US 20150353527A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0353527 A1 Zisman (43) Pub. Date: Dec. 10, 2015
(54) NON-SELECTIVE KINASE INHIBITORS Publication Classification (71) Applicant: arence ZIAMAN, Slingerlands, NY (51) Int. Cl. (US) C07D 40/12 (2006.01) (72) Inventor: Lawrence S Zisman, Slingerlands, NY C07D 24/20 (2006.01) (US) C07D 403/2 (2006.01) (73) Assignee: PULMOKINE, INC. Slingerlands, NY (52) U.S. Cl. (US) CPC ...... C07D401/12 (2013.01); C07D 403/12 (21) Appl. No.: 14f76O139 (2013.01); C07D 241/20 (2013.01) y x- - - 9 (22) PCT Filed: Jan. 9, 2014 (57) ABSTRACT (86). PCT No.: PCT/US1.4/10778 T.S. ). L. 9, 2015 Disclosed herein are compounds, compositions, and methods e 19 for preventing and treating proliferative diseases associated Related U.S. Application Data with aberrant receptor tyrosine kinase (RTK) activity. The (60) Provisional application No. 61/751.217, filed on Jan. therapeutic indications described herein more specifically 10, 2013, provisional application No. 61/889,887, relate to the non-selective inhibition of RTKs associated with filed on Oct. 11, 2013. vascular and pulmonary disorders. Patent Application Publication Dec. 10, 2015 Sheet 1 of 26 US 2015/0353527 A1
FIG. 1A
$: 8:3 $38: Coacentration (nM)
FG, B
C s C am
u was e D 9 9 A.
Concentration (nM) Patent Application Publication Dec. 10, 2015 Sheet 2 of 26 US 2015/0353527 A1
F.G. 1 C
Concentration (in M)
F.G. 1D
c C R sa
spagh d g e d A. :::::: Concentration (in M) Patent Application Publication Dec. 10, 2015 Sheet 3 of 26 US 2015/0353527 A1
FG. 2A 120% 5 IC50 (uM) || | 100% PK10453: 0.590 3094 || Imatinib: 0.434 a 60% g xxPK10453 N 40% f xximatinib O 20% SS O O% 5.00 O.50 O.05
F.G. 2B
120% IC50 (uM) 100% PK10453: 0.33O matinib: 0.620 80% 60% 40% 20%
5.00 O.50 O.05 Patent Application Publication Dec. 10, 2015 Sheet 4 of 26 US 2015/0353527 A1
IC50 (M) PK10453: O. 132
S. 100% natinib:Mia 1.81 t g 80% C 60%
S. 40% & PKO453 a ximatinib S. 20% - - -
SS -- O% &x-H 5.OOO 0.500 O.OSO 0.005 Dose (M)
F.G. 2) IC50 (uM) 120.0% PK10453 0.429 9 imatinib 3.25 it 100.0% ------2 80.0% alf 60.0% ». PK10453 40.0% SS 20.0% 0.0% 4 H Patent Application Publication Dec. 10, 2015 Sheet 5 of 26 US 2015/0353527 A1
FIG. 2E
PDGF 10 ng/ml + + + + + + + + Drug (uM) 67 34 17 84 A2 21 1 OS PK10453 pAKT(S473) PDGFAA matinib
PK10453
pAKTCT308) PDGFAA
PK10453 pAKT(S473) PDGF BB
PK10453 pAKT(T308) PDGF BB Patent Application Publication Dec. 10, 2015 Sheet 6 of 26 US 2015/0353527 A1
FG, 3
16OO
FIG. 4A ·Æ}.**~*~~~~~~~~);~:O,^~~~~); Olung plV 2OO
40 60 Time (min) Patent Application Publication Dec. 10, 2015 Sheet 7 of 26 US 2015/0353527 A1
F.G. 4B 3OOO 28OO . 2600 2400 2200 2000 1800 lung piNH Plasma piNH
1600 1400 1200 1000 800 600 400 200 r O .. : O 20 40 60 80 Time (min)
F.G. 5A 100 -
Patent Application Publication Dec. 10, 2015 Sheet 8 of 26 US 2015/0353527 A1
BEST AWAABE MAGE F.G. SB ? :
FG, SC
9/27 FIG.SD .2 Patent Application Publication Dec. 10, 2015 Sheet 9 of 26 US 2015/0353527 A1
BES AWAABEMAGE
FIG. 6A
60
4. O
Treatment Day
10/27 FIG. 6B Patent Application Publication Dec. 10, 2015 Sheet 10 of 26 US 2015/0353527 A1
BES AVAILABLE IMAGE
320
O O
8 Patent Application Publication Dec. 10, 2015 Sheet 11 of 26 US 2015/0353527 A1
FG
F.G. 7C
O2
O
D4 D8 Patent Application Publication Dec. 10, 2015 Sheet 12 of 26 US 2015/0353527 A1
BEST AWAABE MAGE
g
FIG. 8B
F.G. 8C F.G. 8D
Patent Application Publication Dec. 10, 2015 Sheet 13 of 26 US 2015/0353527 A1
BES AWAABLE IMAGE
F.G. 10A FG, 10B
14/27
FG. A F.G. B FG, 1C
Patent Application Publication Dec. 10, 2015 Sheet 14 of 26 US 2015/0353527 A1
BES AVALABEWAGE
F.G. 1D FIG. 11E F.G. 1 F
F.G. 1G F.G. 11H F.G. 1
15/27
F.G. 12A F.G. 12B Patent Application Publication Dec. 10, 2015 Sheet 15 of 26 US 2015/0353527 A1
BEST AWAABE MAGE
FIG. 12C
F.G. 12E FG, 12F
F.G. 13B Patent Application Publication Dec. 10, 2015 Sheet 16 of 26 US 2015/0353527 A1
BEST AWAABE MAGE
F.G. 13C F.G. 13D
F.G. 14A
2,000: pAKThr3O8) 1,500 : ta. AK
1,000 500
A. 4.5 5 5.5 6 6.5 7 7.5
17/27 F.G. 4B pAKT(Thr308) 3,000 ota AK Patent Application Publication Dec. 10, 2015 Sheet 17 of 26 US 2015/0353527 A1
F.G. 14D
500 pAKT (Ser473
400 a AK
300
200
100
Patent Application Publication Dec. 10, 2015 Sheet 18 of 26 US 2015/0353527 A1
F.G. 4F
V D4 D8
FIG. SA o STAT3 pSTAT3 3,000
2000:
1,000 Patent Application Publication Dec. 10, 2015 Sheet 19 of 26 US 2015/0353527 A1
FIG. 15B 4,000
3,000
2000
1,000
8 0.4 8 8:
O. 3
-- --
Patent Application Publication Dec. 10, 2015 Sheet 20 of 26 US 2015/0353527 A1
12,000 FIG. 16A ppERK1
800 800 400 200
1. i FIG. 16C Patent Application Publication Dec. 10, 2015 Sheet 21 of 26 US 2015/0353527 A1
16 OOC F.G. 6) ERK1 14DDC 12DDC OOOC
6
0,900 0.800
0,700 w O,600 0,500 0.400 O3OO ---- 0.200 0.00 0,000 : V
Patent Application Publication Dec. 10, 2015 Sheet 22 of 26 US 2015/0353527 A1
F.G. 7A PDGFAA 3.3 ...... O3: X3:33 s: : Kis
FG. 17B PDGFBB
* "...... - M M M M M M M M M M M M M M M M M M M M M M M M M M M M ::iii:
PDGFAA iratin: K{2:33. K. Patent Application Publication Dec. 10, 2015 Sheet 23 of 26 US 2015/0353527 A1
FIG. 17D PDGFBB
...... in ...... K.3
33
3.3 : SF OLM :: ; ; ; 8x88; ; ;8:38: 1 ATN 8 3. 888 fiv. 388 88 ...... F.G. 18A
: ; ; ; ; ;- .. ::: ;
Drug Concentration (micromolar) *:::{3487 33 ri::::::gr:-i: ...atio:-- 3:gf::::... $33: 33 iciatic: ; 3 :::::t&s w8x 8K 2.337 33 ririt: gir:-ir:::::::::cr+ 33rgfrii. 8.338 88 ric: 3:3: ... :::::::::s -- 3:a::::: 383 ri: pre-i:33:iors. 33rig fr; . .338 38 ric:3:33:3: ... ::::::t&s Patent Application Publication Dec. 10, 2015 Sheet 24 of 26 US 2015/0353527 A1
F W P K ... O A. S. 3 C R P K . . . . . F.G. 18B + 1 O N G / M L PD G F B B s
.3 k A .5
. . . O. O. Drug Concentration (micromolar)
9 se (A) PK1045330 minute pre-incubation + 1Ong/mL PDGF BB
use (B) PK1045330 minute pre-incubation + 10ng/mL PDGF BB arras PK1057130 minute pre-incubation + 1Ong/mL PDGF BB Patent Application Publication Dec. 10, 2015 Sheet 25 of 26 US 2015/0353527 A1
FG, 18C
; : : Drug Concentration (micromolar) i-K:3488 38 :::::::is re-ic: ;83tion - K3333333 finista 8-incubati 33;g fir: 383- 38 inciatio: g;; 3:3: 3:::::::::
F.G. 18) O o
Y. O o, N scoO C O SN Structure 2 - N Structure 2a
esN? O Structure 3 N ?SS1. Joro Structure 41.N 2 N
Patent Application Publication Dec. 10, 2015 Sheet 26 of 26 US 2015/0353527 A1
250 forgopogrom 3 : 3 & 3 & 8 & 33 33 3: 3 3: 3 F.G. 20 Treatment Day
& & 83-** x:x:::::::::::3
Treatment Day US 2015/0353527 A1 Dec. 10, 2015
NON-SELECTIVE KNASE INHIBITORS 0007. In concert with the development and administration of first generation RTK inhibitors, e.g., imatinib, RTKs have CROSS-REFERENCE TO RELATED evolved inhibitor resistance by acquiring certain mutations. APPLICATIONS See, e.g., Shah et al., Science, 305, 395-402 (2004). For 0001. This application is a 35 U.S.C. 371 National Stage example, in diseased patients refractory to certain kinase entry of PCT/US2014/010778, filed Jan. 9, 2014, which inhibitors, e.g., imatinib, it has been shown that the hydro claims priority to U.S. Provisional Application No. 61/751, phobic pocket 'gatekeeper residue' frequently possesses 217 filed Jan. 10, 2013, and U.S. Provisional Application No. mutations. See Pao et al., PLos Med. 2(3):e73 (2005). Such 61/889,887, filed Oct. 11, 2013, the entire contents of which mutations have been identified with respect to ABL, i.e., at the are hereby incorporated by reference in their entirety. T315 residue, and at analogous positions in KIT, PDGFRO. EGFR, and other kinases. Id. Hence, new RTK inhibitors with STATEMENT OF GOVERNMENTSPONSORED Superior efficacy developed in model systems that phenotypi RESEARCH cally resemble human disease pathologyare required for pre venting and treating diseases possessing aberrant RTKactiv 0002 This invention was made with United States govern 1ty. ment support under Grant Number 1R43HL 102946-01 and 2R44HL 102946-02 awarded by the National Institute of SUMMARY OF THE INVENTION Health. The United States government has certain rights in the invention. 0008. In one aspect, the present disclosure provides a method of non-selective kinase receptor inhibition for treat FIELD OF THE INVENTION ing pulmonary disorders in a Subject, including: administer ing to the Subject a therapeutically effective amount of a 0003. The present disclosure relates generally to the treat compound of Structure 1, a tautomer, enantiomer, isomer or ment and prevention of disease associated with protein kinase Stereoisomer of the compound, a pharmaceutically accept activity. In particular, the present technology relates to thera able salt of the compound, tautomer, enantiomer, isomer or peutic indications of protein kinase inhibitors and methods Stereoisomer of the commound, or any mixtures thereof, for the treatment or prevention of pulmonary and vascular where Structure 1 has the formula: conditions, cancer, and other disorders.
BACKGROUND OF THE INVENTION Structure 1 0004. The following discussion of the background is RI merely provided to aid the reader in understanding the inven R2 tion and does not necessarily describe or constitute prior art. 21 R4 0005 Receptor tyrosine kinases (RTKs) are transmem brane polypeptides that regulate the regeneration, remodel 1S sk1 - ing, development and differentiation of cells and tissues. See, e.g., Mustonen et al., J. Cell Biology 129, 895-898 (1995); van der Geer et al. Ann Rev. Cell Biol. 10, 251-337 (1994). In R71 X nx2 X R5 addition to activating RTKs, polypeptide ligand growth fac tors and cytokines are capable of inducing conformation k changes in RTK external domains which results in receptor dimerization. Lymboussaki, Dissertation, Univ. of Helsinki, 0009 And where X is independently selected from C, N, Mol./Cancer Bio Lab and Dept. of Pathology, Haartman O, S or –CN: Institute (1999); Ullrich et al., Cell 61, 203-212 (1990). Cog 0010) R', R, and R mayy be the same or different and are nate RTK receptor-ligand binding, moreover, imparts recep independently selected from the group consisting of H, C, N, tor trans-phosphorylation at specific tyrosine residues and O, S, Cl, Br, F, I, CN, NO, -OH, -CH, -CF, Subsequent activation of the kinase catalytic domains, —C N C - groups, —C N—C(=O)— groups, thereby enabling Substrate phosphorylation and activation of —C(=O)R groups, N C(=O)R groups, C N C associated signaling cascades. Id. (=O)R groups, substituted and unsubstituted R groups, 0006 Aberrant RTKactivity, however, is associated with a substituted and unsubstituted R groups substituted with one variety of disease conditions and systemic delivery of certain or more of R. R', and R', substituted and unsubstituted RTK inhibitors have shown efficacy for specific disease con amidinyl groups, Substituted and unsubstituted guanidinyl ditions. In vivo assays to this end, including the murine mono groups, Substituted and unsubstituted primary, secondary, crotaline (MCT) model system, have been employed for and tertiary alkyl groups, Substituted and unsubstituted aryl ascertaining whether putative RTK inhibitors would function groups, Substituted and unsubstituted alkenyl groups, Substi as therapeutic agents. Concerning preclinical drug candidate tuted and unsubstituted alkynyl groups, Substituted and efficacy, however, the MCT model has been criticized inas unsubstituted heterocyclyl groups, Substituted and unsubsti much as such a system fails to Substantiate certain human tuted aminoalkyl groups, Substituted and unsubstituted alky disease phenotypes, e.g., the development of neointimal and/ laminoalkyl groups, Substituted and unsubstituted dialky or plexiform lesions that are symptomatically comorbid with laminoalkyl groups, Substituted and unsubstituted Such diseases. Hence, this model is an imperfect system, arylaminoalkyl groups, Substituted and unsubstituted diary which may confound the etiological and pathological indica laminoalkyl groups, Substituted and unsubstituted (alkyl) tions of human disease. Thus, new or complementary model (aryl)aminoalkyl groups, Substituted and unsubstituted het systems are necessary for accurate and efficient drug devel erocyclylalkyl groups, Substituted and unsubstituted cyano opment. groups, Substituted and unsubstituted pyrimidinyl groups, US 2015/0353527 A1 Dec. 10, 2015
Substituted and unsubstituted cyano (aryl) groups, Substituted alkyl groups, —N(aryl)O-alkyl groups, —N(alkyl)O-aryl and unsubstituted cyano (heterocyclyl) groups, and Substi groups, and —N(aryl)O-aryl groups; tuted and unsubstituted cyano-pyrimidinyl groups; I0012 R is selected from the group consisting of R. R. 0011 R,R,R, and R7, may be the same or different and R,R,R,R,R, H, absent, -C=C, substituted and unsub are independently selected from the group consisting of H, stituted heterocyclyl groups, Substituted and unsubstituted Cl, Br, F, I, —CN, NO. —OH, -CH, —CF, NH, aryl groups, substituted and unsubstituted heterocyclyl(R) —C=N, -C=N groups, —C N C - groups, groups, substituted and unsubstituted heterocyclyl(R') —C N C(=O)— groups, —C N C(=O) C F. groups, substituted and unsubstituted heterocyclyl(R') —C N C(=O)—C—C, substituted and unsubstituted groups, substituted and unsubstituted heterocyclyl(R)(R') alkyl groups, Substituted and unsubstituted aryl groups, Sub groups, substituted and unsubstituted heterocyclyl(R)(R') stituted and unsubstituted heterocyclyl groups, alkoxy groups, substituted and unsubstituted heterocyclyl(R')(R') groups, aryloxy groups, Substituted and unsubstituted hetero groups, substituted and unsubstituted heterocyclyl(R9)(R') cyclylalkyl groups, Substituted and unsubstituted aminoalkyl (R') groups, substituted and unsubstituted —C(=O)-het groups, Substituted and unsubstituted alkylaminoalkyl erocyclyl(R) groups, substituted and unsubstituted groups, Substituted and unsubstituted dialkylaminoalkyl —C(=O)-heterocyclyl(R') groups, substituted and unsub groups, Substituted and unsubstituted arylaminoalkyl groups, stituted—C(=O)-heterocyclyl(R') groups, substituted and Substituted and unsubstituted diarylaminoalkyl groups, Sub unsubstituted —C(=O)-heterocyclyl(R)(R') groups, sub stituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, stituted and unsubstituted—C(=O)-heterocyclyl (R)(R') Substituted and unsubstituted alkylamino groups, Substituted groups, Substituted and unsubstituted —(=O)-heterocyclyl and unsubstituted arylamino groups, Substituted and unsub (R') groups, substituted and unsubstituted —(=O)-hetero stituted dialkylamino groups, Substituted and unsubstituted cyclyl(R)(R')(R') groups, substituted and unsubstituted diarylamino groups, Substituted and unsubstituted (alkyl) aryl(R) groups, substituted and unsubstituted aryl(R') (aryl)amino groups, —C(=O)H, —C(=O)-alkyl groups, groups, substituted and unsubstituted aryl (R') groups, sub —C(=O)-aryl groups, —C(=O)C)-alkyl groups, —C(=O) stituted and unsubstituted aryl (R)(R') groups, substituted O-aryl groups, —C(=O)NH2. —C(=O)NH(alkyl) groups, and unsubstituted aryl (R)(R') groups, substituted and —C(=O)NH(aryl) groups, —C(=O)N(alkyl) groups, unsubstituted aryl(R')(R') groups, substituted and unsub C(=O)-aryl groups, —C(=O)NH2. —C(=O)NH(alkyl) stituted aryl (R)(“")(R') groups, substituted and unsubsti groups, —C(=O)NH(aryl) groups, —C(=O)N(alkyl). tuted—(=O)-aryl(R) groups, substituted and unsubstituted groups, —CGEO)N(aryl) groups, —CGEO)N(alkyl)(aryl) —(=O)-aryl(R') groups, substituted and unsubstituted groups, —C(=O)C)-alkyl groups, —C(=O)C-aryl groups, —(=O)-aryl(R') groups, substituted and unsubstituted —C(=O)-heterocyclyl groups, —C(=O)-O-heterocyclyl —(=O)-aryl(R)(R') groups, substituted and unsubstituted groups, —C(=O)NH(heterocyclyl) groups, —C(=O)-N —(=O)-aryl(R)(R') groups, substituted and unsubstituted (heterocyclyl) groups, C(=O) N(alkyl)(heterocyclyl) —C(=O)-aryl(R')(R') groups, and substituted/unsubsti groups, —C(=O)—N(aryl)(heterocyclyl) groups, Substi tuted —(=O)-aryl(R)(')(R') groups: tuted and unsubstituted heterocyclylaminoalkyl groups, Sub 0013 R. R', and R' may be the same or different and stituted and unsubstituted hydroxyalkyl groups, Substituted are independently selected from the group consisting of and unsubstituted alkoxyalkyl groups, Substituted and unsub absent, H, Cl, Br, F, I, —CN, NO. —OH, —CH, —CF, stituted aryloxyalkyl groups, and Substituted and unsubsti NH, —C(=O) , —C N R', -C=N-C groups, tuted heterocyclyloxyalkyl groups, Substituted and unsubsti —C N C(=O)— groups, —C N C(=O) C F. tuted diheterocyclylaminoalkyl, substituted and —C N C(=O)—C—C. —C—N groups, substituted and unsubstituted (heterocyclyl)(alkyl)aminoalkyl, substituted unsubstituted alkyl groups, Substituted and unsubstituted aryl and unsubstituted (heterocyclyl) (aryl)aminoalkyl, Substi groups, Substituted and unsubstituted heterocyclyl groups, tuted and unsubstituted alkoxyalkyl groups, Substituted and alkoxy groups, aryloxy groups, Substituted and unsubstituted unsubstituted hydroxyalkyl groups, Substituted and unsubsti heterocyclylalkyl groups, Substituted and unsubstituted ami tuted aryloxyalkyl groups, and Substituted and unsubstituted noalkyl groups, Substituted and unsubstituted alkylami heterocyclyloxyalkyl groups; -(alkyl)(aryl)aminoalkyl noalkyl groups, Substituted and unsubstituted dialkylami groups, —(=O)-heterocyclyl groups, —(=O)—O-hetero noalkyl groups, Substituted and unsubstituted arylaminoalkyl cyclyl groups, —C(=O)NH(heterocyclyl) groups, groups, Substituted and unsubstituted diarylaminoalkyl —C(=O)—N (heterocyclyl) groups, —C(=O)—N(alkyl) groups, Substituted and unsubstituted (alkyl)(aryl)ami (heterocyclyl) groups, —C(=O)—N(aryl)(heterocyclyl) noalkyl groups, Substituted and unsubstituted alkylamino groups, Substituted and unsubstituted heterocyclylami groups, Substituted and unsubstituted arylamino groups, and noalkyl groups, Substituted and unsubstituted hydroxyalkyl Substituted and unsubstituted dialkylamino groups, Substi groups, Substituted and unsubstituted alkoxyalkyl groups, tuted and unsubstituted aminoalkyl groups, Substituted and Substituted and unsubstituted aryloxyalkyl groups, and Sub unsubstituted (alkyl)(aryl)aminoalkyl groups, Substituted stituted and unsubstituted heterocyclyloxyalkyl groups, and unsubstituted alkylamino groups, Substituted and unsub —NH(alkyl) groups, —NH(aryl) groups, -N (alkyl) stituted arylamino groups, Substituted and unsubstituted groups, —N(aryl) groups, —N(alkyl)(aryl) groups, —NH dialkylamino groups, Substituted and unsubstituted diary (heterocyclyl) groups, —N(heterocyclyl)(alkyl) groups, lamino groups, Substituted and unsubstituted (alkyl)(aryl) —N(heterocyclyl)(aryl) groups, —N(heterocyclyl) groups, amino groups, —C(=O)H, —C(=O)-alkyl groups, Substituted and unsubstituted alkyl groups, Substituted and —C(=O)-aryl groups, —C(=O)C)-alkyl groups, —C(=O) unsubstituted aryl groups, Substituted and unsubstituted O-aryl groups, C(=O)NH2. —C(=O)NH(alkyl) groups, alkoxy groups, Substituted and unsubstituted aryloxy groups, —C(=O)NH(aryl) groups, —C(=O)N(alkyl) groups, substituted and unsubstituted heterocyclyl groups, NHOH, —C(=O)-aryl groups, —C(=O)NH2. —C(=O)NH(alkyl) —N(alkyl)OH groups, N(aryl)OH groups, —N(alkyl)O- groups, —C(=O)NH(aryl) groups, —C(=O)N(alkyl). US 2015/0353527 A1 Dec. 10, 2015 groups, —C(=O)N(aryl) groups, —C(=O)N(alkyl)(aryl) Substituted and unsubstituted pyrimidinyl groups, Substituted groups, —C(=O)C)-alkyl groups, —C(=O)C-aryl groups, and unsubstituted cyano (aryl) groups, Substituted and unsub —C(=O)-heterocyclyl groups, —C(=O) O-heterocyclyl stituted cyano (heterocyclyl) groups, and Substituted and groups, —C(=O)NH(heterocyclyl) groups, —C(=O)-N unsubstituted cyano-pyrimidinyl groups; (heterocyclyl) groups, —C(=O)—N(alkyl)(heterocyclyl) 10016 Q is selected from the groups