.87-7.79 (m, 1 H), 7.70-7.62 (m, 1 H), 7.55-7.46 (m, 2H), 7.44-7.24 (m, 5H), 6.84-6.70 (m, 1 H), 5.26 (s, 2H), 3.13-2.96 (m, 3H), 2.95-2.83 (m, 5H), 2.80 (s, 3H), 1 .59-1 .46 (m, 5H). LR ESI MS: m/z calcd for C30H30N6O3 [M+H]⁺, 523.59.

[00522] 6-((1 -(3-(8-Hydroxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)piperidin-4-yl)(methyl)amino)nicotinonitrile (266)

[00523] Prepared using General Procedure T, from compound 265. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 13.10-10.80 (br s, 1 H), 10.21-8.73 (br s, 1 H), 8.44- 8.52 (m, 1 H), 7.89-7.78 (m, 1 H), 7.54-7.45 (m, 1 H), 7.32-7.22 (m, 1 H), 7.21-7.12 (m, 1 H), 6.85-6.71 (m, 1 H), 4.86-4.63 (m, 1 H), 4.60-4.40 (m, 1 H), 4.23-4.02 (m, 1 H), 3.06-2.93 (m, 2H), 2.92-2.82 (m, 5H), 1.91-1.37 (m, 4H). LR ESI MS: m/z calcd for C23H24N6O3 [M+H]⁺, 433.47.

[00524] Preparation of p-({4-[3-(8-Hydroxy-4-oxo-3H-quinazolin-2- yl)propionyl]-1 -piperazinyl}methyl)benzonitrile (268)

[00525] p-[(1-Piperazinyl)methyl]benzonitrile, hydrochloride salt (267)

[00526] Compound ferf-Butyl 1 -piperazinecarboxylate (1 equiv.) and K2CO3 (2 equiv.) were mixed in CH3CN and p-(Bromomethyl)benzonitrile (1 equiv.) was added slowly, the mixture was stirred at room temperature for 18h. The reaction was quenched with 5N HCI solution (excess) and stirred for 5 minutes, CH3CN was removed under vacuum and EtOAc was added, the organic layer was separated, and aqueous layer was washed with EtOAc, and the aqueous layer was then neutralized with Na2CO3 solution. The product was extracted with EtOAc. Solvent was removed under vacuum yielding the title as a brown solid (93% yield). ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.89 (d, 2H), 7. 81 (d, 2H), 4.41 (s, 2H), 3.60 (m, 4H), 2.40 (m, 4H).

[00527] p-({4-[3-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)propionyl]-1- piperazinyl}methyl)benzonitrile (268)

[00528] Prepared using General Procedure N, from compound 267 and compound 44b, followed by General Procedure T. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.62 (m, 2H), 7.42 (d, 2H), 7.25 (m, 1 H), 7.12 (d, 2H), 3.50 (d, 6H), 2.95 (m, 4H), 2.21 (m, 4H). ES-MS: 417.0 [M-1], [00529] Preparation of 8-Methoxy-2-[3-(1-{[5-(methylamino)carbonyl-2- pyridyl]carbonyl}-3-azetidinylamino)-3-oxopropyl]-3/7-quinazolin-4-one (273)

[00530] Methyl 6-{[3-(tert-butoxycarbonylamino)-1- azetidinyl]carbonyl}nicotinate (269) [00531] Prepared using General Procedure N, from 5-Methoxycarbonyl-2- pyridinecarboxylic acid and 3-(tert-Butoxycarbonylamino)azetidine. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 9.15 (s, 1 H), 8.39 (dd, 1 H), 8.18 (d, 1 H), 5.10-4.89 (m, 2H), 4.62-4.47 (m, 2H), 4.07-3.97 (m, 1 H), 3.97 (s, 3H), 1.46 (s, 9H).

[00532] Methyl 6-[(3-amino-1-azetidinyl)carbonyl]nicotinate, trifluoroacetate salt (270)

[00533] Prepared using General Procedure D2, from compound 269. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.87 (s, 1 H), 8.19 (dd, 1 H), 7.88 (d, 1 H), 4.92-4.80 (m, 1 H), 4.63-4.52 (m, 1 H), 4.39-4.26 (m, 1 H), 4.08-3.95 (m, 2H), 3.73 (s, 3H).

[00534] Methyl 6-({3-[3-(8-methoxy-4-oxo-3H-quinazolin-2- yl)propionylamino]-1 -azetidinyl}carbonyl)nicotinate (271 )

[00535] Prepared using General Procedure N, from compound 270 and compound 44a. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 9.19-9.06 (m, 1 H), 9.07 (d, 1 H), 8.44 (dd, 1 H), 8.08 (d, 1 H), 7.56 (d, 1 H), 7.29-7.12 (m, 2H), 4.83-4.75 (m, 1 H), 4.59-4.46 (m, 1 H), 4.47-4.28 (m, 2H), 4.00-3.93 (m, 1 H), 3.92 (s, 3H), 3.83 (s, 3H), 2.86-2.75 (m, 2H), 2.65-2.54 (m, 2H). LR ESI MS: m/z calcd. for C23H24N5O6 [M+H]⁺ 466.5, found 466.8.

[00536] 6-({3-[3-(8-Methoxy-4-oxo-3H-quinazolin-2-yl)propionylamino]-1- azetidinyl}carbonyl)nicotinic acid (272)

[00537] Compound 271 (1 equiv.) was solubilized in MeOH. 1 M aq. NaOH solution (2 equiv.) was added, and the mixture was heated at 70 °C for 30 min in microwave. Flash chromatography purification yielded the acid product 272. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 9.05 (br s, 1 H), 8.40 (d, 1 H), 8.06 (d, 1 H), 7.26 (d, 1 H), 7.41-7.22 (m, 2H), 4.87-4.74 (m, 1 H), 4.59-4.26 (m, 3H), 3.99-3.89 (m, 1 H), 3.86 (s, 3H), 2.94-2.79 (m, 2H), 2.70-2.57 (m, 2H). LR ESI MS: m/z calcd. for C22H22N5O6 [M+H]⁺ 452.4, found 452.8.

[00538] 8-Methoxy-2-[3-(1-{[5-(methylamino)carbonyl-2-pyridyl]carbonyl}- 3-azetidinylamino)-3-oxopropyl]-3/7-quinazolin-4-one (273)

[00539] Prepared using General Procedure N, from compound 272 and Methylamine. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 9.19 (d, 1 H), 9.08 (s, 1 H), 8.44 (dd, 1 H), 8.08 (d, 1 H), 7.57 (d, 1 H), 7.27-7.12 (m, 2H), 4.85-4.75 (m, 1 H), 4.60-4.47 (m, 1 H), 4.47-4.28 (m, 2H), 3.99-3.92 (m, 1 H), 3.92 (s, 3H), 3.83 (s, 3H), 2.85-2.76 (m, 2H), 2.64-2.54 (m, 2H). LR ESI MS: m/z calcd. for C23H23N6O5 [M-Hp 463.5, found 463.9.

[00540] Preparation of 6-{(/?)-3-[3-(8-Methoxy-4-oxo-3H-quinazolin-2- yl)propionylamino]-1 -pyrrolidinyl}nicotinonitrile (276)

[00541 ] 2-[(R)-3-(tert-Butoxycarbonylamino)-1 -pyrrolidinyl]-5- pyridinecarbonitrile (274)

[00542] Prepared using General Procedure W, from (R)-3-(tert- Butoxycarbonylamino)pyrrolidine and 6-Chloronicotinonitrile. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.41-8.35 (m, 1 H), 7.60-7.52 (m, 1 H), 6.73-6.62 (m, 1 H), 4.82-4.68 (m, 1 H), 4.41-4.25 (m, 1 H), 3.44-3.31 (m, 2H), 2.36-2.21 (m, 1 H), 2.07- 1 .91 (m, 1 H), 1 .43 (m, 9H). ESI MS: m/z calcd for C15H20N4O2 [M+H]⁺, 289.34.

[00543] (/?)-6-(3-Aminopyrrolidin-1-yl)nicotinonitrile, hydrochloride salt (275) [00544] Prepared using General Procedure D, from compound 274. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.67-8.50 (m, 1 H), 8.21-8.04 (m, 1 H), 7.35-7.09 (m, 1 H), 4.36-3.55 (m, 6H), 2.76-2.52 (m, 1 H), 2.50-2.30 (m, 1 H). ESI MS: m/z calcd for C10H13CIN4 [M+H]⁺, 189.22. [00545] 6-{(/?)-3-[3-(8-Methoxy-4-oxo-3H-quinazolin-2-yl)propionylamino]-

1 -pyrrolidinyl}nicotinonitrile (276)

[00546] Prepared using General Procedure N, from compound 275 and compound 44a. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 11.42 (s, 1 H), 8.36-8.28 (m, 1 H), 7.85-7.76 (m, 1 H), 7.54-7.46 (m, 1 H), 7.43-7.32 (m, 1 H), 7.18-7.10 (m, 1 H),

7.03-6.91 (m, 1 H), 6.27-6.18 (m, 1 H), 4.69-4.55 (m, 1 H), 3.94 (s, 3H), 3.80-3.67 (m, 1 H), 3.61-3.22 (m, 3H), 3.17-3.06 (m, 1 H), 2.75-2.72 (m, 2H), 2.37-2.20 (m, 1 H), 2.11-1.95 (m, 1 H). LR ESI MS: m/z calcd for C22H22N6O3 [M+H]⁺, 419.44.

[00547] Preparation of A/2-(1-(3-(8-Methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)pyrrolidin-3-yl)-A/5-methylpyridine-2,5-dicarboxamide (281 )

[00548] Methyl 6-(1 -(tert-butoxycarbonyl)pyrrolidin-3- ylcarbamoyl)nicotinate (277) [00549] Prepared using General Procedure N, from 5-Methoxycarbonyl-2- pyridinecarboxylic acid and ferf-Butyl 3-amino-1-pyrrolidinecarboxylate. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 9.21 (s, 1 H), 8.48-8.40 (m, 1 H), 8.30-8.21 (m, 1 H), 8.18-8.06 (m, 1 H), 4.72-4.58 (m, 1 H), 3.98 (s, 3H), 3.79-3.67 (m, 1 H), 3.63-3.25 (m, 3H), 2.33-2.18 (m, 1 H), 2.09-1.90 (m, 1 H), 1.46 (s, 9H). LR ESI MS: m/z calcd for C17H23N3O5 [M+H]⁺, 350.38.