consisting of absent, groups, —C(=O)—N (aryl)(heterocyclyl) groups, Substi H. Q', Q' (Q), -OH, alkoxy groups, aryloxy groups; and tuted and unsubstituted heterocyclylaminoalkyl groups, Sub I0017 Q is selected from the group consisting of absent, a stituted and unsubstituted cyano groups, Substituted and direct bond, H, C, Cl, Br, F. I. —CN, NO. —CH, —CF, unsubstituted pyrimidinyl groups, Substituted and unsubsti tuted cyano(aryl) groups, Substituted and unsubstituted cyano (heterocyclyl) groups, and Substituted and unsubstituted cyano-pyrimidinyl groups; I0014) R' is selected from the group consisting of absent, CF, C=N, C. N. C groups, —C N—C H, Cl, Br, F.I. —CN, NO. —OH, -CH, -CF, NH, (=O)— groups, —C N—C(=O) C F. —C N C (=O) , —C N R', -C=N. —C N C groups, (=O)—C=C. —OH, alkoxy groups, alkoxy groups, aryloxy —C N C(=O)— groups, —C N C(=O) C F. groups, methoxy groups, dimethoxy groups, methoxy phe —C N C(=O)—C—C. —C—N groups, —(=O)— nol, methoxy phenol groups, dimethoxy phenol, dimethoxy groups, —(=O)—C-groups, —(=O)—C=C. —S(=O) phenol groups, dimethoxy benzene, dimethoxy benzene — groups, S(=O). C-groups, —S(=O). C=C groups, Substituted and unsubstituted alkyl groups, Substi groups, —S(=O) -C=C-CH, alkoxy groups, aryloxy tuted and unsubstituted aryl groups, and Substituted and groups, Substituted and unsubstituted amidinyl groups, Sub unsubstituted heterocyclyl groups. The contents of the fore stituted and unsubstituted guanidinyl groups, Substituted and going paragraph (i.e., 0009) are hereinafter referred to as unsubstituted primary, secondary, and tertiary alkyl groups, “QXR". Substituted and unsubstituted aryl groups, Substituted and I0018. In illustrative embodiments, the structure of R has unsubstituted alkenyl groups, Substituted and unsubstituted the following formula: alkynyl groups, Substituted and unsubstituted heterocyclyl groups, Substituted and unsubstituted aminoalkyl groups, Substituted and unsubstituted alkylaminoalkyl groups, Sub R9 stituted and unsubstituted dialkylaminoalkyl groups, substi RQ CO tuted and unsubstituted arylaminoalkyl groups, Substituted Nx 2 and unsubstituted diarylaminoalkyl groups, Substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, Substituted and unsubstituted heterocyclylalkyl groups, Substituted and ls, unsubstituted cyano groups, Substituted and unsubstituted pyrimidinyl groups, Substituted and unsubstituted cyano (0019 where X is independently selected from C, N, O, S, (aryl) groups, Substituted and unsubstituted cyano (heterocy and –CN; R. R', and R'' may be the same or different and clyl) groups, and Substituted and unsubstituted cyano-pyrim are independently selected from the group consisting of H. C. idinyl groups; N, O, S, Cl, Br, F, I, CN, NO, OH, -CH, -CF, 0015 Q' is selected from the group consisting of a direct NH, -C(=O) , —C N R', -C=N, C-N C bond, H., Cl, Br, F, I, —CN, NO. —CH, —CF, NH, (=O) C F. —C N C(=O)—C—C, substituted and (=O) , —C N R', -C=N, —C N C groups, unsubstituted alkyl groups, Substituted and unsubstituted aryl groups, Substituted and unsubstituted heterocyclyl groups, —OH, alkoxy groups, aryloxy groups, Substituted and unsub stituted heterocyclylalkyl groups, Substituted and unsubsti tuted aminoalkyl groups, Substituted and unsubstituted alky laminoalkyl groups, Substituted and unsubstituted C=C. —OH, alkoxy groups, aryloxy groups, Substituted and dialkylaminoalkyl groups, Substituted and unsubstituted ary unsubstituted alkyl groups, Substituted and unsubstituted aryl laminoalkyl groups, Substituted and unsubstituted diarylami groups, Substituted and unsubstituted heterocyclyl groups, noalkyl groups, Substituted and unsubstituted (alkyl)(aryl) alkoxy groups, aryloxy groups, methoxy groups, dimethoxy aminoalkyl groups, Substituted and unsubstituted alkylamino groups, methoxyphenol, methoxyphenol groups, dimethoxy groups, Substituted and unsubstituted arylamino groups, and phenol, dimethoxy phenol groups, dimethoxy benzene, Substituted and unsubstituted dialkylamino groups, Substi dimethoxybenzene groups, methoxymethylbenzyl groups, tuted and unsubstituted cyano groups, Substituted and unsub Substituted and unsubstituted aralkyl groups, —NH2. Substi stituted pyrimidinyl groups, Substituted and unsubstituted tuted and unsubstituted heterocyclylalkyl groups, Substi cyano (aryl) groups, Substituted and unsubstituted cyano (het tuted/unsubstituted aminoalkyl groups, Substituted and erocyclyl) groups, and Substituted and unsubstituted cyano unsubstituted alkylaminoalkyl groups, Substituted and pyrimidinyl groups; and unsubstituted dialkylaminoalkyl groups, Substituted and I0020) R' is selected from the group consisting of unsubstituted arylaminoalkyl groups, Substituted and unsub —(=O)— groups, —(=O)—C-groups, —(=O)—C=C. stituted diarylaminoalkyl groups, Substituted and unsubsti —S(=O) groups, S(=O), C - groups, —S(=O) tuted (alkyl)(aryl)aminoalkyl groups, Substituted and unsub C=C- groups, —S(=O). C=C CH, -OH, stituted alkylamino groups, Substituted and unsubstituted alkoxy groups, aryloxy groups, Substituted and unsubstituted arylamino groups, and Substituted and unsubstituted dialky amidinyl groups, Substituted and unsubstituted guanidinyl lamino groups, Substituted and unsubstituted cyano groups, groups, Substituted and unsubstituted primary, secondary, US 2015/0353527 A1 Dec. 10, 2015
and tertiary alkyl groups, Substituted and unsubstituted aryl -continued groups, Substituted and unsubstituted alkenyl groups, Substi tuted and unsubstituted alkynyl groups, Substituted and unsubstituted heterocyclyl groups, Substituted and unsubsti o1 tuted aminoalkyl groups, Substituted and unsubstituted alky s N, and laminoalkyl groups, Substituted and unsubstituted dialky OH 1 laminoalkyl groups, Substituted and unsubstituted 2 arylaminoalkyl groups, Substituted and unsubstituted diary laminoalkyl groups, Substituted and unsubstituted (alkyl) O -C N (aryl)aminoalkyl groups, Substituted and unsubstituted het erocyclylalkyl groups, Substituted and unsubstituted cyano groups, Substituted and unsubstituted pyrimidinyl groups, OH Substituted and unsubstituted cyano (aryl) groups, Substituted and unsubstituted cyano (heterocyclyl) groups, and Substi tuted and unsubstituted cyano-pyrimidinyl groups. The con tents of the foregoing paragraph (i.e., 0010) are hereinafter Group B structures referred to as “QXR2. 0021. In illustrative embodiments, the structure of R is selected from Group A as shown. 0023. In some embodiments, the compound of Structure 1 is a compound of Structure 2, 2a, 3, 4 or 5, as shown below in Group C. Group A structures
Structure 2
OH
H N N HN n
2 21 O N Sa
Structure 2a o1 o1 H N N I0022. In illustrative embodiments, the structure of Q" or Q is selected from Group B as shown below, —CH, -OH, O CH, C N C(=O)—C=C, and - C N C 21 O % (=O) C F. Sa N
o1 o1 Structure 3 s o1 OH H N. N. o1 OH HN C Q?, Q?, US 2015/0353527 A1 Dec. 10, 2015
-continued 0026. In illustrative embodiments, the PAH selected from primary PAH, idiopathic PAH, heritable PAH, refractory Structure 4 PAH, BMPR2, ALK1, endoglin associated with hereditary hemorrhagic telangiectasia, endoglin not associated with O1 hereditary hemorrhagic telangiectasia, drug-induced PAH, OH and toxin-induced PAH, PAH associated with systemic scle H rosis, mixed connective tissue disease, HIV, hepatitis, and/or N N HN n portal hypertension. a O N 0027. In illustrative embodiments, the PAH is secondary H to pulmonary hypertension, congenital heart disease, a S N A w. hypoxia, chronic hemolytic anemia, newborn persistent pull O monary hypertension, pulmonary veno-occlusive disease (PVOD), pulmonary capillary hemangiomatosis (PCH), left Structure 5 heart disease pulmonary hypertension, Systolic dysfunction, o1 diastolic dysfunction, Valvular disease, lung disease, intersti tial lung disease, pulmonary fibrosis, Schistosomiasis, chronic obstructive pulmonary disease (COPD), sleep-disor H dered breathing, alveolar hypoVentilation disorders, chronic N N HN n exposure to high altitude, developmental abnormalities, chronic thromboembolic pulmonary hypertension (CTEPH), 2 21 O N pulmonary hypertension with unclear multifactorial mecha nisms, hematologic disorders, myeloproliferative disorders, Sa splenectomy, systemic disorders, sarcoidosis, pulmonary N Langerhans cell histiocytosis, lymphangioleimoyomatosis, Group C structures neurofibromatosis, vasculitis, metabolic disorders, glycogen storage disease, Gaucher disease, thyroid disorders, tumoral 0024. In illustrative embodiments, the compound of Struc obstruction, fibrosing mediastinitis, and/or chronic renal fail ture 1, 2, 2a, 3, 4 or 5 is administered orally, intravenously, ure on dialysis. Subcutaneously, transdermally, intraperitoneally, or by inha 0028. In illustrative embodiments, the pulmonary disorder lation. In illustrative embodiments, the kinase receptor is a is associated with abnormal: right ventricular systolic pres receptor tyrosine kinase (RTK), and wherein the RTK is sure (RVSP); pulmonary pressure; cardiac output; right ven platelet derived growth factor receptor (PDGFR). In illustra tive embodiments, the PDGFR is platelet derived growth tricular (RV) hypertrophy; and/or pulmonary arterial (PA) factor receptor-alpha (PDGFR-O.) or platelet derived growth hypertrophy. In illustrative embodiments, the compound of factor receptor-beta (PDGFR-3) or both. In illustrative Structure 1 possesses an ICs of less than 300 nM for the embodiments, the PDGFR is a homodimer or heterodimer kinase receptor. In illustrative embodiments, the kinase selected from PDGFR-CC, PDGFR-BB and PDGFR-CfB, or receptor is platelet derived growth factor receptor-alpha any combination thereof. In illustrative embodiments, the (PDGFR-O.) or platelet derived growth factor receptor-beta inhibition of the PDGFR is effective in treating the pulmo (PDGFR-B) or both, and where the pulmonary disorder is nary disorder, where the pulmonary disorder is pulmonary pulmonary arterial hypertension. In illustrative embodi arterial hypertension (PAH), PAH associated with plexiform ments, the inhibition occurs through a non-covalent interac and/or neointimal lesions, PAH associated with pulmonary tion. In illustrative embodiments, the inhibition occurs fibrosis and/or progressive vaso-degeneration, abnormal through a covalent interaction. fibroblast and/or myofibroblast proliferation, or pulmonary 0029. In one aspect, the present disclosure provides a vascular disorders associated with abnormal endothelial cell method of treating pulmonary arterial hypertension (PAH) in proliferation, or any combination thereof. a Subject, including: modulating the phosphorylation-state of 0025. In illustrative embodiments, the inhibition is a com one or more downstream targets of platelet derived growth bined inhibition of both the PDGFR-a and the PDGFR-B. In factor receptor-alpha (PDGFR-C) or platelet derived growth illustrative embodiments, the inhibition prevents activation of factor receptor-beta (PDGFR-B) or both, where the down both the PDGFR-C. and the PDGFR-B by modulating cognate stream target is any Substrate phosphorylated as a result of the Substrate interactions. In illustrative embodiments, the cog PDGFR-C. and/or the PDGFR-B activation, by administering nate substrate is selected from PDGFAA, PDGFBB and to the Subject a compound of Structure 1, a tautomer, enanti PDGFAB, or any combination thereof. In illustrative embodi omer, isomeror Stereoisomer of the compound, a pharmaceu ments, the pulmonary disorder is selected from pulmonary tically acceptable salt of the compound, tautomer, enanti arterial hypertension (PAH), PAH associated with plexiform omer, isomer or Stereoisomer of the compound, or any and/or neointimal lesions, PAH associated with pulmonary mixtures thereof, where the downstream target is selected fibrosis and/or progressive vasodegeneration, abnormal from the group consisting of AKT, PDGFR, STAT3, ERK1 fibroblast and/or myofibroblast proliferation, and pulmonary and ERK2, or any other downstream target of the PDGFR-C. vascular disorders associated with abnormal endothelial cell and/or the PDGFR-B, and where the compound of Structure 1 proliferation. has the following formula: US 2015/0353527 A1 Dec. 10, 2015
groups, —C(=O)NH(aryl) groups, —C(=O)N(alkyl). Structure 1 groups, —C(=O)N(aryl) groups, —C(=O)N(alkyl)(aryl) R1 groups, —C(=O)C)-alkyl groups, —C(=O)C-aryl groups, R2 —C(=O)-heterocyclyl groups, —C(=O)—O-heterocyclyl 21 R4 groups, —C(=O)NH(heterocyclyl) groups, —C(=O)—N (heterocyclyl) groups, —C(=O)—N(alkyl)(heterocyclyl) . . a groups, —C(=O)—N(aryl) (heterocyclyl) groups, Substi 1S s sk1 tuted and unsubstituted heterocyclylaminoalkyl groups, Sub stituted and unsubstituted hydroxyalkyl groups, Substituted R71 X nx2 X n R5 and unsubstituted alkoxyalkyl groups, Substituted and unsub stituted aryloxyalkyl groups, and Substituted and unsubsti k tuted heterocyclyloxyalkyl groups, Substituted and unsubsti tuted diheterocyclylaminoalkyl, Substituted and 0030 where X is independently selected from C, N, O, S unsubstituted (heterocyclyl)(alkyl)aminoalkyl, substituted or CN: and unsubstituted (heterocyclyl) (aryl)aminoalkyl, Substi 0031) R', R, and R may be the same or different and are tuted and unsubstituted alkoxyalkyl groups, Substituted and independently selected from the group consisting of H, C, N, unsubstituted hydroxyalkyl groups, Substituted and unsubsti O, S, Cl, Br, F, I, CN, NO, -OH, -CH, -CF, tuted aryloxyalkyl groups, and Substituted and unsubstituted —C N–C - groups, —C N–C - groups. (=O)R’ heterocyclyloxyalkyl groups: -(alkyl)(aryl)aminoalkyl groups, N C(=O)R groups, C N C (=O)R groups, —C(=O)-heterocyclyl groups, —C(=O)—O-het groups, substituted and unsubstituted R groups, substituted erocyclyl groups, —C(=O)NH(heterocyclyl) groups, and unsubstituted R groups substituted with one or more of —C(=O)—N (heterocyclyl) groups, —C(=O)—N(alkyl) R. R', and R', substituted and unsubstituted amidinyl (heterocyclyl) groups, —C(=O)—N(aryl) (heterocyclyl) groups, Substituted and unsubstituted guanidinyl groups, Sub groups, Substituted and unsubstituted heterocyclylami stituted and unsubstituted primary, secondary, and tertiary noalkyl groups, Substituted and unsubstituted hydroxyalkyl alkyl groups, Substituted and unsubstituted aryl groups, Sub groups, Substituted and unsubstituted alkoxyalkyl groups, stituted and unsubstituted alkenyl groups, Substituted and Substituted and unsubstituted aryloxyalkyl groups, and Sub unsubstituted alkynyl groups, Substituted and unsubstituted stituted and unsubstituted heterocyclyloxyalkyl groups, heterocyclyl groups, substituted and unsubstituted ami —NH(alkyl) groups, -NH(aryl) groups, —N(alkyl) noalkyl groups, Substituted and unsubstituted alkylami groups, —N(aryl) groups, —N(alkyl)(aryl) groups, NH noalkyl groups, Substituted and unsubstituted dialkylami (heterocyclyl) groups, —N(heterocyclyl)(alkyl) groups, noalkyl groups, Substituted and unsubstituted arylaminoalkyl —N(heterocyclyl)(aryl) groups, —N(heterocyclyl) groups, groups, Substituted and unsubstituted diarylaminoalkyl Substituted and unsubstituted alkyl groups, Substituted and groups, Substituted and unsubstituted (alkyl)(aryl)ami unsubstituted aryl groups, Substituted and unsubstituted noalkyl groups, Substituted and unsubstituted heterocyclyla alkoxy groups, Substituted and unsubstituted aryloxy groups, lkyl groups, Substituted and unsubstituted cyano groups, Sub substituted and unsubstituted heterocyclyl groups, NHOH, stituted and unsubstituted pyrimidinyl groups, Substituted —N(alkyl)OH groups, N(aryl)OH groups, N(alkyl)O- and unsubstituted cyano (aryl) groups, Substituted and unsub alkyl groups, —N(aryl)O-alkyl groups, —N(alkyl)O-aryl stituted cyano (heterocyclyl) groups, and Substituted and groups, and —N(aryl)O-aryl groups; unsubstituted cyano-pyrimidinyl groups; 0033 where the structure of R has the following formula: 0032. R. R. R. and R7, may be the same or different and are independently selected from the group consisting of H, Cl, Br, F, I, —CN, NO. —OH, -CH, —CF, NH, —C=N, -C=N groups, —C N C - groups, —C N C(=O)— groups, —C N C(=O) C F. —C N C(=O)—C—C, substituted and unsubstituted alkyl groups, Substituted and unsubstituted aryl groups, Sub stituted and unsubstituted heterocyclyl groups, alkoxy groups, aryloxy groups, Substituted and unsubstituted hetero cyclylalkyl groups, Substituted and unsubstituted aminoalkyl 0034 and where X is independently selected from C, N, O, groups, Substituted and unsubstituted alkylaminoalkyl S, or -CN: groups, Substituted and unsubstituted dialkylaminoalkyl 0035) R. R', and R' may be the same or different and groups, Substituted and unsubstituted arylaminoalkyl groups, are independently selected from the group consisting of H. C. Substituted and unsubstituted diarylaminoalkyl groups, Sub N, O, S, Cl, Br, F, I, CN, NO, OH, -CH, -CF, stituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, NH, -C(=O) , —C N R', -C=N, C-N C Substituted and unsubstituted alkylamino groups, Substituted (O) C F. —C N C(=O)—C—C, substituted and and unsubstituted arylamino groups, Substituted and unsub unsubstituted alkyl groups, Substituted and unsubstituted aryl stituted dialkylamino groups, Substituted and unsubstituted groups, Substituted and unsubstituted heterocyclyl groups, diarylamino groups, Substituted and unsubstituted (alkyl) —OH, alkoxy groups, aryloxy groups, Substituted and unsub (aryl)amino groups, —C(=O)H, —C(=O)-alkyl groups, stituted heterocyclylalkyl groups, Substituted and unsubsti —C(=O)-aryl groups, —C(=O)C)-alkyl groups, —C(=O) tuted aminoalkyl groups, Substituted and unsubstituted alky O-aryl groups, C(=O)NH2. —C(=O)NH(alkyl) groups, laminoalkyl groups, Substituted and unsubstituted —C(=O)NH(aryl) groups, —C(=O)N(alkyl) groups, dialkylaminoalkyl groups, Substituted and unsubstituted ary —C(=O)-aryl groups, —C(=O)NH2. —C(=O)NH(alkyl) laminoalkyl groups, Substituted and unsubstituted diarylami US 2015/0353527 A1 Dec. 10, 2015 noalkyl groups, Substituted and unsubstituted (alkyl)(aryl) 0039. In illustrative embodiments, the structure of R is aminoalkyl groups, Substituted and unsubstituted alkylamino selected from the Group A structures noted above in the groups, Substituted and unsubstituted arylamino groups, and Summary. In illustrative embodiments, the modulation is a Substituted and unsubstituted dialkylamino groups, Substi decrease of phosphorylated STAT3 to total STAT3, diphos tuted and unsubstituted cyano groups, Substituted and unsub phorylated ERK1 to total ERK1, diphosphorylated ERK2 to stituted pyrimidinyl groups, Substituted and unsubstituted total ERK2, monophosphorylated ERK1 to total ERK1, cyano (aryl) groups, Substituted and unsubstituted cyano (het phosphorylated PDGFR to total PDGFR, or phosphorylated erocyclyl) groups, and Substituted and unsubstituted cyano AKT to total AKT, or any combination thereof, in the subject pyrimidinyl groups; compared to the PSR in the subject before the administering. 0036) R' is selected from the group consisting of In illustrative embodiments, the compound of Structure 1 —C(=O)— groups, —C(=O)—C-groups, —C(=O)— interacts with AKT at residues Thr308 and/or Ser473, or C=C. —S(=O) - groups, —S(=O), C - groups, where the compound of Structure 1 interacts with one or more —S(=O) -C=C- groups, —S(=O) -C=C-CH of the PDGFR-C, PDGFR-B, PDGFR-O.C., PDGFR-B B, and/ —OH, alkoxy groups, aryloxy groups, Substituted and unsub or the PDGFR-O?3 amino acids selected from LYS627, stituted amidinyl groups, Substituted and unsubstituted VAL607, GLU644, MET648, HIS816, LEU809, ASP836, guanidinyl groups, Substituted and unsubstituted primary, CYS814, ILE834, CYS835, PHE937, LYS634, VAL614, secondary, and tertiary alkyl groups, Substituted and unsub GLU651, MET655, HIS824, LEU817, ASP844, CYS822, stituted aryl groups, Substituted and unsubstituted alkenyl ILE842, VAL658, ILE647, HIS816, ARG836, LYS634, groups, Substituted and unsubstituted alkynyl groups, Substi GLU651, ALA632, HIS824, MET655, ARG825, CYS843, tuted and unsubstituted heterocyclyl groups, Substituted and THR874, ARG817, VAL815, LEU651, LEU809, ILE657, unsubstituted aminoalkyl groups, Substituted and unsubsti THR681, ILE654, ARG825, ASP826, LEU658, LEU825, tuted alkylaminoalkyl groups, Substituted and unsubstituted PHE837, LEU658, HIS824, CYS814, ILE654, ASP844, dialkylaminoalkyl groups, Substituted and unsubstituted ary ILE842, and/or CYS843, or any combination thereof. laminoalkyl groups, Substituted and unsubstituted diarylami 0040. In some embodiments, the compound of Structure 1 noalkyl groups, Substituted and unsubstituted (alkyl)(aryl) is a compound selected from the Group C structures noted aminoalkyl groups, Substituted and unsubstituted above in the Summary. In illustrative embodiments, the inhi heterocyclylalkyl groups, Substituted and unsubstituted bition occurs through a non-covalent interaction. In illustra cyano groups, Substituted and unsubstituted pyrimidinyl tive embodiments, the inhibition occurs through a covalent groups, Substituted and unsubstituted cyano(aryl) groups, interaction. In illustrative embodiments, compound of Struc Substituted and unsubstituted cyano(heterocyclyl) groups, ture 1, a tautomer, enantiomer, isomer or stereoisomer of the and Substituted and unsubstituted cyano-pyrimidinyl groups; compound, a pharmaceutically acceptable salt of the com and pound, tautomer, enantiomer, isomer or stereoisomer of the compound, or any mixtures thereof, for treating one or more 0037 where the structure of Q' or Q is selected from the diseases associated with hyperproliferation, neoplasia, hypo group consisting of —CH-OH, -O-CH - C N C plasia, hyperplasia, dysplasia, metaplasia, prosoplasia, des (=O)–C–C, C N C(=O) C F. moplasia, angiogenesis, inflammation, immunological state, metabolism, pulmonary function, and cardiovascular func tion by non-selectively inhibiting a receptor tyrosine kinase o1 o1 (RTK) selected from AKT, c-Kit, and/or PDGFR, where o1 Structure 1 is as follows:
Structure 1 RI
o1 OH 2 Q?, Q?, R 21 R4 1S s - o1 -*n2^n R5 N, and k
0041 where X, R", R°, R s R, R, R, R7 (and R, R, Rio, R', and R', as contained therein), Q' and Q (and Q as contained therein) are selected from XRO3, as noted above. OH I0042. In illustrative embodiments, the structure of R is selected from the Group A structures noted above on the Summary. In some embodiments, the compound is a structure selected from the Group C structures noted above in the Summary. 0038. The entire contents of the foregoing paragraph (i.e., 0043. In illustrative embodiments, the compound of Struc 0019) are hereinafter referred to as “QXR3' ture 1 is administered orally, intravenously, Subcutaneously, US 2015/0353527 A1 Dec. 10, 2015 transdermally, intraperitoneally, or by inhalation. In illustra Imatinib (A). FIG. 2D shows that PDGFBB stimulation of tive embodiments, the disease is selected from the group pAKT(Thr308) was blocked by PK10453 () with an ICs of consisting of cancer, metastatic cancer, HIV, hepatitis, PAH, 0.43 uM compared to 3.25 uM for imatinib (A). FIG. 2E primary PAH, idiopathic PAH, heritable PAH, refractory shows examples of ICWs for PDGFAA and PDGFBB stimu PAH, BMPR2, ALK1, endoglin associated with hereditary lated AKT phosphorylation, PK10453 vs. Imatinib. The sig hemorrhagic telangiectasia, endoglin not associated with nal at 800 nm is color coded green and represents the phos hereditary hemorrhagic telangiectasia, drug-induced PAH, pho-protein specific signal; the signal at 700 nm is color and toxin-induced PAH, PAH associated with systemic scle coded red and represents signal from total AKT. As shown, rosis, and mixed connective tissue disease, pulmonary hyper the 800 and 700nm signals are superimposed (Sp-0.01; p <0. tension, congenital heart disease, hypoxia, chronic hemolytic 001). anemia, newborn persistent pulmonary hypertension, pulmo 0047 FIG. 3 depicts fluorescence images of frozen rat nary veno-occlusive disease (PVOD), pulmonary capillary lung sections (right upper, middle, and lower lobes) after 2 hemangiomatosis (PCH), left heart disease pulmonary hyper min of PK10453 (Structure 2) and IR780 tracer inhalation. tension, systolic dysfunction, diastolic dysfunction, Valvular Image acquisition occurred at 800 nm (green), which is the 2. disease, lung disease, interstitial lung disease, pulmonary of IR780 detection, while image acquisition at 700 nm (red) fibrosis, schistosomiasis, COPD, sleep-disordered breathing, represents tissue autofluorescence. Digital ruler intervals are alveolar hypoVentilation disorders, chronic exposure to high show (1 cm). altitude, developmental abnormalities, chronic thromboem 0048 FIG. 4 shows graphic data for intravenous (IV) and bolic pulmonary hypertension (CTEPH), pulmonary hyper inhaled (INH) PK10453 (Structure 2). FIG. 4A is a pharma tension with unclear multifactorial mechanisms, hematologic cokinetic (PK) graph concerning IV administered PK10453 disorders, myeloproliferative disorders, splenectomy, sys and associated concentrations in the lungs and plasma as a temic disorders, sarcoidosis, pulmonary Langerhans cell his function of time. FIG. 4B is a PK graph concerning INH tiocytosis, lymphangioleimoyomatosis, neurofibromatosis, administered PK10453 and associated levels in the lungs and metabolic disorders, glycogen storage disease, Gaucher dis plasma per time. ease, thyroid disorders, tumor obstruction, fibrosing medias 0049 FIG. 5 depicts the effect of PK10453 (Structure 2) tinitis, and chronic renal failure on dialysis. on right ventricle (RV) systolic pressure and RV hypertrophy 0044. In illustrative embodiments, the salt is a chloride, in the MCT and MCT+PN model systems. FIG.5A is a graph hydrochloride, Sulfate, phosphate, mesylate, bismesylate, showing the effect of PK10453 on RV systolic pressure in the tosylate, lactate, tartrate, malate, bis-acetate, citrate, or bishy MCT model, where C (n-3), V (n=2), D2 (n=6), D4 (n=6), drochloride salt. In illustrative embodiments, the inhibition and D8 (n=5) respectively represent control, vehicle, 2 min occurs through a non-covalent interaction. In some embodi exposure, 4 min exposure, and 8 min exposure times, for two ments, the inhibition occurs through a covalent interaction. In weeks, three times daily. Asterisks (*) indicate p-0.001 and Some embodiments, compound of Structure 1 possesses an section symbols (S) indicate p-0.05. FIG. 5B is a graph ICs of less than 300 nM for the kinase receptor. In illustrative showing the effect of PK10453 on RV hypertrophy in the embodiments, the treatment methods result in one or more of MCT model, where inhalation treatments were initiated three improved exercise capacity, improved functional class, less weeks after administration of MCT. C., D2, D4, and D8 shortness of breath, decreased hospitalization, decreased respectively represent controls, 2, 4, and 8 min exposure need for lung transplantation, decreased need for atrial sep times, for two weeks three times daily. The asterisks (*) tostomy, and increased longevity or overall Survival. In some indicate p-0.001. FIG. 5C is a graph showing the effect of embodiments, the improved exercise capacity is an increased PK10453 on RV systolic pressure (RVSP) in the rat MCT 6 minute walk distance. In suitable embodiments, improved model: comparison of PK10453 to imatinib; if p <0.01. FIG. functional class is an improvement from class IV to class III, SD shows the Lumen/Media ratio in the MCT model of II or I, or an improvement from class III to class II or I, or an PK10453, Imatinib and vehicle: Vehicle (V. n=4): 0.55+0.1; improvement form class II to class I. PK10453 (D8, n=12): 0.94+0.08: Imatinib (I8, n=5): 0.99+0. 07: Sp-0.05, it p-0.01. BRIEF DESCRIPTION OF THE FIGURES 0050 FIG. 6 shows graphs for telemetry studies in the rat 004.5 FIG. 1 shows a graph (PDGFRA: ATP Km app or MCT+PN model. FIG. 6A is a graph showing pulmonary PDGFRB: ATP Km app) depicting ICs concentrations for artery systolic pressure measured over time in ambulatory Imatinib and PK10453 (Structure 2). FIG. 1A shows that the subjects using the MCT+PN model system with PK10453. V ICs of Imatinib against PDGFRC. is 71 nM, while FIG. 1B (n=5) and D4 (n-6) respectively represent vehicle and 4 min shows an ICs for PK10453 against PDGFRC. of 35 nM. FIG. exposure to PK10453 (Structure 2) three times daily. Aster 1C, furthermore, shows that the ICs of Imatinib for PDGFRB isks (*) indicate p-0.001 and section symbols (S) indicate is 607 nM, while FIG.1D shows an ICs for PK10453 against p-0.01. FIG. 6B is a graph showing pulmonary artery systolic PDGFRB of 10.1 nM. pressure measured over time in ambulatory Subjects using the 0046 FIG. 2 shows graphs and images of In Cell Western MCT+PN model system with Imatinib. V-vehicle: (ICW) assays demonstrating the lower ICs of PK10453 I=Imatinib (p=NS). (Structure 2) against PDGFBB stimulated phosphorylation of 0051 FIG. 7 represents graphs relating to hemodynamic AKT at Ser473 and Thr308 compared to Imatinib in human and morphometric analyses in the rat MCT+PN model. FIG. fetal lung fibroblasts. FIGS. 2A-B show that PDGFAA stimu 7A shoes that RV systolic pressure: V (n=9) RVS, 75.7+7.1 lation of p AKT(S473) and pAKTCT308), respectively, in mm Hg, D4 group (n=10) RVS 40.4+2.7 mm Hg, D8 (n=8) HLFs were blocked by PK10453 () and Imatinib (A) with a RVS 43+3.0 mm Hg (p<0.001 V vs. D4 and V vs. D8). FIG. comparable ICso between 0.3-0.6 uM. FIG. 2C shows that 7B shows RV hypertrophy was decreased by treatment with PDGFBB stimulation of p AKT(Ser473) was blocked by PK10453 (Structure 2); (RV+IVS)/LV ratio: V (n=11); D4 PK10453 () with an ICs of 0.13 uM compared to 1.8uM for (n=13); D8 (n=7); *p-0.001, Sp-0.05. FIG.7C shows the rat US 2015/0353527 A1 Dec. 10, 2015
MCT+PN model, the lumen area/media area ratio was greater little signal in the media for pPDGFRB. Signal for pPDGFRB in the D8 (n=5) treated groups compared to PK10453 D4 is noted in peri-vascular cells (upper left FIG. 13C and D), (n-6) and vehicle (n-6); *p-0.0001 D8 vs. V. D8 vs. D4. FIG. and endothelial cells. 7D shows occlusion analyses, which were performed on the 0.058 FIG. 14 shows NanoPro Immunoassays shown for same animal samples used for the lumen/media ratio analysis. the MCT+PN model. FIG. 14A shows p AKT (Thr308) and The occlusion analysis showed a significant decrease in total AKT, with vehicle treatment. FIG. 14B shows p AKT Grade 2 (>50% occlusive) lesions in the D8 group (iip-0.01). (Thr308) and total AKT with PK10453 treatment, while FIG. 0052 FIG. 8 illustrates the effect of PK10453 (Structure 14C shows pAKT (Ser473) and total AKT, with vehicle treat 2) on neointimal lesions in the rat MCT+PN model via 40x ment. FIG. 14D shows pAKT(Ser473) and total AKT with microscopic images of pulmonary arteriole hypertrophy and PK10453 treatment, while FIG. 14E shows that the paKT intraluminal cellular proliferation of PK10453 treated speci (Thr308)/AKT ratio in lung extracts was not significantly mens. FIG. 8A shows a microscope image of neointimal different between the groups (V-vehicle; D4-4 minute expo lesions. FIG. 8B shows an image of PK10453 treated sub sure 3x/day for 2 weeks, D8–8 minute exposure 3x/day for jects. FIG. 8C shows a phosphoPDGFRB (ppDGFRB) stain, two weeks, p=NS). FIG. 14F represents the paKT(Ser473)/ vehicle treated animal, while FIG. 8D shows a pPDGFRB AKT ratio in lung extracts for D8 group vs. vehicle (V, n=5; stain for PK10453 (D8) treated animals. D4, n=4; D8, n=5) Sp-0.05 D8 vs. V. 0053 FIG. 9 is a graph showing lumen area: media area, 0059 FIG. 15 reveals results from experiments employing which is increase in D4 (n-6) and D8 (n=5) treated groups the NanoPro Immunoassay lumogram for pSTAT3 and compared to vehicle (n-6) via MCT+PN model. Symbol (S) STAT3 in the MCT+PN model. FIG. 15A is a graph of the is p=0.032 (D4 vs. V), symbol (; ) is p=0.028 (D8 vs. D4), vehicle treated subjects. FIG. 15B is a graph of the PK10453 and asterisk (*) indicates p=0.00014 (D8 vs. V). (Structure 2) treated subjects. FIG. 15C shows a graph of PK10453 treatment, which decreased pSTAT3/STAT3 in the 0054 FIG. 10 depicts an immunohistochemical evalua lungs of subjects using the MCT+PN model (n=4), where V tion of MCT+PN samples. FIG. 10A shows that pSTAT3 represents vehicle, D4 represents 4 min exposure times three localized to the nuclei of endothelial cells and perivascular times daily, and D8 represent 8 min exposure times for two cells with vehicle treatment. FIG. 10B shows lung pSTAT3 weeks three times daily; 3x/day for two weeks PK10453. nuclear signal from a subject treated with Structure 2. Asterisks (*) p=0.009 and section symbols (S) indicate p=0. 0055 FIG. 11 relates to vehicle treated animals (MCT+PN O24. model) at 40x for immunohisto-chemically stained CSMC 0060 FIG. 16 shows the results from experiments using actin, Trichrome and VWF stains, which showed a mixed the Nanopro Immunoassay lumograms for phosphoERK 1/2 population of endothelial and myofibroblast cells comprising (pERK1/2) and total ERK1/2 in the MCT+PN model. FIG. the neointimal and proliferative lesions in pulmonary arteri 16A shows pERK1/2 in vehicle treated subjects. FIG. 16B oles in Grade 0, 1, and 2 lesions: Grade 0 lesions were char shows pERK1/2 in PK10453 treated subjects. FIG. 16C acterized by early intraluminal endothelial cell proliferation, shows total ERK1/2 and vehicle treated subjects. FIG. 16D and presence of vascular Smooth muscle cells in the media shows total ERK1/2 in PK10453 treated subjects, where (FIG. 11A, C.SMC stain: FIG. 11D, trichrome: FIG. 11G, PK10453 decreased ppERK1/ERK1. FIG. 16E shows VWF). Grade 1-2 lesions had extensive intraluminal myofi ppERK1/ERK1 in subjects as indicated. FIG. 16F shows broblast-like cells, some endothelial cells, and partial fibrosis pERK2/ERK2 as indicated. FIG.16G shows ppERK2/ERK2 of medial layer (FIG. 11B, CSMC; FIG. 11E, trichrome: FIG. as indicated in the lungs. FIG. 16H shows pFRK2/ERK2 as 11 H. VWF). Advanced Grade 2 lesions were characterized by indicated in the lungs. The n=4 for each group, while V extensive intraluminal myofibroblast-like and endothelial represents vehicle, D4 represents 4 min exposure times, three cell proliferation and complete fibrotic replacement of medial times daily, and D8 represents 8 min exposure times of layer (FIG. 11C, CSMC; FIG. 11F, trichrome: FIG. 11I. PK10453 (Structure 2) for two weeks three times daily. Aster VWF). The long arrow points to the intraluminal space with isks (*) p<0.0005: Sp=0.045. proliferative lesions, and the short arrow points to the medial 0061 FIG. 17 is a graphic representation showing the layer of the pulmonary arterioles. effect of imatinib, PK10453 (Structure 2), and PK10571 0056 FIG. 12 shows 40x PDGFR signaling in the rat (Structure 2a) on PDGFAA vs. PDGFBB stimulated phos MCT+PN model. FIGS. 12A-F show PDGFAA in a pulmo phorylation of ERK1 and ERK2 in human fetal lung fibro nary arteriole (A): PDGFBB (B): total PDGFRO. (C); total blasts. See FIG. 17A-D. PDGFRB (D); phosphoPDGFRC (pPDGFRO: E); and 0062 FIG. 18A-D is a graphic representation of PK com pPDGFRB (F). Signal intensity was greater for PDGFBB, pounds: PK10453 (Structure 2), PK10467 (Structure 3), PDGFRB, and especially pPDGFRB compared to PDGFAA, PK10468 (Structure 4), PK10569 (Structure 5) and PK10571 PDGFRC, and pPDGFRC. The pPDGFRB signal was intense (Structure 2a), showing that all PK compounds possess a in a cobblestone pattern in neointimal proliferative and lower ICso concentration compared to imatinib for inhibiting perivascular lesions. Signal intensity was relatively low in PDGFBB stimulated AKT phosphorylation in fetal human vessel media layer. Arrows point to vessel lumens with pro lung fibroblasts. FIG. 18A shows lower ICs of PK10467 liferative lesions (slides are from vehicle treatment). (Structure 3) compared to imatinib, and even lower ICso 0057 FIG. 13 shows a comparison of pPDGFRC. and PK10571 (Structure 2a) vs. PK10467 (Structure 3) and ima pPDGFRB in larger pulmonary arterioles using the rat MCT+ tinib for inhibition of PDGFBB stimulated phosphorylation PN model system. FIGS. 13A and 13B respectively show 20x of AKT in fetal human lung fibroblasts. FIG. 18B shows the and 40x immunohistochemistry for pPDGFRC. signal in the comparison of PK10453 (Structure 2) and PK10571 (Struc media, where the arrow points to a Smooth muscle cell posi ture 2a) with regard to PDGFBB stimulated phosphorylation tive for pPDGFRC. FIGS. 13C and D respectively show at of AKT in fetal human lung fibroblasts. PK10571 (Structure 20x and 40x imaging that, in contrast to above, there was very 2a) has a lower ICs for inhibition of PDGFBB stimulated US 2015/0353527 A1 Dec. 10, 2015