[00550] Methyl 6-(pyrrolidin-3-ylcarbamoyl)nicotinate, hydrochloride salt (278)

[00551] Prepared using General Procedure D, from compound 277. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 9.46-9.25 (m, 2H), 9.20-9.05 (m, 1 H), 8.54-8.44 (m, 1 H), 8.22-8.14 (m, 1 H), 4.70-4.55 (m, 1 H), 3.93 (s, 3H), 3.47-3.29 (m, 2H), 3.29-3.16 (m, 2H), 2.30-2.14 (m, 1 H), 2.11-1.95 (m, 1 H). LR ESI MS: m/z calcd for C12H16CIN3O3 [M+H]⁺, 250.26.

[00552] Methyl 4-(1 -(3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)pyrrolidin-3-ylcarbamoyl)benzoate (279)

[00553] Prepared using General Procedure N, from compound 278 and compound 44a. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 9.09-9.00 (m, 1 H), 8.47- 8.37 (m, 1 H), 8.28-8.09 (m, 2H), 7.86-7.77 (m, 1 H), 7.39-7.29 (m, 1 H), 7.19-7.08 (m, 1 H), 4.79-4.60 (m, 1 H), 3.97 (s, 6H), 3.92-3.82 (m, 1 H), 3.82-3.56 (m, 2H), 3.55-3.45 (m, 1 H), 3.23-3.08 (m, 2H), 2.89-2.73 (m, 2H), 2.45-1.97 (m, 2H). LR ESI MS: m/z calcd for C25H26N4O6 [M+H]⁺, 479.49.

[00554] 4-(1 -(3-(8-Methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)pyrrolidin-3-ylcarbamoyl)benzoic acid (280)

[00555] Compound 279 (1 equiv.) was solubilized in MeOH. 1 M aq. NaOH solution (2 equiv.) was added, and the mixture was heated at 70 °C for 30 min. Flash chromatography purification yielded the acid product 280. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 9.01-8.87 (m, 1 H), 8.41-8.30 (m, 1 H), 8.09-7.98 (m, 1 H), 7.63-7.55 (m, 1 H), 7.32-7.22 (m, 1 H), 7.21-7.13 (m, 1 H), 4.66-4.44 (m, 2H), 4.01- 3.90 (m, 1 H), 3.89-3.80 (m, 3H), 3.76-3.59 (m, 2H), 3.59-3.36 (m, 2H), 2.99-2.88 (m, 2H), 2.87-2.74 (m, 2H), 2.35-1.92 (m, 2H). LR ESI MS: m/z calcd for C24H24N4O6 [M- H]-, 463.47.

[00556] A/2-(1 -(3-(8-Methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)pyrrolidin-3-yl)-/V5-methylpyridine-2,5-dicarboxamide (281 )

[00557] Prepared using General Procedure N, from compound 280 and Methylamine. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.91-8.68 (m, 1 H), 8.28- 8.09 (m, 1 H), 8.09-7.91 (m, 1 H), 7.68-7.49 (m, 1 H), 7.32-7.05 (m, 2H), 4.66-4.40 (m, 1 H), 4.04-3.75 (m, 4H), 3.76-3.31 (m, 3H), 3.05-2.65 (m, 7H), 2.43-1 .85 (m, 2H). LR ESI MS: m/z calcd for C24H26N6O5 [M+H]⁺, 479.50.

[00558] Preparation of Methyl 6-(1-(3-(8-hydroxy-4-oxo-3,4- dihydroquinazolin-2-yl)propanoyl)pyrrolidin-3-ylcarbamoyl)nicotinate (283) and A/2-(1-(3-(8-Hydroxy-4-oxo-3,4-dihydroquinazolin-2-yl)propanoyl)pyrrolidin-3- yl)-/V5-methylpyridine-2,5-dicarboxamide (286)

[00559] Methyl 6-(1 -(3-(8-(benzyloxy)-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)pyrrolidin-3-ylcarbamoyl)nicotinate (282)

[00560] Prepared using General Procedure N, from compound 278 and compound 44b. ¹H NMR (300 MHz, DMSO-d6): 6 (ppm) 12.22 (s, 1 H), 9.14-8.97 (m, 2H), 8.53-8.42 (m, 1 H), 8.22-8.11 (m, 1 H), 7.70-7.60 (m, 1 H), 7.55-7.43 (m, 2H), 7.43-7.27 (m, 5H), 5.24 (s, 2H), 4.60-4.34 (m, 1 H), 3.92 (s, 3H), 3.85-3.45 (m, 4H), 2.96-2.74 (m, 4H), 2.15-1.92 (m, 2H). LR ESI MS: m/z calcd for C₃oH29N₅06[M+H]⁺, 556.58. [00561] Methyl 6-(1-(3-(8-hydroxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)pyrrolidin-3-ylcarbamoyl)nicotinate (283)

[00562] Prepared using General Procedure T, from compound 282. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.56-7.35 (m, 2H), 7.29-7.14 (m, 2H), 7.13-7.02 (m, 1 H), 3.84-3.72 (m, 1 H), 3.70-3.55 (m, 1 H), 3.51 (s, 3H), 3.46-3.30 (m, 1 H), 3.22-

3.16 (m, 2H), 2.96-2.75 (m, 4H), 2.32-1.60 (m, 6H). LR ESI MS: m/z calcd for C23H23N₅O6[M+H]⁺, 446.45.

[00563] 6-(1 -(3-(8-(Benzyloxy)-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)pyrrolidin-3-ylcarbamoyl)nicotinic acid (284) [00564] Compound 282 (1 equiv.) was solubilized in MeOH. 1 M aq. NaOH solution (2 equiv.) was added, and the mixture was heated at 70 °C for 30 min. Flash chromatography purification yielded the acid product 284. ¹H NMR (300 MHz, DMSO- d6) 5 (ppm) 9.07 (s, 1 H), 8.50-8.41 (m, 1 H), 8.19-8.10 (m, 1 H), 7.75-7.64 (m, 1 H), 7.58-7.19 (m, 7H), 5.33 (s, 2H), 3.86-3.42 (m, 5H), 3.41-3.27 (m, 2H), 3.12-2.99 (m, 2H), 2.94-2.80 (m, 2H), 2.22-2.00 (m, 2H). LR ESI MS: m/z calcd for C29H27N5O6 [M- H]-, 540.55.

[00565] A/2-(1 -(3-(8-(Benzyloxy)-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)pyrrolidin-3-yl)-/V5-methylpyridine-2,5-dicarboxamide (285)

[00566] Prepared using General Procedure N, from compound 284 and Methylamine. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.23 (s, 1 H), 9.11-8.88 (m, 1 H), 8.86-8.67 (m, 1 H), 8.43-8.25 (m, 1 H), 7.74-7.58 (m, 1 H), 7.57-7.23 (m, 5H), 5.26 (s, 2H), 4.62-4.31 (m, 1 H), 3.91-3.40 (m, 2H), 2.98-2.67 (m, 5H), 2.15-1.89 (m, 2H). LR ESI MS: m/z calcd for C30H30N6O5 [M+H]⁺, 555.55.

[00567] A/2-(1 -(3-(8-Hydroxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)pyrrolidin-3-yl)-/V5-methylpyridine-2,5-dicarboxamide (286)

[00568] Prepared using General Procedure T, from compound 285. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.16 (s, 1 H), 9.42-8.65 (m, 4H), 8.48-7.98 (m, 2H), 7.49 (s, 1 H), 7.38-7.00 (m, 2H), 4.73-4.35 (m, 1 H), 4.02-3.43 (m, 4H), 3.11-2.64 (m, 8H), 2.34-1.89 (m, 2H), 1.43-1.03 (m, 1 H). LR ESI MS: m/z calcd for C23H24N6O5 [M+H]⁺, 465.47.

[00569] Preparation of (R)-Methyl 6-(1 -(3-(8-methoxy-4-oxo-3,4- dihydroquinazolin-2-yl)propanoyl)pyrrolidin-3-ylcarbamoyl)nicotinate (289) and (S)-Methyl 6-(1 -(3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)pyrrolidin-3-ylcarbamoyl)nicotinate (292)

[00570] (R)-tert-Butyl 1 -(3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)pyrrolidin-3-ylcarbamate (287)

[00571] Prepared using General Procedure N, from compound 44a and (R)-3- (ferf-Butoxycarbonylamino)pyrrolidine. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.61-7.53 (m, 1 H), 7.31-7.22 (m, 1 H), 7.22-7.14 (m, 1 H), 4.13-3.92 (m, 2H), 3.86 (s, 3H), 3.77-3.27 (m, 4H), 3.23-3.18 (m, 1 H), 2.95-2.85 (m, 2H), 2.80-2.69 (m, 2H), 2.17-1.65 (m, 2H), 1.39-1.23 (m, 9H). ESI MS: m/z calcd for C21H28N4O5 [M+H]⁺, 417.47.

[00572] (R)-2-(3-(3-Aminopyrrolidin-1 -yl)-3-oxopropyl)-8- methoxyquinazolin-4(3/7)-one, hydrochloride salt (288)

[00573] Prepared using General Procedure D, from compound 287. ¹H NMR (300 MHz, Methanol-cM): 6 (ppm) 7.92-7.79 (m, 1 H), 7.76-7.56 (m, 2H), 4.14 (s, 3H), 3.47-3.26 (m, 4H), 3.24-3.04 (m, 2H), 2.58-2.00 (m, 2H). ESI MS: m/z calcd for C16H21CIN4O3 [M+H]⁺, 317.35.

[00574] (R)-Methyl 6-(1 -(3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)pyrrolidin-3-ylcarbamoyl)nicotinate (289)

[00575] Prepared using General Procedure N, from compound 288 and 5- Methoxycarbonyl-2-pyridinecarboxylic acid. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 11.53 (s, 1 H), 9.21-9.10 (m, 1 H), 8.58-8.46 (m, 1 H), 8.39-8.20 (m, 2H), 7.98-7.86 (m, 1 H), 7.51-7.39 (m, 1 H), 7.29-7.19 (m, 1 H), 4.90-4.70 (m, 1 H), 4.07 (s, 6H), 4.03- 3.56 (m, 4H), 3.33-3.20 (m, 2H), 3.02-2.82 (m, 2H), 2.57-1.95 (m, 4H). LR ESI MS: m/z calcd for C24H25N5O6 [M+H]⁺, 480.48.