phosphorylation of AKT FIG. 18C shows the comparison of 0070. As used herein, the terms “comparable' or “corre PK10468 (Structure 4) and PK10569 (Structure 5) to ima sponding in the context of comparing two or more samples, tinib; PK10468 (Structure 4) and PK10569 (Structure 5) are responses to treatment, or drugs, refer to the same type of more potent inhibitors of PDGFBB stimulated AKT phos sample, response, treatment, and drug respectively used in the phorylation in fetal human lung fibroblasts. comparison. For example, the phosphorylation state or level 0063 FIG. 19 is a graphic representation of subject body of AKT (pAKT) in a sample can be compared to the phos weight in vehicle administered and PK10453 (Structure 2) phorylation state or level in another sample. In some embodi treated Subjects, where squares indicate vehicle treated ments, comparable samples may be obtained from the same (n=10), triangles indicate PK10453 D4 group (n=10), and individual at different times. In other embodiments, compa diamonds indicate PK10453 D8 group (n-6). rable samples may be obtained from different individuals, 0064 FIG. 20 is a graph representing PAC40 telemetry e.g., a patient and a healthy individual. In general, compa transmitter data from transmitters implanted in the abdominal rable samples are normalized by a common factor for control aorta for monitoring systemic blood pressure for seven days purposes. in ambulatory MCT exposed subjects treated with vehicle 0071. As used herein, the term “composition” refers to a (n-3) or PK10453 (n=3). product with specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, DETAILED DESCRIPTION from combination of the specified ingredients in the specified amountS. 0065. The present disclosure relates to, interalia, a novel 0072. As used herein, the terms “drug.” “compound.” class of compounds which function askinase inhibitors. Like “active agent.” “agent,” “actives.” “pharmaceutical composi wise, methods for using Such compounds in the prevention tion.” “pharmaceutical formulation and “pharmacologi and treatment of disease conditions are disclosed herein. The cally active agent” are used interchangeably and refer to any present disclosure further relates to pharmaceutical formula chemical compound, complex or composition, charged or tions of the compounds, which possess prophylactic and/or uncharged, that is Suitable for administration and that has a therapeutic indications for Subjects in need of kinase inhibi beneficial biological effect, suitably atherapeutic effect in the tors, e.g., patients afflicted with vascular disease, proliferative treatment of a disease or abnormal physiological condition, disorders, cancers, and related diseases or conditions, as fur although the effect may also be prophylactic in nature. The ther detailed below. The definitions of certainterms as used in terms also encompass pharmaceutically acceptable, pharma this specification are provided below. Unless defined other cologically active derivatives of those active agents specifi wise, all technical and Scientific terms used herein generally cally mentioned herein, including, but not limited to, salts, have the same meaning as commonly understood by one of esters, amides, prodrugs, active metabolites, analogs, and the ordinary skill in the art to which this invention belongs. like. When the terms “active agent,” “pharmacologically 0066. As used in this specification and the appended active agent, and “API (active pharmaceutical ingredient) claims, the singular forms “a”, “an and “the' include plural are used, then, or when a particular active agentis specifically referents unless the content clearly dictates otherwise. For identified, it is to be understood that applicants intend to example, reference to “an amino acid' includes a combina include the active agent per se as well as pharmaceutically tion of two or more nucleic acids, and the like. Moreover, as acceptable, pharmacologically active salts, esters, amides, used herein, the following abbreviations have certain mean prodrugs, metabolites, analogs, etc. ings as detailed below. 0073. As used herein, the terms “effective amount’ or 0067. As used herein, “about will be understood by per “pharmaceutically effective amount’ or “therapeutically sons of ordinary skill in the art and will vary to some extent effective amount of a composition, is a quantity Sufficient to depending upon the context in which it is used. If there are achieve a desired therapeutic and/or prophylactic effect, e.g., uses of the term which are not clear to persons of ordinary an amount which results in the prevention of, or a decrease in, skill in the art, given the context in which it is used, “about the symptoms associated with a disease that is being treated. will mean up to plus or minus 10% of the enumerated value. The amount of a composition of the invention administered to 0068. As used herein, the following PK compounds and the subject will depend on the type and severity of the disease structure designations are used interchangeably throughout and on the characteristics of the individual. Such as general the application: PK10453=Structure 2: PK10571=Structure health, age, sex, body weight and tolerance to drugs. It will 2a: PK10467=Structure 3: PK10468=Structure 4; and also depend on the degree, severity and type of disease. The PK10569=Structure 5. skilled artisan will be able to determine appropriate dosages 0069. As used herein, the “administration of an agent or depending on these and other factors. The compositions of the drug, e.g., one or more kinase inhibitor compounds, to a present invention can also be administered in combination Subject or Subjects includes any route of introducing or deliv with one or more additional therapeutic compounds. ering to a subject a compound to perform its intended func 0074 As used herein, the terms “irreversible' or “irrevers tion. Administration can be carried out by any suitable route, ibly' when referring to a kinase inhibitor means an inhibitor including orally, intranasally, by inhalation, parenterally (in of the activity of a kinase, tyrosine kinase, and/or RTK, which travenously, intramuscularly, intraperitoneally, or Subcutane is covalently, i.e., permanently, bound or associated with Such ously), rectally, or topically. Administration includes self a kinase. administration and the administration by another. It is also to 0075. As used herein, the term “neoplastic disease” refers be appreciated that the various modes of treatment or preven to cancers of any kind and origin and precursor stages thereof. tion of medical conditions as described are intended to mean Accordingly, the term "neoplastic disease includes the Sub “substantial, which includes total but also less than total ject matter identified by the terms “neoplasia”, “neoplasm”. treatment or prevention, and where some biologically or “cancer”, “pre-cancer or “tumor.” A neoplastic disease is medically relevant result is achieved. generally manifest by abnormal cell division resulting in an US 2015/0353527 A1 Dec. 10, 2015 abnormal level of a particular cell population. Likewise, undecyl, dodecyl and the like. The phrase also includes because the monoclonal expansion of endothelial cells may branched chain isomers of straight chain alkyl groups, includ refer to a “neoplasm' of the pulmonary arteriolar endothelial ing but not limited to, the following which are provided by cells, PAH is also encompassed within the foregoing terms. way of example: —CH(CH), —CH(CH)(CHCH), The abnormal cell division underlying a neoplastic disease, —CH(CHCH), —C(CH), —C(CH2CH), —CHCH moreover, is typically inherent in the cells and not a normal (CH), —CH2CH(CH)(CHCH), —CHCH(CHCH), physiological response to infection or inflammation. In some —CHC(CH), —CHC(CHCH), —CH(CH)CH(CH) embodiments, neoplastic diseases for diagnosis using meth (CHCH), —CHCH-CH(CH), —CHCH-CH(CH) ods provided herein include carcinoma. (CHCH), —CHCH-CH(CHCH), —CHCHC(CH), 0076. As used herein, the term “non-selective', when - CHCHC(CHCH), —CH(CH)CHCH(CH), —CH referring to a kinase inhibitor or receptor kinase inhibitor, (CH)CH(CH)CH(CH), —CH(CHCH)CH(CH)CH means an inhibitor of the activity of a kinase, tyrosine kinase, (CH)(CHCH), and others. The phrase also includes cyclic domain, and/or RTK, which is not solely specific to a single alkyl groups such as cycloalkyl groups such as cyclopropyl. kinase, receptor, tyrosine kinase, RTK or domain, i.e., a cog cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and nate target, but within the context of inhibiting a single kinase, cyclooctyl and Such rings Substituted with Straight and receptor, tyrosine kinase, RTK, domain, etc., e.g., for branched chain alkyl groups as defined above. The phrase PDGFR, the inhibitor is non-specific with respect to affinities also includes polycyclic alkyl groups such as, but not limited and/or ICso concentrations for the kinase, receptor, tyrosine to, adamantyl norbornyl, and bicyclo2.2.2]octyl and Such kinase, RTK, domain, etc. For example, PK10453 (Structure rings Substituted with straight and branched chain alkyl 2) targets PDGFR, non-selectively, by inhibiting both groups as defined above. Thus, the phrase unsubstituted alkyl PDGFR-Band PDGFR-C. isoforms, but nevertheless may still groups includes primary alkyl groups, secondary alkyl possesses a lower ICs for a receptor isoform, e.g., PDGFR-B. groups, and tertiary alkyl groups. Unsubstituted alkyl groups 0077. As used herein, the term “pharmaceutically accept may be bonded to one or more carbon atom(s), oxygenatom able salt includes a salt with an inorganic base, organic base, (s), nitrogenatom(s), and/or Sulfur atom(s) in the parent com inorganic acid, organic acid, or basic or acidic amino acid. As pound. Preferred unsubstituted alkyl groups include straight salts of inorganic bases, the invention includes, for example, and branched chain alkyl groups and cyclic alkyl groups alkali metals such as sodium or potassium; alkaline earth having 1 to 20 carbon atoms. More preferred such unsubsti metals such as calcium and magnesium or aluminum; and tuted alkyl groups have from 1 to 10 carbonatoms while even ammonia. As salts of organic bases, the invention includes, more preferred such groups have from 1 to 5 carbon atoms. In for example, trimethylamine, triethylamine, pyridine, Some embodiments, unsubstituted alkyl groups include picoline, ethanolamine, diethanolamine, and triethanola straight and branched chain alkyl groups having from 1 to 3 mine. As salts of inorganic acids, the instant invention carbon atoms and include methyl, ethyl, propyl, and —CH includes, for example, hydrochloric acid, hydroboric acid, (CH3)2. nitric acid, Sulfuric acid, and phosphoric acid. As salts of I0081. As used herein, the term “substituted alkyl refers to organic acids, the instant invention includes, for example, an unsubstituted alkyl group as defined above in which one or formic acid, acetic acid, trifluoroacetic acid, fumaric acid, more bonds to a carbon(s) or hydrogen(s) are replaced by a oxalic acid, tartaric acid, maleic acid, lactic acid, citric acid, bond to non-hydrogen and non-carbonatoms such as, but not Succinic acid, malic acid, methanesulfonic acid, benzene limited to, a halogen atom in halides such as F. Cl, Br, and I; Sulfonic acid, and p-toluenesulfonic acid. As Salts of basic an oxygen atom in groups such as hydroxyl groups, alkoxy amino acids, the instant invention includes, for example, argi groups, aryloxy groups, and ester groups; a Sulfur atom in nine, lysine and ornithine. Acidic amino acids include, for groups such as thiol groups, alkyl and aryl Sulfide groups, example, aspartic acid and glutamic acid. Sulfone groups, Sulfonyl groups, and Sulfoxide groups; a 0078. As used herein, the term “reference level refers to a nitrogenatom in groups such as amines, amides, alkylamines, level of a substance which may be of interest for comparative dialkylamines, arylamines, alkylarylamines, diarylamines, purposes. In some embodiments, a reference level may be a N-oxides, imides, and enamines; a silicon atom in groups specified composition dosage as an average of the dose level Such as in trialkylsilyl groups, dialkylarylsilyl groups, alkyl from samples taken from a control Subject. In other embodi diarylsilyl groups, and triarylsilyl groups; and other heteroa ments, the reference level may be the level in the same subject toms in various other groups. Substituted alkyl groups also at a different time, e.g., a time course of administering, Such include groups in which one or more bonds to a carbon(s) or as a level at 2, 4, 6, 8, and 10 minutes (min), etc. hydrogen(s) atom is replaced by a bond to a heteroatom Such 0079. As used herein, the terms “treating” or “treatment” as oxygen in carbonyl, carboxyl, and estergroups; nitrogen in or “alleviation” refer to both therapeutic treatment and pro groups such as imines, OXimes, hydrazones, and nitriles. In phylactic or preventative measures, wherein the objective is Suitable embodiments, Substituted alkyl groups include, to prevent or slow down (lessen) the targeted pathologic con among others, alkyl groups in which one or more bonds to a dition or disorder. A subject is successfully “treated for a carbon or hydrogen atom is/are replaced by one or more disorder if, after receiving a therapeutic agent according to bonds to fluorine atoms. One example of a substituted alkyl the methods of the present invention, the subject shows group is the trifluoromethyl group and other alkyl groups that observable and/or measurable reduction in or absence of one contain the trifluoromethyl group. Other alkyl groups include or more signs and symptoms of a particular disease or con those in which one or more bonds to a carbon or hydrogen dition. atom is replaced by a bond to an oxygen atom such that the 0080. As used herein, the term “unsubstituted alkyl refers Substituted alkyl group contains a hydroxyl, alkoxy, aryloxy to alkyl groups that do not contain heteroatoms. Thus the group, or heterocyclyloxy group. Still other alkyl groups phrase includes straight chain alkyl groups such as methyl, include alkyl groups that have an amine, alkylamine, dialky ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl lamine, arylamine, (alkyl)(aryl)amine, diarylamine, hetero US 2015/0353527 A1 Dec. 10, 2015
cyclylamine, (alkyl)(heterocyclyl)amine, (aryl)(heterocycly I0088 As used herein, the term “unsubstituted aralkyl l)amine, or diheterocyclylamine group. refers to unsubstituted alkyl groups as defined above in which 0082. As used herein, the term “unsubstituted aryl refers a hydrogen or carbon bond of the unsubstituted alkyl group is to aryl groups that do not contain heteroatoms. Thus the term replaced with a bond to an aryl group as defined above. For includes, but is not limited to, groups such as phenyl, biphe example, methyl (-CH-) is an unsubstituted alkyl group. If nyl, anthracenyl, naphthenyl by way of example. Although a hydrogenatom of the methyl group is replaced by a bond to the phrase “unsubstituted aryl includes groups containing a phenyl group, Such as if the carbon of the methyl were condensed rings such as naphthalene, it does not include aryl bonded to a carbon of benzene, then the compound is an groups that have other groups such as alkyl or halo groups unsubstituted aralkyl group, i.e., a benzyl group. Thus, the bonded to one of the ring members, as aryl groups such as term includes, but is not limited to, groups such as benzyl, tolyl are considered herein to be substituted aryl groups as diphenylmethyl, and 1-phenylethyl( CH(CH5)(CH)), described below. Unsubstituted aryl groups may be bonded to among others. one or more carbon, oxygen, nitrogen, and/or Sulfur atom(s). I0089. As used herein, the term “substituted aralkyl has 0083. As used herein, the term “substituted aryl group' the same meaning with respect to unsubstituted aralkyl has the same meaning with respect to unsubstituted aryl groups that Substituted aryl groups had with respect to unsub groups that Substituted alkyl groups had with respect to stituted aryl groups. However, a Substituted aralkyl group also unsubstituted alkyl groups. However, a Substituted aryl group includes groups in which a carbon or hydrogen bond of the also includes aryl groups in which one of the aromatic car alkyl part of the group is replaced by a bond to a non-carbon bons is bonded to one of the non-carbon or non-hydrogen or a non-hydrogen atom. Non-limiting examples of Substi atoms described above and also includes aryl groups in which tuted aralkyl groups include —CH2C(=O)(CHs), and one or more aromatic carbons of the aryl group is bonded to —CH2(2-methylphenyl), among others. a substituted and/or unsubstituted alkyl, alkenyl, or alkynyl 0090. As used herein, the term “unsubstituted heterocy group as defined herein. This includes bonding arrangements clyl refers to both aromatic and nonaromatic ring com in which two carbonatoms of an aryl group are bonded to two pounds including monocyclic, bicyclic, and polycyclic ring atoms of an alkyl, alkenyl, or alkynyl group to define a fused compounds Such as, but not limited to, quinuclidyl, contain ring system (e.g. dihydronaphthyl or tetrahydronaphthyl). ing 3 or more ring members of which one or more is a Thus, the term “substituted aryl' includes, but is not limited heteroatom such as, but not limited to, N, O, and S. Examples to, tolyl and hydroxyphenyl, among others. of heterocyclyl groups include, but are not limited to: unsat 0084 As used herein, the term “unsubstituted alkenyl' urated 3 to 8 membered rings containing 1 to 4 nitrogenatoms refers to straight and branched chain and cyclic groups such Such as, but not limited to pyrrolyl, pyrrolinyl, imidazolyl, as those described with respect to unsubstituted alkyl groups pyrazolyl pyridinyl, dihydropyridinyl, pyrimidyl, pyrazinyl, as defined above, except that at least one double bond exists pyridazinyl, triazolyl, e.g., 4H-1,2,4-triazolyl, 1H-1,2,3-tria between two carbon atoms. Non-limiting examples include Zolyl, 2H-1,2,3-triazolyl etc., tetrazolyl, e.g., 1H-tetrazolyl, vinyl, -CH=C(H)(CH), -CH=C(CH), C(CH)—C 2H tetrazolyl, etc.); saturated 3 to 8 membered rings contain (H), —C(CH)=C(H)(CH), —C(CHCH)—CH, cyclo ing 1 to 4 nitrogen atoms such as, but not limited to, pyrro hexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, penta lidinyl, imidazolidinyl, piperidinyl, piperazinyl; condensed dienyl, and hexadienyl among others. unsaturated heterocyclic groups containing 1 to 4 nitrogen 0085. As used herein, the term “substituted alkenyl has atoms Such as, but not limited to, indolyl, isoindolyl, indoli the same meaning with respect to unsubstituted alkenyl nyl, indolizinyl, benzimidazolyl, quinolyl, isoquinolyl, inda groups that Substituted alkyl groups had with respect to Zolyl, benzotriazolyl; unsaturated 3 to 8 membered rings unsubstituted alkyl groups. A substituted alkenyl group containing 1 to 2 oxygenatoms and 1 to 3 nitrogenatoms Such includes alkenyl groups in which a non-carbon or non-hydro as, but not limited to, oxazolyl, isoxazolyl, oxadiazolyl, e.g., gen atom is bonded to a carbon double bonded to another 1,2,4-oxadiazolyl, 1.3,4-oxadiazolyl, 1,2,5-oxadiazolyl, etc.; carbon and those in which one of the non-carbon/non-hydro saturated 3 to 8 membered rings containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms such as, but not limited to, gen atoms is bonded to a carbon not involved in a carbon morpholinyl; unsaturated condensed heterocyclic groups double bond. containing 1 to 2 oxygenatoms and 1 to 3 nitrogenatoms, for I0086. As used herein, the term “unsubstituted alkynyl example, benzoxazolyl, benzoxadiazolyl, benzoxazinyl, e.g. refers to straight and branched chain groups such as those 2H-1,4-benzoxazinyl, etc.); unsaturated 3 to 8 membered described with respect to unsubstituted alkyl groups as rings containing 1 to 3 Sulfur atoms and 1 to 3 nitrogenatoms defined above, except that at least one triple bond exists Such as, but not limited to, thiazolyl, isothiazolyl, thiadiaz between two carbon atoms. Examples include, but are not olyl, e.g.,1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadia limited to, -C=C(H), -C=C(CH), -C=C(CHCH), Zolyl, 1,2,5-thiadiazolyl, etc.; saturated 3 to 8 membered —C(H)C=C(H), C(H),C=C(CH), and C(H)C=C rings containing 1 to 2 Sulfur atoms and 1 to 3 nitrogenatoms (CH2CH), among others. Such as, but not limited to, thiazolodinyl; Saturated and unsat 0087 As used herein, the term “substituted alkynyl has urated 3 to 8 membered rings containing 1 to 2 sulfur atoms the same meaning with respect to unsubstituted alkynyl such as, but not limited to, thienyl, dihydrodithiinyl, dihy groups that Substituted alkyl groups had with respect to drodithionyl, tetrahydrothiophene, tetrahydrothiopyran: unsubstituted alkyl groups. A substituted alkynyl group unsaturated condensed heterocyclic rings containing 1 to 2 includes alkynyl groups in which a non-carbon or non-hydro Sulfur atoms and 1 to 3 nitrogenatoms such as, but not limited gen atom is bonded to a carbon triple bonded to another to, benzothiazolyl, benzothiadiazolyl, benzothiazinyl (e.g. carbon and those in which a non-carbon or non-hydrogen 2H-1,4-benzothiazinyl, etc.), dihydrobenzothiazinyl, e.g., atom is bonded to a carbon not involved in a carbon triple 2H-3,4-dihydrobenzothiazinyl, etc., unsaturated 3 to 8 mem bond. bered rings containing oxygenatoms Such as, but not limited US 2015/0353527 A1 Dec. 10, 2015 to furyl; unsaturated condensed heterocyclic rings containing with respect to unsubstituted aralkyl groups. However, a Sub 1 to 2 oxygen atoms such as benzodioxolyl, e.g., 1,3-benzo stituted heterocyclylalkyl group also includes groups in dioxoyl, etc.; unsaturated 3 to 8 membered rings containing which a non-hydrogen atom is bonded to a heteroatom in the an oxygen atom and 1 to 2 sulfur atoms such as, but not heterocyclyl group of the heterocyclylalkyl group Such as, but limited to, dihydrooxathiinyl; saturated 3 to 8 membered not limited to, a nitrogen atom in the piperidine ring of a rings containing 1 to 2 oxygen atoms and 1 to 2 sulfur atoms piperidinylalkyl group. In addition, a Substituted heterocycly Such as 1,4-oxathiane; unsaturated condensed rings contain lalkyl group also includes groups in which a carbon bond or a ing 1 to 2 Sulfur atoms such as benzothienyl, benzodithiinyl: hydrogen bond of the alkyl part of the group is replaced by a and unsaturated condensed heterocyclic rings containing an bond to a substituted and unsubstituted aryl or substituted and oxygenatom and 1 to 2 oxygenatoms Such as benzoxathiinyl. unsubstituted aralkyl group. Heterocyclyl group also include those described above in 0094. As used herein, the term “unsubstituted alkylami which one or more S atoms in the ring is double-bonded to noalkyl refers to an unsubstituted alkyl group as defined one or two oxygen atoms (sulfoxides and Sulfones). For above in which a carbon or hydrogen bond is replaced by a example, heterocyclyl groups include tetrahydrothiophene bond to a nitrogenatom that is bonded to a hydrogenatom and oxide and tetrahydrothiophene 1,1-dioxide. Preferred hetero an unsubstituted alkyl group as defined above. For example, cyclyl groups contain 5 or 6 ring members. More preferred methyl (-CH-) is an unsubstituted alkyl group. If a hydrogen heterocyclyl groups include morpholine, piperazine, piperi atom of the methyl group is replaced by a bond to a nitrogen dine, pyrrolidine, imidazole, pyrazole, 1,2,3-triazole, 1,2,4- atom that is bonded to a hydrogen atom and an ethyl group, triazole, tetrazole, thiophene, thiomorpholine, thiomorpho then the resulting compound is CH N(H)(CHCH) which line in which the Satom of the thiomorpholine is bonded to is an unsubstituted alkylaminoalkyl group. one or more O atoms, pyrrole, homopiperazine, oxazolidin 0.095 As used herein, the term “substituted alkylami 2-one, pyrrolidin-2-one, oxazole, quinuclidine, thiazole, noalkyl refers to an unsubstituted alkylaminoalkyl group as isoxazole, furan, and tetrahydrofuran. defined above except where one or more bonds to a carbon or 0091. As used herein, the term “substituted heterocyclyl hydrogen atom in one or both of the alkyl groups is replaced refers to an unsubstituted heterocyclyl group as defined above by a bond to a non-carbon or non-hydrogenatomas described in which one or more of the ring members are bonded to a above with respect to substituted alkyl groups except that the non-hydrogen atom Such as described above with respect to bond to the nitrogenatom in all alkylaminoalkyl groups does Substituted alkyl groups and Substituted aryl groups. not by itself qualify all alkylaminoalkyl groups as being Sub Examples, include, but are not limited to, 2-methylbenzimi stituted. dazolyl, 5-methylbenzimidazolyl, 5-chlorobenzthiazolyl, N-alkylpiperazinyl groups such as 1-methylpiperazinyl, pip 0096. As used herein, the term “unsubstituted dialkylami erazine-N-oxide, N-alkyl piperazine N-oxides, 2-phenoxy noalkyl refers to an unsubstituted alkyl group as defined thiophene, and 2-chloropyridinyl among others. In addition, above in which a carbon bond or hydrogen bond is replaced Substituted heterocyclyl groups also include heterocyclyl by a bond to a nitrogen atom which is bonded to two other groups in which the bond to the non-hydrogenatom is a bond unsubstituted alkyl groups as defined above. to a carbon atom that is part of a Substituted and unsubstituted 0097. As used herein, the term “substituted dialkylami aryl, substituted and unsubstituted aralkyl, or unsubstituted noalkyl refers to an unsubstituted dialkylaminoalkyl group heterocyclyl group. Examples include but are not limited to as defined above in which one or more bonds to a carbon or 1-benzylpiperidinyl, 3-phenythiomorpholinyl, 3-(pynolidin hydrogenatom in one or more of the alkyl groups is replaced 1-yl)-pyrrolidinyl, and 4-(piperidin-1-yl)-piperidinyl. by a bond to a non-carbon and non-hydrogen atom as Groups such as N-alkyl Substituted piperazine groups such as described with respect to substituted alkyl groups. The bond N-methyl piperazine, Substituted morpholine groups, and to the nitrogenatom in all dialkylaminoalkyl groups does not piperazine N-oxide groups such as piperazine N-oxide and itself qualify all dialkylaminoalkyl groups as Substituted. N-alkyl piperazine N-oxides are examples of some substi 0098. As used herein, the term “unsubstituted alkoxy” tuted heterocyclyl groups. Groups such as Substituted pipera refers to a hydroxyl group (-OH) in which the bond to the Zine groups such as N-alkyl Substituted piperazine groups hydrogen atom is replaced by a bond to a carbon atom of an such as N-methyl piperazine and the like, substituted mor otherwise unsubstituted alkyl group as defined above. As pholine groups, and N-oxide groups are examples of some used herein, the term “substituted alkoxy' refers to a substituted heterocyclyl groups that are suited for various “R” hydroxyl group (-OH) in which the bond to the hydrogen groups. atom is replaced by a bond to a carbon atom of an otherwise 0092. As used herein, the term “unsubstituted heterocy Substituted alkyl group as defined above. clylalkyl refers to unsubstituted alkyl groups as defined 0099. As used herein, the term “unsubstituted heterocy above in which a hydrogen or carbon bond of the unsubsti clyloxy' refers to a hydroxyl group (-OH) in which the bond tuted alkyl group is replaced with a bond to a heterocyclyl to the hydrogenatom is replaced by a bond to a ring atom of group as defined above. For example, methyl (—CH) is an an otherwise unsubstituted heterocyclyl group as defined unsubstituted alkyl group. If a hydrogen atom of the methyl above. As used herein, the term “substituted heterocyclyloxy” group is replaced by a bond to a heterocyclyl group. Such as if refers to a hydroxyl group (-OH) in which the bond to the the carbon of the methyl were bonded to carbon 2 of pyridine hydrogen atom is replaced by a bond to a ring atom of an (one of the carbons bonded to the N of the pyridine) or otherwise substituted heterocyclyl group as defined above. As carbons 3 or 4 of the pyridine, then the compound is an used herein, the term “unsubstituted heterocyclyloxyalkyl unsubstituted heterocyclylalkyl group. refers to an unsubstituted alkyl group as defined above in 0093. As used herein, the term “substituted heterocycly which a carbon bond or hydrogen bond is replaced by an lalkyl has the same meaning with respect to unsubstituted oxygen bond, which is bonded to an unsubstituted heterocy heterocyclylalkyl groups that Substituted aralkyl groups had clyl group. US 2015/0353527 A1 Dec. 10, 2015
0100. As used herein, the term “substituted heterocycly 0106. As used herein, the term “substituted alkylami loxyalkyl refers to an unsubstituted heterocyclyloxyalkyl noalkoxy' refers to unsubstituted alkylaminoalkoxy groups group as defined above in which a bond to a carbon or hydro as defined above in which a bond to a carbon or hydrogen gen group of the alkyl group of the heterocyclyloxyalkyl atom of the alkyl group bonded to the oxygen atom which is group is bonded to a non-carbon and non-hydrogen atom as bonded to the parent compound is replaced by one or more described above with respect to substituted alkyl groups or in bonds to a non-carbon and non-hydrogen atoms as discussed which the heterocyclyl group of the heterocyclyloxyalkyl above with respect to substituted alkyl groups and/or if the group is a Substituted heterocyclyl group as defined above. hydrogen bonded to the amino group is bonded to a non 0101. As used herein, the term “unsubstituted heterocy carbon and non-hydrogen atom and/or if the alkyl group clylalkoxy' refers to an unsubstituted alkyl group as defined above in which a carbon bond or hydrogen bond is replaced bonded to the nitrogen of the amine is bonded to a non-carbon by a bond to an oxygen atom which is bonded to the parent and non-hydrogen atom as described above with respect to compound, and in which another carbon or hydrogen bond of Substituted alkyl groups. The presence of the amine and the unsubstituted alkyl group is bonded to an unsubstituted alkoxy functionality in all alkylaminoalkoxy groups does not heterocyclyl group as defined above. As used herein, the term by itself qualify all Such groups as Substituted alkylami “substituted heterocyclylalkoxy' refers to an unsubstituted noalkoxy groups. heterocyclylalkoxy group as defined above in which a bond to 0107 As used herein, the term “unsubstituted dialkylami a carbon or hydrogen group of the alkyl group of the hetero noalkoxy' refers to an unsubstituted alkyl group as defined cyclylalkoxy group is bonded to a non-carbon and non-hy above in which a carbon or hydrogen bond is replaced by a drogen atom as described above with respect to substituted bond to an oxygen atom which is bonded to the parent com alkyl groups or in which the heterocyclyl group of the hetero cyclylalkoxy group is a Substituted heterocyclyl group as pound and in which another carbon or hydrogen bond of the defined above. Further, a substituted heterocyclylalkoxy unsubstituted alkyl group is bonded to a nitrogenatom which group also includes groups in which a carbon bond or a is bonded to two other similar or different unsubstituted alkyl hydrogen bond to the alkyl moiety of the group may be groups as defined above. substituted with one or more additional substituted and 0108. As used herein, the term “substituted dialkylami unsubstituted heterocycles. noalkoxy' refers to an unsubstituted dialkylaminoalkoxy 0102. As used herein, the term “unsubstituted arylami group as defined above in which a bond to a carbon or hydro noalkyl” refers to an unsubstituted alkyl group as defined genatom of the alkyl group bonded to the oxygenatom which above in which a carbon bond or hydrogen bond is replaced is bonded to the parent compound is replaced by one or more by a bond to a nitrogen atom which is bonded to at least one bonds to a non-carbon and non-hydrogen atoms as discussed unsubstituted aryl group as defined above. above with respect to substituted alkyl groups and/or if one or 0103) As used herein, the term “substituted arylami more of the alkyl groups bonded to the nitrogen of the amine noalkyl refers to an unsubstituted arylaminoalkyl group as is bonded to a non-carbon and non-hydrogen atom as defined above except where either the alkyl group of the described above with respect to substituted alkyl groups. The arylaminoalkyl group is a Substituted alkyl group as defined presence of the amine and alkoxy functionality in all dialky above or the aryl group of the arylaminoalkyl group is a laminoalkoxy groups does not by itself qualify all Such Substituted aryl group except that the bonds to the nitrogen groups as Substituted dialkylaminoalkoxy groups. atominallarylaminoalkyl groups does not by itself qualify all 0109 As used herein, the term “protected with respect to arylaminoalkyl groups as being Substituted. However, Substi hydroxyl groups, amine groups, and Sulfhydryl groups refers tuted arylaminoalkyl groups does include groups in which the to forms of these functionalities which are protected from hydrogenbonded to the nitrogenatom of the group is replaced undesirable reaction with a protecting group known to those with a non-carbon and non-hydrogen atom. skilled in the art such as those set forth in Protective Groups 0104. As used herein, the term “unsubstituted heterocy in Organic Synthesis, Greene, T. W.; Wuts, P. G. M., John clylaminoalkyl refers to an unsubstituted alkyl group as Wiley & Sons, New York, N.Y., (3rd Edition, 1999), which defined above in which a carbon or hydrogen bond is replaced can be added or removed using the procedures set forth by a bond to a nitrogen atom which is bonded to at least one therein. Examples of protected hydroxyl groups include, but unsubstituted heterocyclyl group as defined above. As used are not limited to, silyl ethers such as those obtained by herein, the term “substituted heterocyclylaminoalkyl refers reaction of a hydroxyl group with a reagent such as, but not to unsubstituted heterocyclylaminoalkyl groups as defined limited to, t-butyldimethyl-chlorosilane, trimethylchlorosi above in which the heterocyclyl group is a substituted hetero lane, triisopropylchlorosilane, triethylchlorosilane; Substi cyclyl group as defined above and/or the alkyl group is a tuted methyl and ethyl ethers such as, e.g., methoxymethyl substituted alkyl group as defined above. The bonds to the ether, methylthiomethyl ether, benzyloxymethyl ether, t-bu nitrogen atom in all heterocyclylaminoalkyl groups does not toxymethyl ether, 2-methoxyethoxymethyl ether, tetrahydro by itself qualify all heterocyclylaminoalkyl groups as being pyranyl ethers, 1-ethoxyethyl ether, allyl ether, benzyl ether; substituted. esters such as, but not limited to, benzoylformate, formate, 0105. As used herein, the term “unsubstituted alkylami acetate, trichloroacetate, and trifluoracetate. Non-limiting noalkoxy' refers to an unsubstituted alkyl group as defined examples of protected amine groups include amides Such as, above in which a carbon or hydrogen bond is replaced by a formamide, acetamide, trifluoroacetamide, and benzamide; bond to an oxygen atom which is bonded to the parent com imides, such as phthalimide, and dithiosuccinimide; and oth pound and in which another carbon or hydrogen bond of the ers. Non-limiting examples of protected Sulfhydryl groups unsubstituted alkyl group is bonded to a nitrogenatom which include thioethers such as S-benzylthioether, and 5-4-picolyl is bonded to a hydrogen atom and an unsubstituted alkyl thioether; substituted S-methyl derivatives such as hemithio. group as defined above. dithio and aminothio acetals, among others. US 2015/0353527 A1 Dec. 10, 2015
Overview characteristic of human disease, e.g., PAH. To this end, for 0110 Various compounds have been found useful in treat example, the pathologic signature of PAH consists of concen ing certain diseases such as, e.g., cancer. For example, tric and plexiform lesions in Small precapillary pulmonary GleevecR) (imatinib mesylate or “imatinib) is a compound arterioles. See Cooletal. (1997); and Tuder et al., “Plexiform that has shown efficacy in treating chronic myeloid leukemia lesion in severe pulmonary hypertension: association with (CML) and gastrointestinal stromal tumors (GIST). Other glomeruloid lesion.” Am J Pathol 159:382-383 (2001). Con experimental drugs include Sorafenib and PNU-166196 for centric lesions arise from the proliferation of neointimal cells, the respective treatment of renal cell carcinoma and leukemia. which occlude the vessel lumen. It has been reported that Although significant advances have been made in the devel these concentric obstructive neointimal lesions are composed opment of pharmaceutical compositions for treating certain of myofibroblasts and/or endothelial cells. See, e.g., Yi et al., cancers; new compounds, compositions, methods of treat Am J Respir Crit Care Med 162:1577-86 (2000). ment, and model systems for developing drugs are required 0114. In addition, perivascular infiltrates, consisting of T for preventing and/or treating cancer and other diseases, e.g., cells, B cells, and macrophages, have been found in plexo pulmonary-vascular disease such as pulmonary arterial genic PAH. See Sakagami, “In vivo, in vitro and ex vivo hypertension (PAH). In particular, platelet derived growth models to assess pulmonary absorption and disposition of factor (PDGF) receptor tyrosine kinases are an attractive inhaled therapeutics for systemic delivery.” Adv. Drug Deliv therapeutic target for PAH. The PDGF signaling pathway is Rev 58:1030-1060 (2006). Plexiform lesions, moreover, are activated in human idiopathic PAH (iPAH) and in animal characterized by disorganized vascular channels that stain for models of the disease. For example, PDGFA, PDGFB, PDG endothelial cell markers, and Such lesions in lung Samples FRC. and PDGFRB mRNA expression is increased in small from patients with idiopathic and/or primary PAH consist of pulmonary arteries from patients with iPAH compared to a monoclonal expansion of endothelial cells. Lee et al., control Subjects, and Western blot analysis shows a signifi "Monoclonal endothelial cell proliferation is present in pri cant increase in protein expression of PDGFRB in PAH lungs. mary but not secondary pulmonary hypertension.” J. Clin 0111. The migration of PASMCs is inhibited by imatinib, Invest 101:927-934 (1998). As such, PAH of this type is a PDGFRO inhibitor. Imatinib also decreases RVSP and essentially a "cancer of pulmonary arteriolar endothelial improved survival in the rat MCT model of PAH. In several cells (see id.), at least because in the initial or early stages of case reports of patients with refractory PAH, a favorable the disease, an acute apoptotic loss of normal endothelial cells response to imatinib has been observed. See Ghofrani et al., may result in the emergence and clonal expansion of apopto “Imatinib in pulmonary arterial hypertension patients with sis resistantendothelial cells. Lee etal. (1998). The neoplastic inadequate response to established therapy.” Am J Respir Crit process associated with PAH provides for not only kinase Care Med. Vol. 182:1171-7 (2010). The IMPRES trial, which inhibitor treatment of PAH, but also the development of new examined the effect of imatinib in patients with severe PAH, compounds, compositions, and methods, via MCT+PN showed an improvement in the six minute walk distance and model determinations, with Superior efficacy, potency and a in cardiopulmonary hemodynamics. However, orally admin broader spectrum of inhibition compared to previously gen istered imatinib may be associated with systemic side effects erated kinase inhibitors using inferior model systems, which including gastrointestinal distress and bone marrow Suppres may possess a narrow selectivity for RTK inhibition, for the sion. See Paniagua et al., “Imatinib for the treatment of rheu treatment of neoplastic disease. Drug-kinase homology mod matic diseases.” Nat Clin Pract Rheumatol; Vol 3:190-1 eling ensures that Such inhibitors, including, for example, (2007). To improve the therapeutic window, i.e., increase non-selective and irreversibly derivatives thereof, target vul efficacy and decrease systemic side-effects, the present inven nerable kinase domains for optimal efficacy, as further tors employed inhalation delivery of kinase inhibitors for described below. PAH. Compound Synthesis 0112 Imatinib, moreover, was developed using an in vivo murine MCT model system, which is an imperfect system 0.115. In one aspect, the present disclosure provides for the concerning preclinical drug candidate efficacy assessment at synthesis of Structure I compounds, which are readily Syn least because it is unreliable with respect to expressing certain thesized using the procedures described in the following sec human disease phenotypes, e.g., the development of neointi tions and as disclosed in WO 2008/058341, which is hereby mal and/or plexiform lesions associated with PAH. Cool et incorporated by reference in its entirety and for all purposes al., “Pathogenesis and evolution of plexiform lesions in pull as if fully set forth herein. Compounds of Structure I, more monary hypertension associated with Scleroderma and over, are typically prepared from starting materials, such as, human immunodeficiency virus infection.” Hum Pathol. e.g., dihaloheterocycle. The first step is a nucleophilic aro 28:434–442 (1997). Therefore, examining the effects of matic Substitution to generate a monoamino-monohalo inter kinase inhibitors in more aggressive models presenting mediate. The nucleophilic aromatic Substitution is typically human disease phenotypes is essential for more accurately carried out by addition of a primary or secondary amine to the reflecting the pathology of the human disease and, conse di-halogenated heterocycle in a solvent Such as ethanol, iso quently, the development of the next generation of com propanol, tert-butanol, dioxane, THF, DMF, ethoxyethanol, pounds and compositions for effectively treating human dis toluene or Xylene. The reaction typically occurs at elevated CaSC. temperature in the presence of excess amine or a non-nucleo 0113. The present inventors have employed such a model, philic base Such as triethylamine or diisopropylethylamine, or while further comparing the present compounds and thera an inorganic base Such as potassium carbonate or Sodium pies to imatinib. As further detailed below, the present inven carbonate. tors performed efficacy studies using a murine monocrotaline 0116. Alternatively, the amino substituent may be intro (MCT) plus pneumonectomy (PN) model system (MCT+ duced through a transition metal catalyzed amination reac PN). This model imparts neointimal and/or plexiform lesions tion. Typical catalysts for Such transformations include US 2015/0353527 A1 Dec. 10, 2015