[00576] (S)-tert-Butyl 1 -(3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)pyrrolidin-3-ylcarbamate (290)

[00577] Prepared using General Procedure N, from compound 44a and (S)-3- (ferf-Butoxycarbonylamino)pyrrolidine. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.83-7.62 (m, 1 H), 7.52-7.29 (m, 2H), 3.99 (s, 3H), 3.83-3.38 (m, 4H), 3.10-2.96 (m, 2H), 2.96-2.76 (m, 2H), 2.41-1.70 (m, 2H), 1.59-1.36 (m, 9H). ESI MS: m/z calcd for C21H28N4O5 [M+H]⁺, 417.47.

[00578] (S)-2-(3-(3-Aminopyrrolidin-1 -yl)-3-oxopropyl)-8- methoxyquinazolin-4(3/7)-one, hydrochloride salt (291)

[00579] Prepared using General Procedure D, from compound 290. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.87-7.76 (m, 1 H), 7.75-7.53 (m, 2H), 4.12 (s, 3H), 3.93-3.51 (m, 4H), 3.48-3.25 (m, 4H), 3.22-3.06 (m, 2H), 2.16-2.01 (m, 2H). ESI MS: m/z calcd for C16H21CIN4O3 [M+H]⁺, 317.35.

[00580] (S)-Methyl 6-(1 -(3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)pyrrolidin-3-ylcarbamoyl)nicotinate (292)

[00581] Prepared using General Procedure N, from compound 291 and 5- Methoxycarbonyl-2-pyridinecarboxylic acid. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 11.98 (s, 1 H), 9.13-9.02 (m, 1 H), 8.57-8.41 (m, 1 H), 7.95-7.83 (m, 1 H), 7.50- 7.38 (m, 1 H), 7.29-7.19 (m, 1 H), 4.97-4.73 (m, 1 H), 4.14-4.03 (m, 6H), 4.03-3.72 (m, 3H), 3.58 (s, 2H), 3.38-3.22 (m, 2H), 3.15-2.93 (m, 1 H), 2.61-2.12 (m, 2H). LR ESI MS: m/z calcd for C24H25N5O6 [M+H]⁺, 480.48.

[00582] Preparation of A/2-(1-(3-(8-Mmethoxy-4-oxo-3,4-dihydroquinazolin- 2-yl)propanoyl)piperidin-4-yl)-/V5-methylpyridine-2,5-dicarboxamide (297)

[00583] Methyl 6-(1-(tert-butoxycarbonyl)piperidin-4- ylcarbamoyl)nicotinate (293)

[00584] Prepared using General Procedure N, from 5-Methoxycarbonyl-2- pyridinecarboxylic acid and te/Y-Butyl 4-amino-1-piperidinecarboxylate. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.79-7.72 (m, 1 H), 7.49-7.40 (m, 1 H), 7.40-7.34 (m, 1 H), 4.43-4.29 (m, 1 H), 4.07-3.95 (m, 4H), 3.68-3.53 (m, 1 H), 3.30-3.16 (m, 1 H), 3.00 (s, 4H), 2.93-2.77 (m, 1 H), 2.05-1.79 (m, 2H), 1.46 (s, 9H), 1.42-1.22 (m, 1 H). ESI MS: m/z calcd for C18H24N3O5 [M+H]⁺, 364.40. [00585] Methyl 6-(piperidin-4-ylcarbamoyl)nicotinate, hydrochloride salt

(294)

[00586] Prepared using General Procedure D, from compound 293. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.89-7.81 (m, 1 H), 7.76-7.61 (m, 2H), 4.67-4.53 (m, 1 H), 4.21-4.00 (m, 4H), 3.28-3.09 (m, 2H), 2.79-2.71 (m, 1 H), 2.22-1 .98 (m, 2H), 1 .79-1 .43 (m, 2H). ESI MS: m/z calcd for C13H18CIN3O3 [M+H]⁺, 264.29.

[00587] Methyl 6-(1 -(3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)piperidin-4-ylcarbamoyl)nicotinate (295)

[00588] Prepared using General Procedure N, from compound 294 and compound 44a. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 9.09 (s, 1 H), 8.47-8.39 (m, 1 H), 8.28-8.21 (m, 1 H), 8.07-7.98 (m, 1 H), 7.86-7.78 (m, 1 H), 7.44-7.31 (m, 2H), 7.19-7.11 (m, 1 H), 4.28-4.09 (m, 1 H), 3.97 (s, 3H), 3.90 (s, 3H), 3.31-3.10 (m, 3H), 2.99-2.80 (m, 3H), 2.19-1.97 (m, 2H), 1.64-1.44 (m, 2H). ESI MS: m/z calcd for C25H27N5O6 [M+H]⁺, 494.51.

[00589] Sodium 6-(1 -(3-(8-methoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)piperidin-4-ylcarbamoyl)nicotinate (296)

[00590] Compound 295 (1 equiv.) was solubilized in MeOH. 1 M aq. NaOH solution (2 equiv.) was added, and the mixture was heated at 70 °C for 30 min. Concentration to dryness yielded the title compound. ¹H NMR (300 MHz, Methanol- d4) 5 (ppm) 9.13 (s, 1 H), 8.45-8.34 (m, 1 H), 8.14-8.03 (m, 1 H), 7.79-7.69 (m, 1 H), 7.30-7.19 (m, 1 H), 7.18-7.08 (m, 1 H), 4.62-4.48 (m, 1 H), 4.21-4.05 (m, 2H), 3.99 (s, 3H), 3.30-3.15 (m, 1 H), 3.08-2.75 (m, 5H), 2.05-1 .89 (m, 2H), 1 .64-1 .43 (m, 2H). ESI MS: m/z calcd for C24H₂4N₅NaO6 [M+H]⁺, 502.16.

[00591] A/2-(1-(3-(8-Mmethoxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)piperidin-4-yl)-/V5-methylpyridine-2,5-dicarboxamide (297)

[00592] Prepared using General Procedure N, from compound 296 and Methylamine. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 11.21 (s, 1), 8.91 (s, 1 H), 8.21 (s, 2H), 8.02-7.92 (m, 1 H), 7.87-7.79 (m, 1 H), 7.43-7.32 (m, 1 H), 7.22-7.14 (m, 1 H), 6.51 (s, 1 H), 4.68-4.50 (m, 1 H), 4.29-4.08 (m, 1 H), 4.00 (s, 3H), 3.90-3.75 (m, 1 H), 3.31-3.09 (m, 3H), 3.08-2.98 (m, 3H), 2.98-2.75 (m, 3H), 2.19-1.94 (m, 2H), 1.87-1.40 (m, 3H), 0.96-0.72 (m, 1 H). ESI MS: m/z calcd for C25H28N6O5 [M+H]⁺, 493.52.

[00593] Preparation of 2-(3-{(1/?,5S)-7-[5-(Methylamino)carbonyl-2-pyridyl]- 3,7-diazabicyclo[3.3.0]oct-3-yl}-3-oxopropyl)-8-hydroxy-3/7-quinazolin-4-one (302)

[00594] General Procedure X

[00595] To a stirred solution of methyl ester intermediate (298, 500 mg) in 20 mL of MeOH was added 20 mL of Methylamine solution (40 wt. % in H2O) and stirred for 48 hours at 40 °C. The reaction mixture was then cooled to ambient temperature and the solvent was removed by rotary evaporation. The crude residue was diluted with EtOAc and washed with water, brine and dried over anhydrous Na2SO4 and concentrated to obtain crude material which was purified by flash column chromatography by using 0 to 5% of MeOH in DCM to afford the white solid of methyl amide intermediate (299).

[00596] tert-Butyl (1/?,5S)-7-(5-methoxycarbonyl-2-pyridyl)-3,7- diazabicyclo[3.3.0]octane-3-carboxylate (298)

[00597] Prepared using General Procedure W, from tert-Butyl (1 R,5S)-3,7- diazabicyclo[3.3.0]octane-3-carboxylate and Methyl 6-chloronicotinate. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.82-8.76 (m, 1 H), 8.04-7.97 (m, 1 H), 6.35-6.28 (m, 1 H), 3.86 (s, 3H), 3.82-3.60 (m, 4H), 3.52-3.22 (m, 4H), 3.08-2.97 (m, 2H), 1.45 (s, 9H). LR ESI MS: m/z calcd for C18H27N3O4 [M+H]⁺, 348.8 found 348.4.

[00598] tert-Butyl (1 /?,5S)-7-[5-(methylamino)carbonyl-2-pyridyl]-3,7- diazabicyclo[3.3.0]octane-3-carboxylate (299) Ill

[00599] Prepared using General Procedure X, from compound 298. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.56-8.50 (m, 1 H), 7.94-7.86 (m, 1 H), 6.38-6.30 (m, 1 H), 6.00 (br s, 1 H), 3.80-3.59 (m, 4H), 3.51-3.21 (m, 4H), 3.06-2.95 (m, 4H), 2.66-2.47 (m, 2H), 1.45 (s, 9H). LR ESI MS: m/z calcd for C18H27N4O3 [M+H]⁺, 347.4 found 347.8.

[00600] (6-{( 1 R,5S)-3,7-Diazabicyclo[3.3.0]oct-3-yl}-3- pyridyl)(methylamino)formaldehyde, hydrochloride salt (300)

[00601] Prepared using General Procedure D, from compound 299. ¹H NMR (300 MHz, D2O): 5 (ppm) 8.26-8.22 (m, 1 H), 8.19-8.13 (m, 1 H), 7.07-7.00 (m, 1 H), 3.98-3.89 (m, 2H), 3.69-3.59 (m, 4H), 3.43-3.45 (m, 2H), 3.31-3.22 (m, 2H), 2.83 (s, 3H). LR ESI MS: m/z calcd for C13H19N4O [M+H]⁺, 247.3 found 247.8.

[00602] 2-(3-{(1R,5S)-7-[5-(Methylamino)carbonyl-2-pyridyl]-3,7- diazabicyclo[3.3.0]oct-3-yl}-3-oxopropyl)-8-(benzyloxy)-3/7-quinazolin-4-one (301 )

[00603] Prepared using General Procedure N, from compound 300 and compound 44b. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.35-12.02 (br s, 1 H), 8.67- 8.42 (m, 1 H), 8.26-7.78 (m, 2H), 7.73-7.12 (m, 7H), 6.33-6.20 (m, 1 H), 5.21 (s, 2H), 3.84-2.23 (m, 17H). LR ESI MS: m/z calcd for C31H33N6O4 [M+H]⁺, 553.6 found 553.9.