Pd(OAc)/P(t-Bu), Pd(dba)/BINAP and Pd(OAc)/BI tion with a suitably functionalized coupling partner to replace NAP. These reactions are typically carried out in solvents the halogenatom with an alternative moiety. Typical coupling Such as toluene or dioxane, in the presence of bases such as partners are organoboronic acids or esters. See, e.g., Miyaura caesium carbonate or sodium or potassium tert-butoxide at and Suzuki, Chem Rev. 952457 (1995); Stille, Chem., Int. Ed. temperatures ranging from room temperature to reflux. See, Engl 25, 508 (1986); Kumada et al., Org. Synth. Coll. Vol.6, e.g., Hartwig and Angew, Chem. Int. Ed 37,2046 (1998). The 407 (1998); and: Negishi, J. Organomet. Chem. 653, 34 amines employed in the first step of the synthesis of these (2002) for Suzuki coupling, organostannanes, Stille cou compounds are obtained commercially or are prepared using pling, Grignard reagents, Kumada coupling, organozinc spe methods well known to those skilled in the art. O-alkylben cies, and Negishi coupling, respectively. The Suzuki coupling Zylamines, moreover, may be prepared through reduction of is the preferred coupling method and is typically performed in oXimes. Typical reductants include lithium aluminium a solvent such as DME, THF, DMF, ethanol, propanol, tolu hydride, hydrogen gas in the presence of palladium on char ene, or 1,4-dioxane in the presence of a base Such as potas coal catalyst, Zn in the presence of hydrochloric acid, sodium sium carbonate, lithium hydroxide, caesium carbonate, borohydride in the presence of a Lewis acid such as TiCb, Sodium hydroxide, potassium fluoride or potassium phos ZrCU, NiCl, and MoO, or sodium borohydride with phate. The reaction may be carried out at elevated tempera Amberlyst H15 ion exchange resin and LiCl, C-Alkylbenzy tures and the palladium catalyst employed may be selected lamines may also be prepared by reductive amination of the from Pd(PPh), Pd(OAc), PdCl(dppf), Pd(dba)/P(t- corresponding ketones. A classical method for Such a trans Bu). formation is the Leuckart-Wallach reaction, though catalytic 0119 The monoamino-monochloro intermediate may conditions (HCONH, CH-)5CRhCl) or other proce also be subjected to a second nucleophilic aromatic Substitu dures, e.g., NHOAc, Na(CN)BH) are also used. C.-Alkyl tion reaction using similar conditions to those outlined above. benzylamines may also be prepared from the corresponding Those skilled in the art will appreciate that the order of the a-alkylbenzyl alcohols. Such methods include derivatisation reactions described for the syntheses above may be changed of the hydroxyl as a mesylate or tosylate and displacement in certain circumstances and that certain functionalities may with a nitrogen nucleophile, Such as phthalimide or azide need to be derivatized, L. e., protected, in certain instances for which is converted to the primary amine using conventional the reactions described above to proceed with reasonable synthetic methods; or, displacement of the hydroxyl with a yield and efficiency. The types of protecting functionality are suitable nitrogen nucleophile under Mitsunobu-like condi well-known to those skilled in the art. The products formed tions. C.-Alkylbenzyl alcohols can be prepared by reduction from the reaction sequences described above may be further of the corresponding ketones with a reducing agent Such as derivatized using techniques well known to those skilled in Sodium borohydride in a solvent Such as methanol. Alterna the art. The leaving group may be any Suitable known type tively, O-alkylbenzyl alcohols can be obtained through addi such as those disclosed in March, Advanced Organic Chem tion of an alkyl metal species (such as a Grignard reagent) to istry: Reactions, Mechanisms and Structure.” 4th Ed. pp. 352 a benzaldehyde derivative, which is typically performed at 7, John Wiley & Sons, NY (1992). In some embodiments, the room temperature or below in solvents such as tetrahydrofu leaving group is a halogen, e.g., chlorine. ran. C.-Alkyl benzylamines of high optical purity may be prepared from chiral O-alkylbenzyl alcohols using the meth Kinases ods outlined above. The chiral O-alkylbenzyl alcohols may I0120 Protein kinases are a family of enzymes that cata be obtained through chiral reduction of the corresponding lyze the phosphorylation of specific residues in proteins. Such ketones. enzymes are generally categorized into three groups, those 0117 The monoamino-monohalo intermediate formed which preferentially phosphorylate serine and/or threonine from the dihaloheterocycle and the amine described above, residues, those which preferentially phosphorylate tyrosine may then be further functionalized. For example, where the residues, and those which phosphorylate both tyrosine and amine Substituent bears an additional functional group, this Ser/Thr residues. Protein kinases are therefore key elements functional group may be derivatized or functionalized using in signal transduction pathways responsible for transducing methods well-known to those skilled in the art. For example, extracellular signals, including the action of cytokines on a free primary amino group could be further functionalized to their receptors, to the nuclei, triggering various biological an amide, Sulphonamide or urea functionality, or could be events. The many roles of protein kinases in normal cell alkylated to generate a secondary or tertiary amine derivative. physiology include cell cycle control including proliferation, Preferable methods for the formation of an amide include differentiation, metabolism, apoptosis, cell mobility, mitoge coupling the amine with a carboxylic acid using coupling nesis, transcription, translation and other signaling processes. reagents such as dicyclohexylcarbodiimide, 1-(3-dimethy I0121 Platelet derived growth factor receptor kinase laminopropyl)-3-ethylcarbodiimide, diisopropylcarbodiim (PDGFR) is one type of RTK. The sequence of PDGFR can be ide or carbonyldiimidazole in solvents such as dichlo found in GenBank, accession number NM-002609 (mRNA) romethane, tetrahydrofuran or 1,4-dioxane. Alternatively, the and NP-002600 (protein) and has been described, at least, in acid component may be activated by conversion to an acid Matsui, et al., “Isolation of a novel receptor cDNA establishes chloride (using thionyl chloride, oxalyl chloride, bis(trichlo the existence of two PDGF receptor genes’ Science romethyl)carbonate or cyanuric chloride) or to mixed anhy 243(4892):800-804 (1989); Claesson-Welsh, L. etal. “cDNA dride species (using, for example, t-butyl chloroformate or cloning and expression of a human platelet-derived growth isopropyl chloroformate) or to active ester intermediates factor (PDGF) receptor specific for B-chain-containing (such as N-hydroxysuccinimidyl, pentafluorophenyl or p-ni PDGF molecules' Mol. Cell. Biol. 8(8):3476-3486 (1988); trophenyl esters) prior to amine reaction. and Gronwald, et al. PNAS. 85(10):3435-3439 (1988). 0118. The monoamino-monochloro intermediate may I0122) Moreover, PDGFR’s cognate binding ligand, then be reacted in a palladium mediated cross-coupling reac PDGF, is a strong mitogenic factor for cells of mesenchymal US 2015/0353527 A1 Dec. 10, 2015
origin such as fibroblasts, Smooth muscle cells, and glial cells. and apoptosis. The Ras-Raf-MEK-ERK signaling cascade PDGF is a 32 kDa protein heterodimer usually composed of controlling cell proliferation has been well studied but the two polypeptide chains, A and B, linked by disulfide bonds. In mechanisms involved in ERK 1/2-mediated cell death are addition to the PDGF AB heterodimer, two homodimeric largely unknown. ERK 1/2 translocates to the nucleus, but can forms of PDGF exist (AA and BB). During blood clotting and also remain in the cytosol. Cytosolic retention of ERK1/2 platelet adhesion, the PDGF is released from granules at sites denies access to the transcription factor Substrates that are of injured blood vessels, suggesting that PDGF may have a responsible for the mitogenic response. In addition, cytosolic role in the repair of blood vessels. PDGF may stimulate ERK 1/2, besides inhibiting survival and proliferative signals migration of arterial Smooth muscle cells from the medial to in the nucleus, potentiates the catalytic activity of some the intimal layer of the artery where the muscle cells may proapoptotic proteins such as DAP kinase in the cytoplasm. proliferate. The cellular proliferation induced by all isoforms Studies that further define the function of cytosolic ERK1/2 of PDGF is mediated by ligand binding to the PDGF receptor. and its cytosolic substrates that enhance cell death will be The PDGF receptor belongs to the class III tyrosine kinase essential to harness this pathway for developing effective family and consists of two receptor Subtypes, termed type A treatments for cancer and chronic inflammatory diseases. (or type alpha), and type B (or type beta), as detailed above. (0.126 STAT3 is a member of the STAT protein family, Other members of the PDGF receptor family include CSF-IR, which typical function in response to cytokines and growth cKIT and FLT3. The two PDGF receptor isoforms may be factors. STAT family members are phosphorylated by the distinguished by their markedly different ligand binding receptor associated kinases, and then form homo- or het specificities. PDGFB receptor binds only B-chain (isoforms erodimers that translocate to the cell nucleus where they act as BB and AB), while PDGFU receptor can bind all forms of transcription activators. STAT3 is activated through phospho PDGF (isoforms containing A and/or B chain). With the rylation in response to various cytokines and growth factors importance of PDGF-related processes to proliferation of including IFNs, EGF, IL5, IL6, HGF, LIF and BMP2. This endothelial cells and vascular Smooth muscle, there are a protein mediates the expression of a variety of genes in range of pathogenic processes that PDGFRIB kinase inhibi response to cell stimuli, and thus plays a key role in many tors are useful for, e.g., disease prevention and treatment. cellular processes Such as cell growth and apoptosis. The 0123 PDGF expression has been shown in a number of small GTPase Racl has been shown to bind and regulate the different solid tumors, from glioblastomas to prostate carci activity of this protein, while PIAS3 has been shown to inhibit nomas. In these various tumor types, the biological role of STAT3. PDGF signaling can vary from autocrine stimulation of can 0127 AKT (also known as PKB) is involved in the regu cer cell growth to more Subtle paracrine interactions involv lation of metabolism, cell Survival, motility, transcription and ing adjacent stroma and angiogenesis. Therefore, inhibiting cell-cycle progression. AKT belongs to the AGC subfamily of the PDGFR kinase activity with small molecules may inter the protein kinase Superfamily, which consists of more than fere with tumor growth, angiogenesis, diseases with neoplas 500 members in humans. The AKT subfamily comprises tic etiologies, immunological and inflammatory diseases, three mammalian isoforms, Akt1, Akt2, and Akt3, which are hyperproliferative diseases including cancer and diseases products of distinct genes and share a conserved structure that involving neo-angiogenesis, renal and kidney diseases, bone includes three functional domains: an N-terminal pleckstrin remodeling diseases, metabolic diseases, vascular diseases, homology (PH) domain, a central kinase domain, and a C-ter and pulmonary vascular diseases such as, e.g., PAH. Other minal regulatory domain containing the hydrophobic motif diseases mediated by PDGF, and thus involving its cognate (HM) phosphorylation site FXXF(S/T)Y. receptors, include, for example, restenosis, including coro nary restenosis after angioplasty, atherectomy, or other inva Kinase Inhibitors sive methods of plaque removal, and renal or peripheral artery I0128. In one aspect, the present disclosure provides com restenosis after the same procedures; vascular proliferative pounds and methods of inhibiting a kinase, e.g., a tyrosine phenomena and fibrosis associated with other forms of acute kinase. Such as a RTK, in a Subject and/or a method of treating injury Such as pulmonary fibrosis associated with adult res a biological condition mediated by, or associated with, a piratory distress syndrome, renal fibrosis associated with kinase, e.g., a tyrosine kinase. Such as a RTK, in a Subject. In nephritis, coronary Stenosis associated with Kawasake's dis Some embodiments, the kinase is Cdc2 kinase, AKT, c-Kit, ease and vascular narrowings associated with other arteritides c-ABL, ERK1/2, STAT3, p.60src, VEGFR3, PDGFRC, PDG Such as Takayasha's disease; prevention of narrowings in vein FRB, FGFR3, PDGFR-Co., PDGFR-BB, PDGFR-CfB, FLT-3, grafts; prevention of narrowings due to accelerated Smooth Fyn, Lck, Tie-2, GSK-3, Cdk2, Cdk4, MEK1, NEK-2, muscle cell migration and proliferation in transplanted CHK2, CK1e, Raf, CHK1, Rsk2, FMS (CSF-IR), KDR, organs, and other fibrotic processes, such as Scleroderma and EphA2, EphA3, EphA8, FLT1, FLT4, HCK, PTKS, RET, myofibrosis and inhibition of tumor cell proliferation. SYK, DDR1, DDR2 and PAR-1. Likewise, the kinase is a 0124 c-Kit is another receptor tyrosine kinase belonging tyrosine kinase. Such as, e.g., Cdc2 kinase, c-Kit, c-ABL, to PDGF Receptor family and is normally expressed in p60src, VEGFR3, PDGFRC, PDGFRB, FGFR3, PDGFR hematopoietic progenitor, mast and germ cells. c-kit expres C.C., PDGFR-B B, PDGFR-O?3, FLT-3, Fyn, Lck, and/or Tie-2, sion has been implicated in a number of cancers including in some embodiments. The methods include administering to mast cell leukemia, germ cell tumors, Small-cell lung carci the subject a compound of Structure I, a tautomer of the noma, GIST, acute myelogenous leukemia (AML), neuro compound, a pharmaceutically acceptable salt of the com blastoma, melanoma, ovarian carcinoma, breast carcinoma. pound, a pharmaceutically acceptable salt of the tautomer, or Smolich et al., Blood, 97(5) 1413-21. mixtures thereof. 0.125 Extracellular signal-regulated protein kinases 1 and I0129. Previously, various indolyl substituted compounds 2 (ERK 1/2) are members of the mitogen-activated protein are shown to inhibit one or more kinases, as disclosed in WO (MAP) kinase super family that can mediate cell proliferation 01/29025, WO 01/62251, and WO 01/62252. Likewise, vari US 2015/0353527 A1 Dec. 10, 2015
ous benzimidazolyl compounds have recently been disclosed adducts. Chloroacetamide is shown as an example of an elec in WOO1/28993. Such compounds are reported to be capable trophile which can form a covalent bond with a cysteine of inhibiting, modulating, and/or regulating signal transduc residue. The general term “warhead' is used to mean an tion of both receptor-type and non-receptor tyrosine kinases. electrophilic trap for forming a covalent bond between the Some of the disclosed compounds contain a quinolone frag inhibitor and the targeted protein kinase. Chloroacetamide as ment bonded to the indolyl or benzimidazolyl group. The an electrophile may be too reactive to have clinical utility and synthesis of 4-hydroxy quinolone and 4-hydroxy quinoline may have toxicity for this reason. Leproult et al. (2011) nev derivatives has also been reported. For example, Ukrainets et ertheless Suggest that less then optimal positioning of the al. have disclosed the synthesis of 3-(benzimidazol-2-yl)-4- electrophile could explain why a covalent bond may not form hydroxy-2-oxo-1,2-dihydroquinoline. Ukrainets et al., Tet. with other less reactive warheads. Lett. 42, 7747-48 (1995) has also disclosed the synthesis, I0133. The present disclosure provides for, inter alio, dis anticonvulsive and antithyroid activity of other 4-hydroxy tinct warhead positioning on RTK receptor inhibitors. In quinolones and thioanalogs such as 1H-2-oxo-3-(2-benzimi some embodiments, electrophiles other than those described dazolyl)-4-hydoxyquinoline. Ukrainets et al., Khimiya by Leproult et al. (2011), were employed for increased effi Geterotsiklicheskikh Soedinii, 1, 105-108 (1993). Other cacy. See Barf et al. (2012) and Oballa et al., “A generally compounds, moreover, such as, for example, 4-Amino-5- applicable method for assessing the electrophilicity and reac fluoro-3-5-(4-methylpiperazin-1-yl)-1H-benzimidazol-2- tivity of diverse nitrile-containing compounds.” Bioorg Med ylquinolin-2(1H)-one has been described as an orally bio Chem Lett 17:998-1002 (2007) (describing nitrile-containing available benzimidazole-quinolinone that exhibits inhibition electrophiles). Furthermore, Diller et al., J Med Chem of receptor tyrosine kinases that drive both endothelial and 46.4638-4647 (2003) reported a homology model of the tumor cell proliferation. The inhibitory effect was shown on PDGFB receptor based on VEGFR2 (55% homology). nine tyrosine kinases, FGFR1, FGFR3, VEGFR1, VEGFR2, 0.134 Molecular docking was previously employed by the VEGFR3, PDGFRP. c-Kit, p60src, and FLT-3, as disclosed in inventors with respect to one aspect of the present invention WO 2005/047244. However, this compound does not signifi by using homology models of RTK, based on homologous cantly inhibit EGFR family kinases or insulin receptor structures, e.g., PDGFC. and PDGFB receptor homology to kinases at pharmaceutically acceptable doses. c-Kit is 59% and 63%, respectively. In some embodiments, 0130. Moreover, 4-(4-methylpiperazin-1-ylmethyl)-N- the introduction of various electrophiles in a variety of posi 4-methyl-3-(4-pyridin-3-yl)pyrimidin-2-ylamino)phenyl tions with respect to a RTK inhibitor, e.g., PDGFR inhibitor, benzamide (imatinib), as disclosed in US 2006/0154936, scaffold provided the bases for further biochemical analyses. inhibits PDGFRC. and 13 kinases, Abl, DDR, and c-KIT, as To this end, the spatial orientation of the inhibitor warheads, described in US 2011/0190313. Paragraph 0117 of US relative to the target cysteine residues, can be analyzed to 2011/01903 13, however, indicates that although imatinib calculate the free energy of binding and estimated K. In some appeared safe and well tolerated over a 6 month period, the embodiments, compounds with the lowest free energy of primary efficacy parameter (6MWD) did not improve in binding and closest proximity of the warhead to a cysteine patients randomized to imatinib compared with placebo, residue impart irreversible non-selective RTK inhibitors. despite significant improvement in secondary endpoints. A 0.135 Accordingly, the present disclosure provides com continuing need therefore exists for compounds that inhibit, pounds of Structure 1, the enantiomer, isomer or Stereoisomer kinases, e.g., tyrosine kinases, such as RTKs, at least because of the compound, a pharmaceutically acceptable salt of the of previous limitations, resistant disease phenotypes, and the compound, tautomer, enantiomer, isomer or stereoisomer of need for more effective kinase, e.g., RTK, inhibition, as fur the compound, or any mixtures thereof, which covalently ther detailed below. See US 2008/02684.60. interact with a receptor tyrosine kinase (RTK), such as, for 0131 Furthermore, the small molecules that were example, PDGFR or c-Kit or both. In some embodiments, the reported in Frey et al. (1998) were shown to irreversibly PDGFR is selected from the group consisting of PDGFR-O. inhibit epidermal growth factor receptor (EGFR) by PDGFR-B, PDGFR-O.C., PDGFR-B B, and PDGFR-of as covalently interacting with the receptor, while alkylating a demonstrated via homology modeling. cysteine residue in the ATP binding pocket of the molecule. Indeed, Leproult et al., “Cysteine Mapping in Conformation Pharmaceutical Compositions ally Distinct Kinase Nucleotide Binding Sites: Application to 0.136. In one aspect, the present disclosure provides phar the Design of Selective Covalent Inhibitors. J. Med. Chem. maceutical compositions which include at least one of the 54, 1347-1355 (2011), discloses that one approach to design compounds of Structure 1 and a pharmaceutically acceptable ing irreversible inhibitors is to exploit the nucleophilicity of a carrier. The compositions of the present invention may con cysteine thiol group present in the target protein via system tain other therapeutic agents as described below, and may be atic analysis of cysteine residues present in the nucleotide formulated, for example, by employing conventional Solid or binding site of kinases. Such an approach can facilitate irre liquid vehicles or diluents, as well as pharmaceutical addi versible inhibition even when taking into consideration the tives of a type appropriate to the mode of desired administra different kinase conformations and therefore improve dosing tion, for example, excipients, binders, preservatives, stabiliz and toxicity. Seeid. ers, flavors, etc., according to techniques such as those well 0132) The cysteine mapping in Leproult et al. (2011) dem known in the art of pharmaceutical formulation. onstrate that kinases are potential targets for selective cova 0.137 Pharmaceutical compositions are typically formu lent inhibitors. An example is shown of the kinase inhibitor lated to be compatible with its intended route of administra imatinib to which a chloroacetamide group is added in the tion. Examples of routes of administration include parenteral para position of the benzene ring. Peptide inhibitor adduct (e.g., intravenous, intradermal, intraperitoneal or Subcutane formation was shown for both Kit and PDGFU receptors. Id. ous), oral, inhalation, transdermal (topical), intraocular, ion However, other compounds failed to show similar covalent tophoretic, and transmucosal administration. Solutions or US 2015/0353527 A1 Dec. 10, 2015
Suspensions used for parenteral, intradermal, or Subcutaneous ALEC). These surfactant formulations are administered via application can include the following components: a sterile airway instillation (i.e., after intubation) or intratracheally. diluent such as water for injection, Saline solution, fixed oils, 0.141. As a further alternative, the agents of the present polyethylene glycols, glycerine, propylene glycol or other invention may be administered to the airways in the form of synthetic solvents; antibacterial agents such as benzyl alcohol an inhalable powder. The powder formulation may include or methyl parabens; antioxidants such as ascorbic acid or physiologically acceptable excipients such as monosaccha Sodium bisulfite; chelating agents such as ethylenediamine rides (e.g. glucose or arabinose), disaccharides (e.g. lactose, tetraacetic acid; buffers such as acetates, citrates or phos saccharose and maltose), oligo- and polysaccharides (e.g. phates and agents for the adjustment of tonicity Such as dextrane), polyalcohols (e.g. Sorbitol, mannitol. Xylitol), Salts sodium chloride or dextrose. The pH can be adjusted with (e.g. sodium chloride, calcium carbonate) or mixtures of acids or bases, such as hydrochloric acid or sodium hydrox these excipients with one another. Preferably, mono- or dis ide. The parenteral preparation can be enclosed in ampoules, accharides are used, while the use of lactose or glucose is disposable Syringes or multiple dose vials made of glass or preferred, particularly, but not exclusively, in hydrate form. plastic. For convenience of the patient or treating physician, 0142. Within the scope of the inhalable powders according the dosing formulation can be provided in a kit containing all to the invention the excipients have a maximum average necessary equipment for a treatment course. particle size of up to 250 um, preferably between 10 and 150 0.138. The compounds of the present disclosure are admin um, most preferably between 15 and 80 Lum. It may sometimes istered by any Suitable means, for example, orally, Such as in seem appropriate to add finer excipient fractions with an the form of tablets, capsules, granules or powders; Sublin average particle size of 1 to 9 um to the excipients mentioned gually; buccally; parenterally, Such as by Subcutaneous, intra above. These finer excipients are also selected from the group venous, intramuscular, intra(trans)dermal, or intracisternal of possible excipients listed hereinbefore. Finally, in order to injection or infusion techniques, e.g., as sterile injectable prepare the inhalable powders according to the invention, aqueous or non-aqueous solutions or Suspensions, nasally micronised formulations, preferably with an average particle Such as by inhalation spray or insufflation, topically, such as size of 0.5 to 10 um is added to the excipient mixture. Pro in the form of a cream or ointment ocularly in the form of a cesses for producing the inhalable powders according to the Solution or Suspension, vaginally in the form of pessaries, invention by grinding and micronizing and by finally mixing tampons or creams, or rectally such as in the form of Supposi the ingredients together are known from the prior art. tories, in unit dosage formulations containing nontoxic, phar 0143. In formulations intended for administration to the maceutically acceptable vehicles or diluents. The compounds respiratory tract, including intranasal formulations, the active may, for example, be administered in a form Suitable for compound is typically configured to have a small particle immediate release or extended release. Immediate release or size, e.g., approximately 5 microns or less, via micronisation extended release may be achieved by the use of suitable techniques and the like. Sustained release formulations of the pharmaceutical compositions comprising the present com active compound are employed in some embodiments. The pounds, or, for extended release, by the use of devices such as active compound, in some embodiments, is administered by Subcutaneous implants or osmotic pumps. oral inhalation as a free-flow powder via inhaler. 0139 For administration to the respiratory tract, e.g., inha 0144. The pharmaceutical composition and method of the lation, including intranasal administration, the active com present disclosure further include additional therapeutically pound may be administered by any of the methods and for active compounds (second agents), as noted herein and/or mulations employed in the art for administration to the known in the art, which are typically employed for treating respiratory tract. Thus, the active compound may be admin one or more pathological conditions in concert with the com istered in the form of e.g., a solution, Suspension, or as a dry positions comprising compounds of Structure 1 of the present powder. The agents according to this aspect of the present disclosure. The combination of therapeutic agents acts Syn invention may also be administered directly to the airways in ergistically to effect the treatment or prevention of the various the form of an aerosol. For use as aerosols, the compounds of diseases, disorders, and/or conditions described herein. Such the present invention in Solution or Suspension may be pack second agents, include, but are not limited to, of prostanoids, aged in a pressurized aerosol container together with Suitable endothelin antagonists, cytoplasmic kinase inhibitors, recep propellants, for example, hydrocarbon propellants like pro tor kinase inhibitors, endothelin receptor antagonists, e.g., pane, butane, or isobutane with conventional adjuvants. The ambrisentan, bosentan, and sitaxsentan, PDES (PDE-V) materials of the present invention also may be administered in inhibitors, e.g., sildenafil, tadalafil, and Vardenafil, calcium a non-pressurized form such as in a nebulizer or atomizer. channel blockers, e.g., amlodipine, felodipine, varepamil, dil 0140. The propellant-driven inhalation aerosols which tiazem, and menthol, prostacyclin, treprostinil, iloprost, bera may be used according to the invention may also contain other prost, nitric oxide, oxygen, heparin, warfarin, diuretics, ingredients such as co-solvents, stabilizers, Surfactants, anti digoxin, cyclosporins, e.g., cyclosporin A, CTLA4-Ig, anti oxidants, lubricants and pH adjusters. The propellant-driven bodies such as ICAM-3, anti-IL-2 receptor (Anti-Tac), anti inhalation aerosols according to the invention which may be CD45RB, anti-CD2, anti-CD3 (OKT-3), anti-CD4, anti used according to the invention may be administered using CD80, anti-CD86, agents blocking the interaction between inhalers known in the art, e.g., metered dose inhalers. As CD40 and gp39, such as antibodies specific for CD40 and/or another alternative, the agents of the present invention may be gp39, i.e., CD 154, fusion proteins constructed from CD40 administered to the airways in the form of a lung Surfactant and gp39 (CD40 1 g and CD8gp39), inhibitors, such as formulation. The lung Surfactant formulation can include nuclear translocation inhibitors, of NF-kappa B function, exogenous lung Surfactant formulations (e.g., InfasurfR (For Such as deoxyspergualin (DSG), cholesterol biosynthesis est Laboratories), Survanta R (Ross Products), and Curo inhibitors such as HMG CoA reductase inhibitors (lovastatin surfR (DEY. California, USA) or synthetic lung surfactant and simvastatin), non-steroidal anti-inflammatory drugs formulations (e.g., ExoSurfR (GlaxoWellcome Inc.) and (NSAIDs) such as ibuprofen, aspirin, acetaminophen, US 2015/0353527 A1 Dec. 10, 2015 20 leflunomide, deoxyspergualin, cyclooxygenase inhibitors about 0.01 to 250 mg/kg per day, from about 0.05 to 100 Such as celecoxib, Steroids such as prednisolone or dexam mg/kg per day, or from about 0.1 to 50 mg/kg per day. Within ethasone, gold compounds, beta-agonists Such as salbutamol. this range, in Some embodiments, the dosage is from about LABAS Such as Salmeterol, leukotriene antagonists such as 0.05 to 0.5, 0.5 to 5 or 5 to 50 mg/kg per day. For oral montelukast, antiproliferative agents such as methotrexate, administration, the compositions are provided in the form of FK506 (tacrolimus, Prograf), mycophenolate mofetil, cyto tablets containing 1.0 to 1000 mg of the active ingredient, toxic drugs such as azathioprine, VP-16, etoposide, fludara including, but not limited to, 1, 5, 10, 15, 20, 25, 50, 75, 100, bine, doxorubin, adriamycin, amsacrine, camptothecin, cyt 150, 200, 250, 300, 400, 500, 600, 750, 800, 900, and 1000 arabine, gemcitabine, fluorodeoxyuridine, melphalan and mg of the active ingredient. The dosage may be selected, for cyclophosphamide, antimetabolites such as methotrexate, example, to any dose within any of these ranges, for thera topoisomerase inhibitors such as camptothecin, DNA alkyla peutic efficacy and/or symptomatic adjustment of the dosage tors such as cisplatin, kinase inhibitors such as Sorafenib, to the Subject being treated. In some embodiments, the com microtubule poisons such as paclitaxel, TNF-Cinhibitors pounds of the present disclosure are administered by inhala such as tenidap, anti-TNF antibodies or soluble TNF receptor, tion as described in, e.g., U.S. Pat. No. 8.257,741, U.S. Pat. hydroxy urea and rapamycin (sirolimus or Rapamune) or No. 8,263,128, WO 2010/132827, WO 2010/102066, WO derivatives thereof. 2012/040502, WO 2012/031129, and/or WO 2010/102065, 0145 The compounds of the invention may also be pre from 1 to 20, 1 to 15, 1 to 10, 1 to 5, 1 to 4, or 1 to 3 times daily, pared as salts which are pharmaceutically acceptable, but it or once or twice per day. In some embodiments, the com will be appreciated that non-pharmaceutically acceptable pounds of the present disclosure are administered from 1 to 5 salts also fall within the scope of the present disclosure at least times daily. to the extent that such salts are useful as intermediates in the 0.147. In some embodiments, the unit dose is sufficient to preparation of pharmaceutically acceptable salts. Examples provide one or more of: (a) a C of about 1 to 5000 ng/mL of pharmaceutically acceptable salts include, but are not lim of the compound In a subjects plasma or a C of about 1 to ited to, Sulfates, phosphates, mesylates, bismesylates, tosy 5000 ng/mL of the compound In the subjects blood when it lates, lactates, tartrates, malates, bis-acetates, citrates, bishy is administered to the subject; and (b) about 1 to 5000 ng/mL drochloride salts, salts of pharmaceutically acceptable of the compound in a subject’s plasma 24 h after administra cations such as sodium, potassium, lithium, calcium, magne tion or about 1 to 5000 ng/mL of the compound in the sub sium, ammonium and alkylammonium; acid addition salts of ject’s blood 24 h after administration to the subject. pharmaceutically acceptable inorganic acids such as hydro 0148. Thetherapeutically effective amount of a compound chloric, orthophosphoric, Sulfuric, phosphoric, nitric, car of Structure 1, the tautomer of the compound, enantiomer, bonic, boric, Sulfamic and hydrobromic acids; or salts of isomer or stereoisomer of the compound, a pharmaceutically pharmaceutically acceptable organic acids such as acetic, acceptable salt of the compound, tautomer, enantiomer, iso propionic, butyric, tartaric, maleic, hydroxymaleic, fumaric, mer or Stereoisomer of the compound, or any mixtures citric, lactic, mucic, gluconic, benzoic. Succinic, oxalic, phe thereof, is not associated with adverse side effects, in some nylacetic, methanesulfonic, trihalomethanesulfonic, toluene embodiments. Such adverse side effects include, but are not Sulfonic, benzenesulfonic, isethionic, Salicylic, Sulphanilic, limited to, decreased lung function, increased or decreased aspartic, glutamic, edetic, Stearic, palmitic, oleic, lauric, pan systemic blood pressure, immunocompromised, bone mar tothenic, tannic, ascorbic, Valeric and orotic acids. Salts of row Suppression, anemia, hypoxia, in the Subject compared to amine groups may also comprise quaternary ammonium salts the Subject prior to the administering. in which the amino nitrogen atom carries a suitable organic group Such as an alkyl, alkenyl, alkynyl or aralkyl moiety. The Prevention and Treatment of Disease salts may be formed by conventional means, such as by react 0149. In one aspect, the present disclosure provides a com ing the free base form of the compound with one or more pound of Structure 1, a tautomer of the compound, enanti equivalents of the appropriate acid in a solvent or medium in omer, isomeror Stereoisomer of the compound, a pharmaceu which the salt is insoluble, or in a solvent such as water which tically acceptable salt of the compound, tautomer, is removed in vacuo or by freeze drying or by exchanging the enantiomer, isomer or stereoisomer of the compound, or any anions of an existing salt for another anion on a suitable ion mixtures thereof for treating one or more diseases, where exchange resin. In some embodiments, the salt is a Sulfate, Structure 1 is described herein. phosphate, mesylate, bismesylate, tosylate, lactate, tartrate, 0150. The present disclosure accordingly provides com malate, bis-acetate, citrate, or bishydrochloride salt. pounds, compositions, and methods of inhibiting kinases, 0146 In some embodiments, the compounds of the e.g., tyrosine kinases, and methods of treating biological con present disclosure are administered in a therapeutically effec ditions mediated by, or associated with, such kinases. For tive amount. Such an administration imparts that a compound example, the present disclosure provides methods of inhibit of Structure 1 will elicit a response associated with, e.g., cells, ing one or more kinases, such as, e.g., cell division cycle 2 tissues, fluids, of a subject being sought by the clinician. In kinase (Cdc2 kinase), c-Kit, c-ABL, p60src, AKT, VEGFR3, the treatment or prevention of conditions mediated by, or PDGFRC, PDGFRB, PDGFR-O.C., PDGFR-B B, PDGFR-O?3, associated with, kinase inhibition, e.g., RTK inhibition, an FGFR3, FLT-3, FYN oncogene kinase related to SRC, FGR, appropriate dosage level is administered. In some embodi YES (Fyn), lymphocyte-specific protein tyrosine kinase ments, from about 0.01 to 500 mg/kg of subject body weight (Lck), tyrosine kinase with Ig and EGF homology domains per day is administered in single or multiple doses. In accord, (Tie-2), FMS (CSF-IR), KDR, EphA2, EphA3, EphA8, dosage levels are from about 0.1 to about 250 mg/kg per day FLT1, FLT4, HCK, PTKS, RET, SYK, DDR1, DDR2, gly in some embodiments, while in other embodiments from cogen synthase kinase 3 (GSK-3), cyclindependent kinase 2 about 0.5 to about 100 mg/kg per day is administered to the (Cdk2), cyclin dependent kinase 4 (Cdk4). MEK1, NEK-2, subject. Suitable dosage levels include, for example, from CHK2, CK1e, Raf, checkpoint kinase 1 (CHK1), ribosomal US 2015/0353527 A1 Dec. 10, 2015