[00604] 2-(3-{(1R,5S)-7-[5-(Methylamino)carbonyl-2-pyridyl]-3,7- diazabicyclo[3.3.0]oct-3-yl}-3-oxopropyl)-8-hydroxy-3/7-quinazolin-4-one (302)

[00605] Prepared using General Procedure T, from compound 301. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.13 (br s, 1 H), 9.20 (br s, 1 H), 8.59-8.53 (m, 1 H), 8.20-8.1 1 (m, 1 H), 7.94-7.86 (m, 1 H), 7.53-7.45 (m, 1 H), 7.30-7.22 (m, 1 H), 7.19- 7.12 (m, 1 H), 6.48-6.39 (m, 1 H), 3.90-2.82 (m, 14H), 2.77-2.72 (m, 3H). LR ESI MS: m/z calcd for C24H27N6O7 [M+H]⁺, 463.5 found 463.9. [00606] Preparation of 2-(3-{(1/?,5S)-7-[p-(Methylamino)carbonylphenyl]- 3,7-diazabicyclo[3.3.0]oct-3-yl}-3-oxopropyl)-8-hydroxy-3/7-quinazolin-4-one (307)

[00607] tert-Butyl (1/?,5S)-7-(p-methoxycarbonylphenyl)-3,7- diazabicyclo[3.3.0]octane-3-carboxylate (303)

To a stirred solution of Methyl p-bromobenzoate (1 equiv.) in anhydrous toluene was added tert-Butyl (1R,5S)-3,7-diazabicyclo[3.3.0]octane-3-carboxylate (1.8 equiv.), XantPhos (10 mol%), Pd(dba)3 (10 mol %), and CS2CO3 (3 equiv.) at ambient temperature. The resulting suspension was heated to 100 °C and stirred for 16 hours. The reaction mixture was then cooled to ambient temperature and diluted with EtOAc and filtered. The filtrate was concentrated to obtain crude material which was purified by flash column chromatography by using 0 to 30% of ethyl acetate in hexane to afford the white solid of ester intermediate 303. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.97-7.78 (m, 2H), 6.57-6.37 (m, 2H), 3.84 (s, 3H), 3.62-3.26 (m, 8H), 3.00 (s, 2H), 1.45 (s, 9H).

[00608] tert-Butyl (1 R,5S)-7-[p-(methylamino)carbonylphenyl]-3,7- diazabicyclo[3.3.0]octane-3-carboxylate (304)

Prepared using General Procedure X, from compound 303. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.69-7.63 (m, 2H), 6.54-6.47 (m, 2H), 5.97 (bs, 1 H), 3.70- 3.51 (m, 4H), 3.41-3.19 (m, 4H), 3.05-2.90 (m, 5H), 1.45 (s, 9H).

[00609] (P-{(1 R,5S)-3,7-Diazabicyclo[3.3.0]oct-3- yl}phenyl)(methylamino)formaldehyde, hydrochloride salt (305) [00610] Prepared using General Procedure D, from compound 304. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.70-7.60 (m, 2H), 6.77-6.69 (m, 2H), 3.61-3.35 (m, 6H), 3.25-3.13 (m, 4H), 2.83 (s, 3H).

[00611 ] 2-( 3-{( 1 R,5S)-7-[p-(Methylamino)carbonylphenyl]-3,7- diazabicyclo[3.3.0]oct-3-yl}-3-oxopropyl)-8-(benzyloxy)-3/7-quinazolin-4-one (306)

[00612] Prepared using General Procedure N, from compound 305 and compound 44b. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.22 (bs, 1 H), 8.59-8.53 (m, 1 H), 8.20-8.11 (m, 1 H), 7.94-7.86 (m, 1 H), 7.53-7.45 (m, 1 H), 7.45-7.27 (m, 7H), 6.46-6.38 (m, 1 H), 5.24 (s, 2H), 3.78-3.35 (m, 13H), 2.86-2.74 (m, 2H), 2.84 (s, 2H). LR ESI MS: m/z calcd for C32H33N5O4 : 551 .63; found [M+H]⁺ , found 552.8.

[00613] 2-(3-{( 1 R,5S)-7-[p-(Methylamino)carbonylphenyl]-3,7- diazabicyclo[3.3.0]oct-3-yl}-3-oxopropyl)-8-hydroxy-3/7-quinazolin-4-one (307)

[00614] Prepared using General Procedure T, from compound 306. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 8.02-7.96 (bs, 1 H), 7.70-7.59 (m, 3H), 7.34-7.27 (m, 1 H), 7.23-7.17 (m, 1 H), 6.51-6.43 (m, 2H), 3.85-3.75 (m, 2H), 3.69-3.50 (m, 3H), 3.29- 2.96 (m, 8H), 2.86 (s, 2H), 2.85-2.75 (m, 2H). LR ESI MS: m/z calcd for C25H27N5O4 : 461.51 ; found [M+H]+ : 460.80.

[00615] Preparation of 6-{( 1 R,5S)-7-[3-(8-Hydroxy-4-oxo-3H-quinazolin-2- yl)propionyl]-3,7-diazabicyclo[3.3.0]oct-3-yl}nicotinonitrile (311 )

[00616] tert-Butyl (1/?,5S)-7-(5-cyano-2-pyridyl)-3,7- diazabicyclo[3.3.0]octane-3-carboxylate (308)

[00617] Prepared using General Procedure W, from ferf-Butyl (1 R,5S)-3,7- diazabicyclo[3.3.0]octane-3-carboxylate and 6-Bromonicotinonitrile. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.44-8.36 (m, 1 H), 7.64-7.55 (m, 1 H), 6.37-6.29 (m, 1 H), 3.82-3.21 (m, 8H), 3.09-2.98 (m, 2H), 1.45 (s, 9H). LR ESI MS: m/z calcd for C17H23N4O2 [M+H]⁺, 315.4 found 315.6.

[00618] 6-{(1 R,5S)-3,7-Diazabicyclo[3.3.0]oct-3-yl}nicotinonitrile, hydrochloride salt (309)

[00619] Prepared using General Procedure D, from compound 308. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.58-8.53 (m, 1 H), 8.14-8.07 (m, 1 H), 7.18-7.10 (m, 1 H), 4.03-3.93 (m, 2H), 3.85-3.77 (m, 2H), 3.72-3.61 (m, 2H), 3.49-3.32 (m, 4H). LR ESI MS: m/z calcd for C12H14N4 [M+H]⁺, 215.3 found 215.4.

[00620] 6-[(1/?,5S)-7-{3-[8-(Benzyloxy)-4-oxo-3H-quinazolin-2-yl]propionyl}- 3,7-diazabicyclo[3.3.0]oct-3-yl]nicotinonitrile (310)

[00621] Prepared using General Procedure N, from compound 309 and compound 44b. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.27-12.13 (br s, 1 H), 7.84- 7.75 (m, 1 H), 7.69-7.61 (m, 1 H), 7.53-7.24 (m, 7H), 6.49-6.38 (m, 1 H), 5.24 (s, 2H), 3.82-3.13 (m, 8H), 3.00-2.71 (m, 6H). LR ESI MS: m/z calcd for C30H29N6O3 [M+H]⁺ , 521.6 found 521.8.

[00622] 6-{(1/?,5S)-7-[3-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)propionyl]- 3,7-diazabicyclo[3.3.0]oct-3-yl}nicotinonitrile (311 )

[00623] Prepared using General Procedure T, from compound 310. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.20-12.07 (br s, 1 H), 9.31-9.15 (m, 1 H), 8.51-8.43 (m, 1 H), 7.87-7.77 (m, 1 H), 7.53-7.44 (m, 1 H), 7.30-7.21 (m, 1 H), 7.19-7.12 (m, 1 H), 6.60-6.47 (m, 1 H), 3.90-3.47 (m, 5H), 3.41-3.07 (m, 5H), 2.95-2.80 (m, 4H). LR ESI MS: m/z calcd for C30H29N6O3 [M-H]’ , 429.5 found 429.8.

[00624] Preparation of 8-Hydroxy-2-(3-{6-[5-(methylamino)carbonyl-2- pyridyl]-2,6-diaza-2-spiro[3.3]heptyl}-3-oxopropyl)-3/7-quinazolin-4-one (315)

[00625] tert-Butyl 6-(5-methoxycarbonyl-2-pyridyl)-2,6-diaza-2- spiro[3.3]heptanecarboxylate (312)

[00626] Prepared using General Procedure W, from tert-Butyl 2,6-diaza-2- spiro[3.3]heptanecarboxylate and Methyl 6-chloronicotinate. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.82-8.72 (m, 1 H), 8.04-7.95 (m, 1 H), 6.28-6.17 (m, 1 H), 4.22 (s, 4H), 4.12 (s, 4H), 3.86 (s, 3H), 1.45 (s, 9H). LR ESI MS: m/z calcd for C17H24N3O4 [M+H]⁺, 334.4 found 334.8.

[00627] tert-Butyl 6-[5-(methylamino)carbonyl-2-pyridyl]-2,6-diaza-2- spiro[3.3]heptanecarboxylate (313)

[00628] Prepared using General Procedure X, from compound 312. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.53-8.47 (m, 1 H), 7.93-7.85 (m, 1 H), 6.29-6.23 (m, 1 H), 5.95 (br s, 1 H), 4.19 (s, 4H), 4.12 (s, 4H), 3.01-2.96 (m, 3H), 1.45 (s, 9H). LR ESI MS: m/z calcd for C17H25N4O3 [M+H]⁺, 333.4 found 333.8.

[00629] 8-(Benzyloxy)-2-(3-{6-[5-(methylamino)carbonyl-2-pyridyl]-2,6- diaza-2-spiro[3.3]heptyl}-3-oxopropyl)-3/7-quinazolin-4-one (314)

[00630] Compound 313 was treated according to General Procedure D, to give (cfe-eoc)-deprotected amine intermediate as hydrochloride salt (LR ESI MS: m/z calcd for C12H17N4O [M+H]⁺, 233.4 found 233.7). The deprotected amine was used as such according to General Procedure N, with compound 44b. ¹H NMR (300 MHz, DMSO- d6) 5 (ppm) 12.34-12.09 (br s, 1 H), 8.62-8.42 (m, 1 H), 8.33-8.09 (m, 1 H), 7.99-7.79 (m, 1 H), 7.76-7.13 (m, 7H), 6.46-6.21 (m, 1 H), 5.25 (s, 2H), 4.53-3.78 (m, 8H), 3.57- 2.05 (m, 7H). LR ESI MS: m/z calcd for C30H31N6O4 [M+H]⁺, 539.6 found 539.9.

[00631] 8-Hydroxy-2-(3-{6-[5-(methylamino)carbonyl-2-pyridyl]-2,6-diaza-2- spiro[3.3]heptyl}-3-oxopropyl)-3/7-quinazolin-4-one (315)

[00632] Prepared using General Procedure T, from compound 314. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 8.58-8.51 (m, 1 H), 8.26-8.16 (m, 1 H), 7.97-7.88 (m, 1 H), 7.51-7.43 (m, 1 H), 7.28-7.11 (m, 2H), 6.42-6.34 (m, 1 H), 4.45 (s, 2H), 4.15 (s, 4H), 4.04 (s, 2H), 2.87-2.79 (m, 2H), 2.77-2.71 (m, 3H), 2.67-2.59 (m, 2H). LR ESI MS: m/z calcd for C23H25N6O4 [M+H]⁺ , 449.5 found 449.8.

[00633] Preparation of 6-(5-(3-(8-Hydroxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)-2,5-diazabicyclo[2.2.2]octan-2-yl)-/V-methylnicotinamide (320)

[00634] tert-Butyl 5-(5-methoxycarbonyl-2-pyridyl)-2,5- diazabicyclo[2.2.2]octane-2 -carboxylate (316)

[00635] Prepared using General Procedure W, from te/Y-Butyl 2,5- diazabicyclo[2.2.2]octane-2-carboxylate and Methyl 6-chloronicotinate. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.79-8.74 (m, 1 H), 8.06-7.99 (m, 1 H), 6.35-6.28 (m, 1 H), 3.86 (s, 3H), 3.73-3.41 (m, 5H), 2.17-1.53 (m, 5H), 1.46 (s, 9H). LR ESI MS: m/z calcd for C18H26N3O4 [M+H]⁺, 348.4 found 348.8.

[00636] tert-Butyl 5-[5-(methylamino)carbonyl-2-pyridyl]-2,5- diazabicyclo[2.2.2]octane-2 -carboxylate (317)

[00637] Prepared using General Procedure X, from compound 316. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 8.55-8.45 (m, 1 H), 7.95-7.85 (m, 1 H), 6.40-6.29 (m, 1 H), 5.99-5.86 (br s, 1 H), 3.74-3.39 (m, 5H), 3.02-2.96 (m, 3H), 2.17-1.74 (m, 5H), 1 .46 (s, 9H). LR ESI MS: m/z calcd for C18H27N4O3 [M+H]⁺, 347.4 found 347.8.

[00638] 6-(2,5-Diazabicyclo[2.2.2]octan-2-yl)-A/-methylnicotinamide, hydrochloride salt (318)

[00639] Prepared using General Procedure D, from compound 317. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.54-8.40 (m, 1 H), 7.53-7.36 (m, 1 H), 4.84-4.74 (m, 1 H), 4.27-4.10 (m, 2H), 4.04-3.90 (m, 1 H), 3.81-3.57 (m, 9H), 3.03 (s, 1 H), 2.95 (s, 2H), 2.90 (s, 1 H), 2.43-2.22 (m, 2H), 2.22-2.04 (m, 2H). ESI MS: m/z calcd for C13H19CIN4O1 [M+H]⁺, 247.30.

[00640] 6-(5-(3-(8-(Benzyloxy)-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)-2,5-diazabicyclo[2.2.2]octan-2-yl)-A/-methylnicotinamide (319)

[00641] Prepared using General Procedure N, from compound 318 and compound 44b. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.21 (s, 1 H), 8.65-8.45 (m, 1 H), 8.29-8.05 (m, 1 H), 8.00-7.81 (m, 1 H), 7.74-7.57 (m, 1 H), 7.56-7.15 (m, 6H), 6.62-6.30 (m, 1 H), 5.37-5.10 (m, 2H), 5.03-4.73 (m, 1 H), 4.73-4.27 (m, 1 H), 3.83- 3.42 (m, 2H), 3.12-2.64 (m, 7H), 2.08-1.62 (m, 3H). LR ESI MS: m/z calcd for C30H32N6O4 [M+H]⁺, 553.62.

[00642] 6-(5-(3-(8-Hydroxy-4-oxo-3,4-dihydroquinazolin-2-yl)propanoyl)-

2,5-diazabicyclo[2.2.2]octan-2-yl)-A/-methylnicotinamide (320)

[00643] Prepared using General Procedure T, from compound 319. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.43 (s, 1 H), 7.90-7.73 (m, 1 H), 7.53-7.40 (m, 1 H), 7.23- 6.97 (m, 2H), 6.47-6.32 (m, 1 H), 4.93-4.63 (m, 2H), 3.76 (s, 1 H), 3.67-3.36 (m, 2H), 3.26-3.16 (m, 1 H), 3.04-2.84 (m, 2H), 2.78 (s, 4H), 2.06-1.65 (m, 4H), 1.35-1.07 (m, 3H), 0.86-0.69 (m, 1 H). LR ESI MS: m/z calcd for C24H26N6O4 [M+H]⁺, 463.50.

[00644] Preparation of 2-(3-(6-Hydroxy-4-oxo-3,4-dihydroquinazolin-2- yl)propanoyl)-1 ,2,3,4-tetrahydroisoquinoline-7-carbonitrile (325)

[00645] 2-Amino-5-(benzyloxy)benzamide (321)

[00646] Prepared using General Procedure E, from 2-Amino-5- hydroxybenzamide. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.69-7.62 (m, 1 H), 7.53-7.29 (m, 7H), 5.21 (s, 2H). LR ESI MS: m/z calcd for Ci4Hi4N₂O₂ [M+H]⁺, 243.27.

[00647] 4-(4-(Benzyloxy)-2-carbamoylphenylamino)-4-oxobutanoic acid

(322) [00648] Prepared using General Procedure U, from compound 321 and Succinic anhydride. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.23-8.15 (m, 1 H), 7.50-7.29 (m, 6H), 7.20-7.12 (m, 1 H), 5.13 (s, 2H), 2.74-2.65 (m, 4H). LR ESI MS: m/z calcd for C18H18N2O5 [M-H]; 341.34.

[00649] 3-(6-(Benzyloxy)-4-oxo-3,4-dihydroquinazolin-2-yl)propanoic acid (323)

[00650] Prepared using General Procedure S, from compound 322. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.22 (brs, 1 H), 7.60-7.28 (m, 8H), 5.21 (s, 2H), 2.90- 2.66 (m, 4H). LR ESI MS: m/z calcd for C18H16N2O4 [M-H]; 323.33.

[00651] 2-(3-(6-(Benzyloxy)-4-oxo-3,4-dihydroquinazolin-2-yl)propanoyl)- 1 ,2, 3, 4-tetrahydroisoquinoline-7 -carbonitrile (324)

[00652] Prepared using General Procedure N, from compound 323 and 1 , 2,3,4- Tetrahydroisoquinoline-7-carbonitrile. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.74- 7.58 (m, 2H), 7.56-7.44 (m, 3H), 7.44-7.27 (m, 6H), 5.18 (s, 2H), 4.81-4.59 (m, 2H), 3.80-3.62 (m, 2H), 3.05-2.69 (m, 7H). LR ESI MS: m/z calcd for C28H24N4O3 [M+H]⁺, 465.51.

[00653] 2-(3-(6-Hydroxy-4-oxo-3,4-dihydroquinazolin-2-yl)propanoyl)-

1 ,2, 3, 4-tetrahydroisoquinoline-7 -carbonitrile (325)

[00654] Prepared using General Procedure T, from compound 324. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.58-7.14 (m, 6H), 4.78-4.70 (m, 1 H), 3.92-3.77 (m, 2H), 3.12-2.96 (m, 6H), 2.95-2.85 (m, 2H). LR ESI MS: m/z calcd for C21H18N4O3 [M+H]⁺, 375.39.

[00655] Preparation of p-{4-[(E)-3-(8-Hydroxy-4-oxo-3H-quinazolin-2- yl)acryloyl]-1 -piperazinyl}benzonitrile (328)

[00656] 4-(2-Carbamoyl-6-hydroxyphenylamino)-4-oxo-2-butenoic acid

(326)

[00657] Synthesized using General Procedure U from 2-Amino-3- hydroxybenzamide and Maleic anhydride. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.27-7.20 (m, 1 H), 7.17-7.12 (m, 1 H), 7.08-7.03 (m, 1 H), 6.70 (d, 1 H), 6.35 (d, 1 H). LR ESI MS: m/z calcd for C11H9N2O5 [M-Hp, 249.1 found 249.7.

[00658] (E)-3-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)acrylic acid (327)

[00659] Synthesized using General Procedure S from compound 326. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 13.05 (br s, 1 H), 10.43 (s, 1 H), 7.57 (d, 1 H), 7.42 (t, 1 H), 7.28 (d, 1 H), 6.78 (d, 1 H), 6.44 (d, 1 H). LR ESI MS: m/z calcd for C11 H7N2O4 [M- Hp, 231.0 found 231.7.

[00660] p-{4-[(E)-3-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)acryloyl]-1- piperazinyl}benzonitrile (328)

[00661] Synthesized using General Procedure N from compound 327 and p-(1- Piperazinyl)benzonitrile. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 13.46 (br s, 1 H), 7.78 (dd, 1 H), 7.57-7.52 (m, 2H), 7.44-7.35 (m, 2H), 7.29-7.24 (m, 1 H), 6.90-6.85 (m, 2H), 6.77 (d, 1 H), 6.66 (d, 1 H). LR ESI MS: m/z calcd for C22H18N5O3 [M-Hp, 400.1 found 400.8; m/z calcd for C22H20N5O3 [M+H]⁺, 402.2 found 402.8.

[00662] Preparation of p-{4-[(E)-3-(8-Hydroxy-4-oxo-3H-quinazolin-2- yl)methacryloyl]-1 -piperazinyl}benzonitrile (331 )

[00663] (E)-4-(2-Carbamoyl-6-hydroxyphenylamino)-2-methyl-4-oxo-2- butenoic acid (329) [00664] Synthesized using General Procedure U from 2-Amino-3- hydroxybenzamide and 3-Methyl-2,5-furandione. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.73 (m, 1 H), 7.22 (d, 1 H), 7.15 (d, 1 H), 6.62 (s, 1 H), 2.15 (s, 3H).

[00665] (E)-3-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)methacrylic acid (330)

[00666] Synthesized using General Procedure S from compound 329. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.65 (br s, 1 H), 8.65 (s, 1 H), 7.54 (d, 1 H), 7.35 (m, 1 H), 7.25 (m, 1 H), 6.65 (s, 1 H), 2.11 (s, 3H).

[00667] p-{4-[(E)-3-(8-Hydroxy-4-oxo-3H-quinazolin-2-yl)methacryloyl]-1- piperazinyl}benzonitrile (331)

[00668] Synthesized using General Procedure N from compound 330 and p-(1- Piperazinyl)benzonitrile. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.65 (d, 1 H), 7.45 (d, 2H), 7.31 (m, 1 H), 7.20 (d, 1 H), 6.87 (d, 2H), 6.28 (s, 1 H), 3.88 (m, 2H), 3.54 (m, 2H), 3.44 (m, 2H), 3.22 (m, 2H). ES-MS: 414.8 [M-H]-.

[00669] Preparation of p-{4-[(E)-3-(8-Fluoro-4-oxo-3H-quinazolin-2- yl)acryloyl]-1 -piperazinyl}benzonitrile (333)

[00670] (E)-3-(8-Fluoro-4-oxo-3H-quinazolin-2-yl)acrylic acid (332)

[00671] Prepared using General Procedure U, from 2-Amino-3-fluorobenzamide and Maleic anhydride, followed by General Procedure S. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 8.08 (m, 1 H), 7.63 (m, 2H), 6.90 (d, 1 H), 6.56 (d, 1 H). ¹⁹F NMR (300 MHz, Methanol-cM): 5 (ppm) 125.3.

[00672] p-{4-[(E)-3-(8-Fluoro-4-oxo-3H-quinazolin-2-yl)acryloyl]-1- piperazinyl}benzonitrile (333)

[00673] Prepared using General Procedure N, from compound 332. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.94 (d, 1 H), 7.83 (d, 1 H), 7.63 (m, 1 H), 7.61 (d, 2H), 7.50 (m, 1 H), 7.22 (d, 1 H), 7.07 (d, 2H), 3.83 (br d, 4H), 3.38 (m, 4H). ¹⁹F NMR (300 MHz, DMSO-d6): 5 (ppm) 124.7. ES-MS: 402.8 [M-H]-.

[00674] Preparation of p-{4-[3-(8-Fluoro-4-thioxo-3H-quinazolin-2- yl)propionyl]-1 -piperazinyl}benzonitrile (337)

[00675] 2-Amino-3-fluorobenzenecarbothioamide (334)

[00676] 2-Amino-3-fluorobenzamide (1 equiv.) was dissolved in THF and Lawesson’s reagent (1 equiv.) was added as a solid at room temperature, the mixture was refluxed for 18h. Aqueous NaHCOa solution was added, and the product was extracted with EtOAc, dried over Na2SO4 and concentrated under vacuum to yield the title compound as a yellow solid, which was used in the next step without purification. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.20 (s, 1 H), 7.01 (m, 1 H), 6.50 (m, 1 H). ¹⁹F NMR (300 MHz, Methanol-cM): 5 (ppm) 136.1. ES-MS: 171.2 [M+H]+.

[00677] 4-(2-Carbamthioyl-6-fluorophenylamino)-4-oxobutyric acid (335)

[00678] Prepared using General Procedure U, from compound 334 and Succinic anhydride. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.52 (m, 1 H), 7.41 (m, 1 H), 7.22 (m, 1 H), 3.61 (m, 4H). ¹⁹F NMR (300 MHz, Methanol-cM): 5 (ppm) 137.3. ES-MS: 269.6 [M-H]’. [00679] 3-(8-Fluoro-4-thioxo-3H-quinazolin-2-yl)propionic acid (336)

[00680] Prepared using General Procedure S, from compound 335. ¹H NMR (300 MHz, Methanol-d4): 5 (ppm) 7.80 (d, 1 H), 7.30 (m, 1 H), 7.03 (m, 1 H), 3.51 (m, 2H), 3.2 (m, 2H). ¹⁹F NMR (300 MHz, Methanol-d4): 5 (ppm) 126.45. ES-MS: 251.7 [M-H]-.

[00681 ] p-{4-[3-(8-Fluoro-4-thioxo-3/7-quinazolin-2-yl)propionyl]-1 - piperazinyl}benzonitrile (337)

[00682] Prepared using General Procedure N, from compound 336. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 8.33 (d, 1 H), 7.63 (d, 4H), 7.52 (m, 1 H), 7.04 (d, 4H), 3.66 (br d, 4H), 3.44 (m, 4H). ¹⁹F NMR (300 MHz, DMSO-d6): 5 (ppm) 125.7. ES-MS: 420.28 [M-H]-.

[00683] Preparation of p-{4-[2,2,3,3-Tetrafluoro-3-(8-hydroxy-4-oxo-3H- quinazolin-2-yl)propionyl]-1 -piperazinyl}benzonitrile (341 )

[00684] 4-[2-(Benzyloxy)-6-carbamoylphenylamino]-2,2,3,3-tetrafluoro-4- oxobutyric acid (338)

[00685] Prepared using General Procedure U, from compound 145 and Tetrafluorosuccinic anhydride. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.39-7.22 (m, 6H), 7.16-7.10 (m, 1 H), 7.10-7.04 (m, 1 H), 5.10 (s, 2H). ¹⁹F NMR (300 MHz, Chloroform-d): 5 (ppm) -125.6, -125.7. LR ESI MS: m/z calcd for C18H13F4N2O5 [M-H]’ , 413.30 found 413.7. [00686] 3-[8-(Benzyloxy)-4-oxo-3H-quinazolin-2-yl]-2, 2,3,3- tetrafluoropropionic acid (339)

[00687] Prepared using General Procedure S, from compound 338. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.80 (d, 1 H), 7.48-7.19 (m, 7H), 5.27 (s, 2H). ¹⁹F NMR (300 MHz, Chloroform-d): 5 (ppm) -116.2, -119.1. LR ESI MS: m/z calcd for C18H11F4N2O4 [M-H]; 395.28 found 395.5.

[00688] p-(4-{3-[8-(Benzyloxy)-4-oxo-3H-quinazolin-2-yl]-2,2,3,3- tetrafluoropropionyl}-1 -piperazinyl)benzonitrile (340)

[00689] Prepared using General Procedure N, from compound 339 and p-(1- Piperazinyl)benzonitrile. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 7.88 (d, 1 H), 7.54- 7.45 (m, 4H), 7.45-7.37 (m, 2H), 7.32-7.20 (m, 2H), 6.89 (d, 2H), 6.78 (d, 2H), 5.21 (s, 2H), 3.89-3.81 (m, 2H), 3.69-3.60 (m, 2H), 3.51-3.43 (m, 4H). ¹⁹F NMR (300 MHz, Chloroform-d): 5 (ppm) -111.4, -116.2. LR ESI MS: m/z calcd for C29H22F4N5O3 [M-H]’ , 564.51 found 564.8.

[00690] p-{4-[2,2,3,3-Tetrafluoro-3-(8-hydroxy-4-oxo-3H-quinazolin-2- yl)propionyl]-1 -piperazinyl}benzonitrile (341 )

[00691] Prepared using General Procedure T, from compound 340. ¹H NMR (300 MHz, Methanol-cM): 5 (ppm) 7.71 (d, 1 H), 7.55-7.46 (m, 2H), 7.44-7.38 (m, 2H), 7.27 (d, 1 H), 6.89 (d, 2H), 3.96-3.87 (m, 2H), 3.78-3.71 (m, 2H), 3.48-3.33 (m, 4H). ¹⁹F NMR (300 MHz, DMSO-d6): 5 (ppm) -110.8, -114.4. LR ESI MS: m/z calcd for C22H16F4N5O3 [M-H]’, 474.39 found 474.3.

[00692] Preparation of p-(4-{2-[(8-Hydroxy-4-oxo-2- quinazolinyl)methoxy]acetyl}-1 -piperazinyl)benzonitrile (345)

[00693] {[A/-2-(Benzyloxy)-6-carbamoylphenyl carbamoyl]methoxy}acetic acid (342)

[00694] Prepared using General Procedure U, from compound 145 and 1 ,4- Dioxane-2, 6-dione. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.78 (br s, 1 H), 9.27 (br s, 1 H), 7.61 (m, 1 H), 7.50-7.06 (m, 8H), 5.15 (s, 2H), 4.17 (s, 2H), 4.15 (s, 2H). LR ESI MS: m/z calcd. for C18H18N2O6 [M+H]+ 357.8.

[00695] {[8-(Benzyloxy)-4-oxo-3,4-dihydro-2-quinazolinyl] methoxy}acetic acid (343)

[00696] Prepared using General Procedure S, from compound 342. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.87 (br s, 1 H), 12.23 (br s, 1 H), 7.76-7.60 (m, 7H), 7.59-7.24 (m, 1 H), 5.27 (s, 2H), 4.53 (s, 2H), 4.23 (s, 2H). LR ESI MS: m/z calcd. for C18H16N2O5 [M+H]+ 339.8.

[00697] p-[4-(2-{[8-(Benzyloxy)-4-oxo-3,4-dihydro-2- quinazolinyl]methoxy}acetyl)-1 -piperazinyl]benzonitrile (344)

[00698] Prepared using General Procedure N, from compound 343 and p-(1- Piperazinyl)benzonitrile. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.65-7.43 (m, 5H), 7.39-7.22 (m, 3H), 7.05-6.91 (m, 4H), 5.26 (s, 2H), 4.44 (s, 2H), 4.34 (s, 2H), 3.71 - 3.35 (m, 8H). LR ESI MS: m/z calcd. for C29H27N5O4 [M+H]+ 508.8.

[00699] p-(4-{2-[(8-Hydroxy-4-oxo-2-quinazolinyl)methoxy]acetyl}-1- piperazinyl)benzonitrile (345)

[00700] Prepared using General Procedure T, from compound 344. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.65-7.50 (m, 2H), 7.41 -7.25 (m, 1 H), 7.09-6.85 (m, 4H), 4.41 (m, 4H), 3.38 (m, 8H). LR ESI MS: m/z calcd. for C22H21N5O4 [M+H]+ 418.9.

[00701] Preparation of p-(4-{[3-(8-Hydroxy-4-oxo-2-quinazolinyl)-2,2- dimethylcyclopropyl]carbonyl}-1 -piperazinyl)benzonitrile (349)

[00702] 3-[A/-2-(Benzyloxy)-6-carbamoylphenylcarbamoyl]-2,2- dimethylcyclopropanecarboxylic acid (346)

[00703] Prepared using General Procedure U, from compound 145 and 6,6- Dimethyl-3-oxabicyclo[3.1.0]hexane-2, 4-dione. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 13.34 (br s, 1 H), 9.95 (br s, 1 H), 7.54-7.42 (m, 2H), 7.41 -7.14 (m, 5H), 7.13- 7.05 (m, 1 H), 5.14 (s, 2H), 2.14-2.01 (m, 1 H), 1.99-1.87 (m, 1 H), 1.21 (s, 3H), 1.18 (s, 3H). LR ESI MS: m/z calcd. for C21H22N2O5 [M+H]+ 381 .8.

[00704] 3-[8-(Benzyloxy)-4-oxo-3,4-dihydro-2-quinazolinyl]-2,2- dimethylcyclopropanecarboxylic acid (347)

[00705] Prepared using General Procedure S, from compound 346. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.71 (br s, 1 H), 8.07-6.88 (m, 8H), 5.29 (s, 2H), 2.43- 2.22 (m, 1 H), 2.20 (m, 1 H), 1.37 (s, 3H), 1.29 (s, 3H). LR ESI MS: m/z calcd. for C21H20N2O4 [M+H]+ 363.9.

[00706] p-[4-({3-[8-(Benzyloxy)-4-oxo-3,4-dihydro-2-quinazolinyl]-2,2- dimethylcyclopropyl}carbonyl)-1 -piperazinyl]benzonitrile (348)

[00707] Prepared using General Procedure N, from compound 347 and p-(1- Piperazinyl)benzonitrile. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.67-7.43 (m, 5H), 7.38-7.21 (m, 5H), 7.00-6.87 (m, 2H), 5.29-5.13 (m, 2H), 3.81-3.51 (m, 4H), 3.21 -2.97 (m, 4H), 2.31-2.16 (m, 2H), 1.33 (s, 3H), 1.30 (s, 3H). LR ESI MS: m/z calcd. for C32H31 N5O3 [M+H]+ 532.0.

[00708] p-(4-{[3-(8-Hydroxy-4-oxo-2-quinazolinyl)-2,2- dimethylcyclopropyl]carbonyl}-1 -piperazinyl)benzonitrile (349)

[00709] Prepared using General Procedure T, from compound 348. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.39-7.21 (m, 1 H), 7.12-6.77 (m, 4H), 6.78-6.65 (m, 2H), 3.11-2.79 (m, 4H), 2.78-2.65 (m, 1 H), 2.26-1.96 (m, 5H), 1.32 (s, 3H), 1.27 (s, 3H). LR ESI MS: m/z calcd. for C25H25N5O3 [M+H]+ 442.9.

[00710] Preparation of p-(4-{[2-(8-Hydroxy-4-oxo-2- quinazolinyl)cyclopropyl] carbonyl}-1 -piperazinyl)benzonitrile (353)

[00711] 2-[A/-2-(Benzyloxy)-6-carbamoylphenylcarbamoyl] cyclopropanecarboxylic acid (350)

[00712] Prepared using General Procedure U, from compound 145 and 3- Oxabicyclo[3.1.0]hexane-2, 4-dione. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.35 (br s, 1 H), 9.67 (br s, 1 H), 7.50-7.08 (m, 8H), 5.23 (s, 2H), 2.30-2.14 (m, 1 H), 2.07-1.93 (m, 1 H), 1.41-1.31 (m, 1 H), 1.27-1.15 (m, 1 H). LR ESI MS: m/z calcd. for C19H18N2O5 [M+H]+ 353.7.

[00713] 2-[8-(Benzyloxy)-4-oxo-3,4-dihydro-2-quinazolinyl] cyclopropanecarboxylic acid (351) [00714] Prepared using General Procedure S, from compound 350. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 12.36 (br s, 2H), 7.70-7.60 (m, 1 H), 7.56-7.46 (m, 2H), 7.44-7.27 (m, 5H), 5.28 (s, 2H), 2.62-2.52 (m, 1 H), 2.15-2.02 (m, 1 H), 1.83-1.72 (m, 1 H), 1 .44-1 .33 (m, 1 H). LR ESI MS: m/z calcd. for C19H16N2O4 [M+H]+ 335.8.

[00715] p-[4-({2-[8-(Benzyloxy)-4-oxo-3,4-dihydro-2- quinazolinyl]cyclopropyl} carbonyl)-1 -piperazinyl]benzonitrile (352)

[00716] Prepared using General Procedure N, from compound 351 and p-(1- Piperazinyl)benzonitrile. ¹H NMR (300 MHz, Chloroform-d): 5 (ppm) 10.49 (br s, 1 H), 7.83-7.76 (m, 1 H), 7.51-7.40 (m, 4H), 7.38-7.30 (m, 4H), 7.17-7.09 (m, 1 H), 6.81-6.71 (m, 2H), 5.30 (m, 2H), 4.00-3.63 (m, 4H), 3.51-3.08 (m, 4H), 2.85-2.72 (m, 1 H), 2.50- 2.38 (m, 1 H), 2.09-1.99 (m, 1 H), 1.71-1.64 (m, 1 H). LR ESI MS: m/z calcd. for C30H27N5O3 [M+H]+ 504.6.

[00717] p-(4-{[2-(8-Hydroxy-4-oxo-2-quinazolinyl)cyclopropyl] carbonyl}-1- piperazinyl)benzonitrile (353)

[00718] Prepared using General Procedure T, from compound 352. ¹H NMR (300 MHz, DMSO-d6): 5 (ppm) 7.60-7.49 (m, 2H), 7.47-7.37 (m, 1 H), 7.25-7.08 (m, 2H), 7.01-6.85 (m, 2H), 3.78-3.65 (m, 2H), 3.58-3.48 (m, 2H), 3.12-3.00 (m, 1 H), 2.90- 2.86 (m, 1 H), 1.97-1.84 (m, 1 H), 1.44-1.29 (m, 1 H) [Note: some piperazine protons were masked by DMSO signal], LR ESI MS: m/z calcd. for C23H21N5O3 [M+H]+ 414.9.

[00719] IN VITRO PARP-1 and PARP-2 INHIBITION ASSAYS

[00720] All the compounds herein were screened against PARP-1 and PARP-2 as well in order to determine the selectivity of inhibitors for PARP-1 over PARP-2.

[00721] PARP-1 Inhibitor Assay Design

[00722] Activities of compounds against PARP-1 were determined using the PARP-1 Colorimetric Assay Kit (Cat#80580) supplied by BPS Bioscience, USA. [00723] All samples and controls were performed in duplicates. The assay included a “Blank, a “Positive control”, and control inhibitor compound AZD5305 (CAS Number 2589531 -76-8). AZD5305 will inhibit the activity of PARP-1 at a wide range of concentrations from 1 nM to 10 pM. Serially dilute the stock AZD5305 or your PARP inhibitor(s) with 1X PARP Buffer and add to designated wells.

[00724] Step 1 : Coat histone solution using a 96-well transparent plate

[00725] 1) Dilute 5x histone mixture 1 :5 with PBS to make 1x histone mixture

[00726] 2) Add 50 pL of histone mixture to each well and incubate at 4°C overnight

[00727] 3) Wash the plate three times using 200 pL of PBST buffer (1x PBS containing 0.05% Tween 20) per well.

[00728] 4) Tap the plate onto clean paper towel to remove the liquid.

[00729] 5) Block the wells by adding 200 pL of Blocking buffer 3 to every well.

Incubate at room temperature for at least 90 minutes.

[00730] 6) Wash the plate three times with 200 pL/well of PBST buffer.

[00731] 7) Tap the plate onto clean paper towel to remove the liquid.

[00732] Step 2: Ribosylation reaction

[00733] 1) Prepare a fresh solution of 10 mM DTT in water.

[00734] 2) Dilute Activated DNA 1 :32 with PBS.

[00735] 3) Prepare the Master Mix (25 pL/well): N wells x (2.5 pL of 10x PARP buffer + 5 pL of PARP Substrate Mixture 1 + 5 pL of diluted Activated DNA + 10 pL of water + 2.5 pL of 10 mM fresh DTT).

[00736] 4) Add 25 pL of Master Mix to every well.

[00737] 5) Prepare 1x PARP buffer with DTT. Dilute 10x PARP assay buffer to

1x PARP assay buffer containing DTT by adding 1 volume of 10x PARP assay buffer + 1 volume of 10 mM DTT + 8 volumes of water.

[00738] 6) Add 5 pL of Test Inhibitor to each well labeled as “Test Inhibitor.”

[00739] For the “Positive Control" and “Blank,” add 5 pL of the same diluent solution used to dilute the inhibitor, but without inhibitor (Diluent Solution). [00740] 7) Thaw PARP-1 enzyme on ice. Briefly spin the tube containing the enzyme to recover the full content of the tube. Calculate the amount of PARP-1 required for the assay and dilute enzyme to 0.23 ng/pL with 1x PARP buffer with DTT. The final concentration of PARP-1 will be 1 nM. Aliquot the remaining undiluted PARP- 1 enzyme into aliquots and store at -80°C.

[00741] 8) Initiate the reaction by adding 20 pL of diluted PARP-1 enzyme to the wells designated “Positive Control” and "Test Inhibitor."

[00742] To the wells designated as "Blank," add 20 pL of 1x PARP buffer with DTT.

[00743] Incubate at room temperature for 1 hour.

[00744] 9) Wash the plate three times with 200 pL PBST buffer and tap the plate onto clean paper towel as described above.

[00745] Step 3: Detection

[00746] 1) Dilute Streptavidin-HRP 1 :50 in Blocking buffer 3.

[00747] 2) Add 50 pL of diluted Streptavidin-HRP to each well. Incubate for 30 minutes at room temperature.

[00748] 3) Wash three times with 200 pL PBST buffer and tap the plate onto clean paper towel.

[00749] 4) Add 100 pL of the colorimetric HRP substrate to each well and incubate the plate at room temperature until blue color is developed in the positive control well. For PARP-1 , it normally takes 15-20 min to fully develop the color. However, the optimal incubation time may vary and should be determined empirically by the user.

[00750] 5) After the blue color is developed, add 100 pL of 2 M sulfuric acid to each well. Read the absorbance at 450 nm using UVA/is spectrophotometer microplate reader. The negative control-blank well should be - 0.05 absorbance at 450 nm. Alternatively, the plate may be read at 650 nm without adding 2 M sulfuric acid, but the Signal-to-Background ratio will be decreased.

[00751] PARP-2 Inhibitor Assay Design [00752] Activities of compounds against PARP-2 were determined using the PARP-2 Colorimetric Assay Kit (Cat#80581) supplied by BPS Bioscience, USA.

[00753] All samples and controls were performed in duplicates. The assay included a “Blank, a “Positive control”, and control inhibitor compound AZD5305 (CAS Number 2589531 -76-8). AZD5305 will inhibit the activity of PARP-2 at a wide range of concentrations from 1 nM to 10 pM. Serially dilute the stock AZD5305 or your PARP inhibitor(s) with 1X PARP Buffer and add to designated wells.

[00754] Step 1 : Coat histone solution using a 96-well transparent plate

[00755] 1) Dilute 5x histone mixture 1 :5 with PBS to make 1x histone mixture

[00756] 2) Add 50 pL of histone mixture to each well and incubate at 4°C overnight

[00757] 3) Wash the plate three times using 200 pL of PBST buffer (1x PBS containing 0.05% Tween 20) per well.

[00758] 4) Tap the plate onto clean paper towel to remove the liquid.

[00759] 5) Block the wells by adding 200 pL of Blocking buffer 3 to every well.

Incubate at room temperature for at least 90 minutes.

[00760] 6) Wash the plate three times with 200 pL/well of PBST buffer.

[00761] 7) Tap the plate onto clean paper towel to remove the liquid.

[00762] Step 2: Ribosylation reaction

[00763] 1) Prepare a fresh solution of 10 mM DTT in water.

[00764] 2) Dilute Activated DNA 1 :32 with PBS.

[00765] 3) Prepare the Master Mix (25 pL/well): N wells x (2.5 pL of 10x PARP buffer + 5 pL of PARP Substrate Mixture 1 + 5 pL of diluted Activated DNA + 10 pL of water + 2.5 pL of 10 mM fresh DTT).

[00766] 4) Add 25 pL of Master Mix to every well.

[00767] 5) Prepare 1x PARP buffer with DTT. Dilute 10x PARP assay buffer to

1x PARP assay buffer containing DTT by adding 1 volume of 10x PARP assay buffer + 1 volume of 10 mM DTT + 8 volumes of water.

[00768] 6) Add 5 pL of Test Inhibitor to each well labeled as “Test Inhibitor.” [00769] For the “Positive Control" and “Blank,” add 5 pL of the same diluent solution used to dilute the inhibitor, but without inhibitor (Diluent Solution).

[00770] 7) Thaw PARP-2 enzyme on ice. Briefly spin the tube containing the enzyme to recover the full content of the tube. Calculate the amount of PARP-2 required for the assay and dilute enzyme to 0.23 ng/pL with 1x PARP buffer with DTT. The final concentration of PARP-2 will be 1 nM. Aliquot the remaining undiluted PARP- 2 enzyme into aliquots and store at -80°C.

[00771] 8) Initiate the reaction by adding 20 pL of diluted PARP-2 enzyme to the wells designated “Positive Control” and "Test Inhibitor."

[00772] To the wells designated as "Blank," add 20 pL of 1x PARP buffer with DTT.

[00773] Incubate at room temperature for 1 hour.

[00774] 9) Wash the plate three times with 200 pL PBST buffer and tap the plate onto clean paper towel as described above.

[00775] Step 3: Detection

[00776] 1 ) Dilute Streptavidin-HRP 1 :50 in Blocking buffer 3.

[00777] 2) Add 50 pL of diluted Streptavidin-HRP to each well. Incubate for 30 minutes at room temperature.

[00778] 3) Wash three times with 200 pL PBST buffer and tap the plate onto clean paper towel.

[00779] 4) Add 100 pL of the colorimetric HRP substrate to each well and incubate the plate at room temperature until blue color is developed in the positive control well. For PARP-2, it normally takes 15-20 min to fully develop the color. However, the optimal incubation time may vary and should be determined empirically by the user.

[00780] 5) After the blue color is developed, add 100 pL of 2 M sulfuric acid to each well. Read the absorbance at 450 nm using UVA/is spectrophotometer microplate reader. The negative control-blank well should be - 0.05 absorbance at 450 nm. Alternatively, the plate may be read at 650 nm without adding 2 M sulfuric acid, but the Signal-to-Background ratio will be decreased. [00781] PARP-1 and PARP-2 BIOLOGICAL RESULTS

[00782] The Table below provides data on the PARP-1 and PARP-2 IC50 results, the PARP-1 %-inhibition at different concentrations, and where available the selectivity for PARP-1 over PARP-2.

[00783] All publications, patents and patent applications cited above are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety.

[00784] Although preferred embodiments of the invention have been described herein in detail, it will be understood by those skilled in the art that variations may be made thereto without departing from the spirit of the invention or the scope of the appended claims.

Claims

WE CLAIM:

1. A compound of formula (I):

or a pharmaceutically acceptable salt or pro-drug thereof, wherein

R¹ and R² independently are H, halogen, a Ci-6-substituent optionally having one or more heteroatoms, OR¹¹, or NR¹²R¹³;

R¹¹, R¹² and R¹³ are independently selected from H, benzyl, a Ci-6-substituent optionally having one or more heteroatoms, C(=O)R¹⁴, C(=O)OR¹⁵, or C(=O)NR¹⁶R¹⁷;

R¹⁴, R¹⁵, R¹⁶, and R¹⁷ are independently selected from H, benzyl, an optionally substituted aromatic ring, an optionally substituted heteroaromatic ring, or a Ci-6-substituent optionally having one or more heteroatoms;

X is O, S, or NH;

L¹ is absent or is NH, and when L¹ is absent, the quinazolinone moiety is bonded to L²;

L² is a Ci-3-chain optionally substituted with one or more methyl or methoxy, and L² being further optionally substituted with one or more heteroatoms;

L³ is absent or is NH, O, S;

L⁴ is absent or is - CH2-; L⁵ is absent or is C(=O), C(=S), S(=O) or S(=O)2; wherein when L³ is absent, L² is bonded to L⁴; when L³ and L⁴ are absent, L² is bonded to L⁵; when L³, L⁴ and L⁵ are absent, L² is bonded to R; when L⁴ is absent, L³ is bonded to L⁵; when L⁴ and L⁵ are absent, L³ is bonded to R; and when L⁵ is absent, L⁴ is bonded to R; or

L¹, L², L³, L⁴ and L⁵ together form a three to six membered ring optionally having one or more heteroatoms; and

R is H or a Ci-20-substituent optionally having one or more heteroatoms.

2. The compound of formula (I) as defined in claim 1 , or a pharmaceutically acceptable salt or pro-drug thereof, wherein

R¹ and R² independently are H, halogen, or OR¹¹;

R¹¹ is H, benzyl, or a Ci-6-substituent optionally having one or more heteroatoms; and

X is O, or S.

3. The compound of formula (I) as defined in claim 1 or 2, or a pharmaceutically acceptable salt or pro-drug thereof, wherein the compound of formula (I) comprises:

represents the point of bonding to L¹, and if L¹ is absent,

represents the point of bonding to L².

4. The compound of formula (I) as defined in any one of claims 1 to 3, or a pharmaceutically acceptable salt or pro-drug thereof, wherein in the compound of formula (I), the quinazolinone moiety bonded to -L¹-L²-L³-L⁴-L⁵- is:

represents the point of bonding to R.

5. The compound of formula (I) as defined in any one of claims 1 to 4, or a pharmaceutically acceptable salt or pro-drug thereof, wherein R is:

6. The compound of formula (I) as defined in any one of claims 1 to 5, or a pharmaceutically acceptable salt or pro-drug thereof, wherein the structure formed by -L¹-L²-L³-L⁴-L⁵-R is:

7. The compound of formula (I) as defined in any one of claims 1 to 6, or a

5 pharmaceutically acceptable salt or pro-drug thereof, wherein the compound is:

£Z

8. A method for treating diseases mediated by PARP-1 protein comprising administering to a mammal in need thereof an effective amount of the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, as defined in any one of claims 1 to 7, or combination thereof.

9. The method according to claim 8, wherein the mammal in need is a human.

10. The method according to claim 8 or 9, wherein the disease mediated by PARP-

1 protein is cancer, cardiovascular diseases, nervous system injury or inflammation.

11. The method according to claim 10, wherein the cancer is a carcinomas, such as bladder, breast, colon, kidney, liver, lung, including small cell lung cancer, esophagus, gall-bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin, including squamous cell carcinoma; hematopoietic tumors of lymphoid lineage, including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell-lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkitt's lymphoma; hematopoietic tumors of myeloid lineage, including acute and chronic myelogenous leukemias, myelodysplastic syndrome and promyelocytic leukemia; tumors of mesenchymal origin, including fibrosarcoma, Ewing's sarcoma and rhabdomyosarcoma; tumors of the central and peripheral nervous system, including astrocytoma, neuroblastoma, glioma and schwannomas; and other tumors, including melanoma, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, keratoxanthoma, thyroid follicular cancer and Kaposi's sarcoma.

12. The method according to claim 8 or 9, wherein the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, as defined in any one of claims 1 to 7, or combination thereof, is administered in combination with radiation therapy or chemotherapy regimen for simultaneous, separate or sequential use in anticancer therapy.

13. An in vitro method for inhibiting PARP-1 protein activity, comprising contacting the said protein with an effective amount of the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, as defined in any one of claims 1 to 7, or combination thereof.

14. A pharmaceutical composition comprising a therapeutically effective amount of the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, as defined in any one of claims 1 to 7, or combination thereof, and at least one pharmaceutically acceptable excipient, carrier or diluent.

15. The pharmaceutical composition according to claim 14, further comprising one or more chemotherapeutic agents.

16. The pharmaceutical composition according to claim 15, wherein the chemotherapeutic agent is an alkylating agent.

17. The pharmaceutical composition according to claim 16, wherein the alkylating agent is temozolomide.

18. A product comprising the compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, as defined in any one of claims 1 to 7, or combination thereof, and one or more chemotherapeutic agents, as a combined preparation for simultaneous, separate or sequential use in anticancer therapy.

19. The product according to claim 18, wherein the chemotherapeutic agent is an alkylating agent.

20. The product according to claim 19, wherein the alkylating agent is temozolomide.

21. The compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, as defined in any one of claims to 1 to 7, or combination thereof for use as a medicament.

22. The compound of formula (I), pharmaceutically acceptable salt or pro-drug thereof, as defined in any one of claims to 1 to 7, or combination thereof, for use in treating cancer.

23. Use of the compound of formula (I), pharmaceutically acceptable salt or prodrug thereof, as defined in any one of claims to 1 to 7, or combination thereof, in the manufacture of a medicament for treatment of cancer.

24. Use of the compound of formula (I), pharmaceutically acceptable salt or prodrug thereof, as defined in any one of claims to 1 to 7, or combination thereof, for treating cancer.