S6 kinase 2 (Rsk2), and PAR-1. In particular, compounds, disease, Gaucher disease, thyroid disorders, tumoral obstruc compositions, and methods of inhibiting tyrosine kinases, tion, fibrosing mediastinitis, and chronic renal failure on Such as, e.g., cell division cycle 2 kinase (Cdc2 kinase), dialysis; and diseases Such as pulmonary hypertension, con ERK 1/2, STAT3, AKT, c-Kit, c-ABL, p60src, VEGFR3, genital heart disease, hypoxia, chronic hemolytic anemia, PDGFRC, PDGFRB, PDGFR-O.C., PDGFR-B|B, PDGFR-O?3, newborn persistent pulmonary hypertension, pulmonary FGFR3, FLT-3, FYN oncogene kinase related to SRC, FGR, veno-occlusive disease (PVOD), pulmonary capillary YES (Fyn), lymphocyte-specific protein tyrosine kinase hemangiomatosis (PCH), left heart disease pulmonary hyper (Lck), tyrosine kinase with Ig and EGF homology domains tension, systolic dysfunction, diastolic dysfunction, Valvular (Tie-2), FMS (CSF-IR), KDR, EphA2, EphA3, EphA8, disease, lung disease, interstitial lung disease, pulmonary FLT1, FLT4, HCK, PTK5, RET, SYK, DDR1, and DDR2. In fibrosis, Schistosomiasis, chronic obstructive pulmonary dis Some embodiments, the tyrosine kinase is a receptor tyrosine ease (COPD), sleep-disordered breathing, alveolar hypoven kinase (RTK), such as, e.g., PDGFR, PDGFR-O.C., PDGFR tilation disorders, chronic exposure to high altitude, develop BB, PDGFR-62 B, or c-Kit, or combinations thereof, are pro mental abnormalities, chronic thromboembolic pulmonary vided. hypertension (CTEPH), pulmonary hypertension with 0151. The present disclosure also provides compounds, unclear multifactorial mechanisms, hematologic disorders, compositions, and methods of treating biological conditions myeloproliferative disorders, splenectomy, systemic disor mediated by, or associated with, kinases, e.g., tyrosine ders, sarcoidosis, pulmonary Langerhans cell histiocytosis, kinases, including Cdc2kinase, c-Kit, AKT, c-ABL, ERK1/2, lymphangioleimoyomatosis, neurofibromatosis, vasculitis, STAT3, p.60src, VEGFR3, PDGFRC, PDGFRB, FGFR3, metabolic disorders, glycogen storage disease, Gaucher dis PDGFR-Co., PDGFR-BB, PDGFR-O?3, FLT-3, Fyn, Lck, Tie ease, thyroid disorders, tumoral obstruction, fibrosing medi 2, GSK-3, Cdk2, Cdk4, MEK1, NEK-2, CHK2, CKle, Raf, astinitis, immunological and inflammatory diseases, hyper CHK1, Rsk2, FMS (CSF-IR), KDR, EphA2, EphA3, EphA8, proliferative diseases, renal and kidney diseases, bone FLT1, FLT4, HCK, PTK5, RET, SYK, DDR1, DDR2 and remodeling diseases, metabolic diseases, vascular diseases, PAR-1. In particular, the present disclosure provides com and chronic renal failure on dialysis. pounds, compositions, and methods of treating biological 0153. In one aspect, the present disclosure provides a conditions mediated by, or associated with, tyrosine kinases, method of treating pulmonary arterial hypertension (PAH) in including, but not limited to, Cdc2 kinase, AKT, c-Kit, a subject or a biological condition associated with PAH in a c-ABL, p60src, VEGFR3, PDGFRO, PDGFRB, PDGFR-Co., Subject by administering to the Subject a therapeutically PDGFR-B B, PDGFR-O?3, FGFR3, FLT-3, Fyn, Lck, Tie-2, effective amount of a compound of Structure 1, a tautomer of FMS (CSF-IR), KDR, EphA2, EphA3, Eph A8, FLT1, FLT4, the compound, a pharmaceutically acceptable salt of the com HCK, PTK5, RET, SYK, DDR1, and DDR2. In some pound, a pharmaceutically acceptable salt of the tautomer, or embodiments, the disease or condition mediated by, or asso a mixture thereof, wherein a compound of Structure 1 is ciated with, one or more kinases is mediated by a RTK, such described herein. In some embodiments, the disease or con as, e.g., PDGFR, PDGFR-CC, PDGFR-BB, PDGFR-CfB, or dition mediated by, or associated with, one or more kinases of c-Kit, or combinations thereof. the present disclosure is selected form the group consisting of 0152 The disease or condition mediated by, or associated PAH, primary PAH, idiopathic PAH, heritable PAH, refrac with, one or more kinases of the present disclosure, includes, tory PAH, drug-induced PAH, toxin-induced PAH, and PAH but is not limited to, PAH, primary PAH, idiopathic PAH, with secondary diseases. heritable PAH, refractory PAH, BMPR2, ALK1, endoglin associated with hereditary hemorrhagic telangiectasia, 0154 Pulmonary arterial hypertension (PAH) is a life endoglin not associated with hereditary hemorrhagic telang threatening disease characterized by a marked and Sustained iectasia, drug-induced PAH, and toxin-induced PAH, PAH elevation of pulmonary artery pressure. The disease results in associated with or secondary to one or more of systemic right ventricular (RV) failure and death. Current therapeutic Sclerosis, mixed connective tissue disease, cancer, refractory approaches for the treatment of chronic pulmonary arterial cancer, metastatic cancer, neoplasia, hypoplasia, hyperplasia, hypertension mainly provide symptomatic relief, as well as dysplasia, metaplasia, prosoplasia, desmoplasia, angiogenic Some improvement of prognosis. Although postulated for all disease, pulmonary function disorders, cardiovascular func treatments, evidence for direct anti-proliferative effects of tion disorders, HIV infection, hepatitis, portal hypertension, most approaches is missing. In addition, the use of most of the pulmonary hypertension, congenital heart disease, hypoxia, currently applied agents is hampered by either undesired side chronic hemolytic anemia, newborn persistent pulmonary effects or inconvenient drug administration routes. Pathologi hypertension, pulmonary veno-occlusive disease (PVOD), cal changes of hypertensive pulmonary arteries include pulmonary capillary hemangiomatosis (PCH), left heart dis endothelial injury, proliferation and hyper-contraction of vas ease pulmonary hypertension, Systolic dysfunction, diastolic cular smooth muscle cells (SMCs), and fibroblast prolifera dysfunction, valvular disease, lung disease, interstitial lung tion. PAH patient status, moreover, can be assessed in accor disease, pulmonary fibrosis, Schistosomiasis, chronic dance with the World Health Organization (WHO) obstructive pulmonary disease (COPD), sleep-disordered classification (modified after the NY Association Functional breathing, alveolar hypoVentilation disorders, chronic expo Classification) as known in the art. Sure to high altitude, developmental abnormalities, chronic 0.155. In some embodiments, the compounds of Structure thromboembolic pulmonary hypertension (CTEPH), pulmo 1 treat or prevent PAH in patients who failed prior therapy, nary hypertension with unclear multifactorial mechanisms, especially after receiving at least one prostanoid, endothelin hematologic disorders, myeloproliferative disorders, sple antagonist or PDE V inhibitor. In other embodiments, the nectomy, systemic disorders, sarcoidosis, pulmonary Langer compounds treat or prevent PAH in patients who are more hans cell histiocytosis, lymphangioleimoyomatosis, neurofi severely affected, in particular in patients with Class II to bromatosis, vasculitis, metabolic disorders, glycogen storage Class IV functional status, or more severely Class III or IV US 2015/0353527 A1 Dec. 10, 2015 22 functional status. In further embodiments, the compounds pared to the subject prior to the administering. Methods for treat or prevent PAH in patients who are harboring BMPR2 measuring lung function and blood pressure are known in the mutations. art. In one aspect, the present disclosure provides a method of 0156 The present disclosure provides methods of pre treating pulmonary arterial hypertension (PAH) in a Subject, venting or treating Subjects afflicted with idiopathic or pri comprising: modulating the phosphorylation-state (“PS) of mary pulmonary hypertension, familial hypertension, pulmo one or more downstream targets of platelet derived growth nary hypertension secondary to, but not limited to, connective factor receptor-alpha or platelet derived growth factor recep tissue disease, congenital heart defects (shunts), pulmonary tor-beta or both, wherein the downstream target is any sub fibrosis, portal hypertension, HIV infection, sickle cell dis strate phosphorylated as a result of the PDGFR-C. and/or the ease, drugs and toxins, e.g., anorexigens, cocaine, chronic PDGFR-B activation, by administering to the subject a com hypoxia, chronic pulmonary obstructive disease, sleep apnea, pound of Structure 1, a tautomer, enantiomer, isomer or Ste and Schistosomiasis, pulmonary hypertension associated reoisomer of the compound, a pharmaceutically acceptable with significant venous or capillary involvement (pulmonary salt of the compound, tautomer, enantiomer, isomer or stere Veno-occlusive disease, pulmonary capillary hemangiomato oisomer of the compound, or any mixtures thereof, wherein sis), secondary pulmonary hypertension that is out of propor the downstream target is selected from the group consisting of tion to the degree of left ventricular dysfunction, and/or per AKT, PDGFR, STAT3, ERK1 and ERK2, or any other down sistent pulmonary hypertension in newborn babies, especially stream target of the PDGFR-O. and/or the PDGFR-B, and in subjects that previously failed prior PAH therapy. wherein the compound of Structure 1 is described herein. 0157. In one aspect, the present disclosure provides a com Phosphorylation state profiles for proteins, kinases/receptors, pound of Structure 1, a tautomer of the compound, enanti omer, isomer or Stereoisomer of the compound, a pharmaceu can be ascertain using techniques known in the art, Such as, tically acceptable salt of the compound, tautomer, for example, Z-lyte kinase assays, Invitrogen Select Screen R, enantiomer, isomer or stereoisomer of the compound, or any and other kinases assays know in the art. mixtures thereof for treating one or more diseases associated (0162. In suitable embodiments, the modulation of the with hyperproliferation, neoplasia, hypoplasia, hyperplasia, kinase receptor activity is an inhibition of the kinase receptor dysplasia, metaplasia, prosoplasia, desmoplasia, angiogen activity. PDGFR, i.e., PDGFR-O, PDGFR-B, PDGFR-Co., esis, inflammation, pulmonary function, and cardiovascular PDGFR-BB, and PDGFR-O?3, and/or c-Kit are examples of function, where a compound of Structure 1 is described RTKs that are inhibited in some embodiments of the present herein. invention. In some embodiments, the inhibition is at least a 0158 Hyperproliferative, immunological and inflamma 0.001, 0.01, 0.1, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, tory, metabolic, and vascular diseases, are known in the art, 65, 70, 75, 80, 85,90, or 95% inhibition. In some embodi and such diseases, as described in U.S. Provisional Patent No. ments, the PSR modulation is a modulation of one or more of 61/751.217, which is hereby incorporated by reference in its AKT, STAT3, ERK1, ERK2, PDGF, and PDGFR i.e., entirety, are therapeutic targets for the compounds and agents PDGFR-CC, PDGFR-B B, and PDGFR-co.f3. In some described herein. embodiments, the modulation of PS is a decrease of phospho 0159. Another aspect of the present disclosure related to a rylated STAT3 to total STAT3 in the subject compared to the method of preventing or reducing elevated pulmonary pres PS in the subject prior to the administering. In some embodi Sure in a subject, by administering to the Subject a therapeu ments, the decrease is at least a 0.001, 0.01, 0.1, 1, 5, 10, 15, tically effective amount of a compound of Structure 1, a 20, 25, 30,35, 40, 45, 50,55,60, 65,70, 75,80, 85,90, or 95% tautomer of the compound, a pharmaceutically acceptable decrease. In some embodiments, the modulation of PS is a salt of the compound, a pharmaceutically acceptable salt of decrease of diphosphorylated ERK1 to total ERK1 in the the tautomer, or a mixture thereof, where a compound of subject compared to the PS in the subject prior to the admin Structure 1 is described herein. See, e.g., Summary. In some istering. In some embodiments, the decrease is at least a embodiments, the compounds of Structure 1 treat or prevent 0.001, 0.01, 0.1, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, a biological condition associated with PAH. Such as, e.g., 65, 70, 75, 80, 85, 90, or 95% decrease. In other embodi abnormal: right ventricular systolic pressure (RVSP); pulmo ments, the modulation of PS is a decrease of diphosphory nary pressure; cardiac output; right ventricular (RV) hyper lated ERK2 to total ERK2 in the subject compared to the PS trophy; and PA hypertrophy. in the Subject prior to the administering. In some embodi 0160. In some embodiments, the compounds of Structure ments, the decrease is at least a 0.001, 0.01, 0.1, 1, 10, 50, 60, 1 reduce pulmonary pressure associated with an increase in 70, 80, 85,90, or 95% decrease. one or more of right ventricular (RV) function, pulmonary (0163. In some embodiments, the modulation of PS is a artery (PA) systolic pressure, and/or cardiac output in the decrease of monophosphorylated ERK1 to total ERK1 in the Subject compared to the Subject prior to the administering. In subject compared to the PS in the subject prior to the admin Some embodiments, the reduction in pulmonary pressure is istering. In some embodiments, the decrease is at least a associated with a decrease in one or more of RV hypertrophy, 0.001, 0.01, 0.1, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, PA hypertrophy, RVSP sustained PA pressure, and the risk of 65, 70, 75, 80, 85, 90, or 95% decrease. In some embodi stroke in the subject compared to the subject prior to the ments, the modulation of PS is a decrease of phosphorylated administering. In some embodiments, the decrease is at least PDGFR to total PDGFR in the subject compared to the PS in a 5,10,15, 20, 25, 30,35, 40, 45,50, 55,60, 65,70, 75,80, 85, the Subject prior to the administering. In some embodiments, 90, or 95% decrease. In some embodiments, the decrease is at the decrease is at least a 0.001, 0.01, 0.1, 1, 5, 10, 15, 20, 25, least a 40% decrease. 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85,90, or 95% 0161. A reduction in pulmonary pressure, in some decrease. In some embodiments, the modulation of PS is a embodiments, is not associated with decreased lung function decrease of phosphorylated AKT to total AKT in the subject and/or increased systemic blood pressure in the Subject com compared to the PS in the subject prior to the administering. US 2015/0353527 A1 Dec. 10, 2015
In some embodiments, the decrease is at least a 0.001, 0.01, DMEM with 5% FBS and 4 mM Glutamax at 37° C., 5% 0.1, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65,70, 75, CO. HLFs were plated and grown to 70-80% confluence in 80, 85,90, or 95% decrease. 96 well plates then serum-starved for 48 hours. Cells were treated with drug (PK10453 or imatinib) at indicated concen EXAMPLES trations for 30 min then exposed to 10 ng/ml PDGFAA or BB 0164. The present invention is further illustrated by the for 7.5 min. Cells were fixed in 3.7% formaldehyde, washed following examples, which should not be construed as limit with 0.1% Triton X-100, and treated with Odyssey Blocking ing in any way. The following is a description of the materials Buffer for 90 min. Proteins were incubated overnight 1:100 and methods used throughout the examples, which illustrates diluted rabbit mAb to phosphorylated AKT (Ser 473 or Thr that RTK signaling pathways are activated in human disease 308) and 1:100 mouse mAb to total Akt-pan-4040D. Anti conditions, e.g., PAH and in animal models of the disease. bodies were detected using IRDye 680LT goat anti-mouse (0165 Materials. PK10453, (S)-N-(3-(1-((6-(4-hydroxy IgG and IRDye 800W goata-rabbit IgG conjugated antibod 3-methoxyphenyl)pyrazin-2yl)amino)ethyl)phenyl)-5-meth ies. After washing, the signal was quantified using an Odys ylnicotinamide, i.e., Structure 2, was synthesized by Organix, sey Infrared Imaging System (LI-COR). Phosphoprotein sig Inc. (Woburn, Mass.). Human PA smooth muscle cells and nal (800 nm) was normalized to total protein signal (700 nm) cell culture media were obtained from Cell Applications, Inc. acquired from each well and experimental duplicates on same PDGFBB, para-toluene sulfonic acid, ammonium hydroxide, plate were averaged and reported. and IR780 were obtained from Sigma Aldrich (St. Louis, 0169. Animals. Male Sprague Dawley rats (weight 320 Mo.). Imatinib mesylate was obtained from LC Laboratories 330 grams; Taconic Inc.) were used for this study. Animals (Woburn, MA). Human fetal lung fibroblasts (HLFs) were were housed in standard rat cages with a 12hlight/dark cycle, obtained from Cell Applications, Inc., San Diego. DMEM and standard rat chow and water were provided ad libitum. medium was obtained from Mediatech (Manassas, Va.). Animals were cared for and used in accordance with NIH PDGFAA, PDGFBB, and Glutamax were obtained from Life guidelines. All animal protocols were approved by the Bassett Technologies (Grand Island, N.Y.). Para-toluene sulfonic Medical Center and Pulmokine IACUC. acid, ammonium hydroxide, IR780 and monocrotaline (0170 Formulation and Aerosol Delivery. PK10453 (C2401 Lot 031M1921 V and LotSLBB7802V) were (Structure 2) was dissolved at a concentration of 20 mg/ml in obtained from Sigma Aldrich (St. Louis, Mo.). Anti-phospho 1M tosylic acid. Nebulization was performed with a PARI AKT(Ser 473), anti-phospho-AKT (Thr308), pan-AKT Nebulizer with an air pressure of 12.5 psi. The aerosol drop (CST2920 mouse maB, and CST2965 rabbit mAb), anti lets were neutralized by ammonia vapor that was passed into phospho-ERK 1/2, anti phospho-STAT3, and total STAT3 the aerosol air stream. The particles were then dried by flow antibodies were obtained from Cell Signaling Technologies, ing through an annular ring of silica bead cartridges prior to (Waltham, Mass.). Anti-total ERK1/2 antibody was obtained reaching the exposure chamber. The 6-port exposure chamber from Protein Simple (CA). AntiVon-Willebrand Factor, actin, was a nose-only exposure system custom designed and built phospho-PDGFRO. (Y754), and PDGFBB antibodies were by Powerscope Inc. (Minneapolis, Minn.). The vacuum flow obtained from AbCam (Cambridge, Mass.). Antibodies rate at each port was separately controlled by a flow meter. against PDGFAA (sc-128), PDGFR-alpha (sc-338), PDGFR The aerosol particle size was measured at the exit port of the beta (sc-432) and p-PDGFR-beta (Tyr 1021) (sc-12909) were drying column with an Anderson (Mark II) cascade impactor. obtained from Santa Cruz Biotechnology (CA). 680LT goat The mass median aerodynamic diameter (MMAD) was 2 um anti-mouse IgG, IRDye 800 W goat anti-rabbit IgG, and and the associated geometric standard deviation (GSD) was Odyssey blocking buffer were obtained from Licor (Lincoln, 1.6. Imatinib mesylate was dissolved in water at 20 mg/ml Nebr.). and delivered by a PARI nebulizer then dried by passage 0166 In vitro kinase assay. A Z-lyte kinase assay was through an annular ring of silica bead cartridges prior to performed to determine the inhibition of PDGFRalpha and inhalation. PDGFRbeta mediated phosphorylation by PK10453 (Struc 0171 Estimation of inhaled Dose. Filters exposed to ture 2). Ten point titration curves were modeled to calculate PK10453 (Structure 2) for either 4 or 8 min (n=6 each group) the ICs (Invitrogen Select Screen(R). via the Powerscope exposure chamber were placed in amber 0167 PASMC proliferation assay. Human pulmonary glass vials. Twelve milliliters of 1:3 (v/v) methanol:acetoni artery smooth muscle cells (PASMC) were obtained from trile were added to each vial containing a filter for approxi Cell Applications (San Diego, Calif.) and grown to 50% con mately 1 hr., with periodic mixing, followed by sonication for fluence in a 96 well format. The cells were switched to serum 60 seconds. An aliquot was then diluted 100-fold by adding free media 24 hours prior to stimulation with PDGFBB 50 10 uL of unknown filter extract to 990 uL of 1:3 (v/v) metha ng/ml and varying concentrations of PK10453 (Structure 2). nol:acetonitrile. Samples were vortex mixed for 30 seconds, After 24 hours of treatment, a Cycuant NF Cell proliferation and then a 100 uL diluted aliquot was combined with 100LL assay was performed (InvitrogenR), and the fluorescent sig of 172 ng/mL of a nonchemically related internal standard nal was measured with a Cytofluor Plate reader. Data is based (PK18855) in 1:1 methanol: water, vortex mixed and trans on an average of 8 replicates at each concentration. ferred to autosampler vials for LC-MS/MS analysis. Filter (0168. In cell Western (ICW). To compare the inhibitory extracts were compared against a calibration curve prepared profiles of PK10453 (Structure 2) and imatinib for PDGFBB in 100% methanol (Pharmoptima R, Inc.). The aerosol con and PDGFAA stimulated AKT phosphorylation, ICWs were centration of PK10453 (Structure 2) in lug/liter of air was performed, with modifications, according to the method of calculated based on the average total mg of PK10453 (Struc Chen et al., “A cell-based immunocytochemical assay for ture 2) on the filters for the 4 and 8 min exposure times, and monitoring kinase signaling pathways and drug efficacy.” the flow rate past each filter (0.8L/min) The inhaled dose was Analytical biochemistry; Vol.338:136-42 (2005). HLFs were calculated with the average concentration of PK10453/cm maintained in Subculture at no more than 6 passages in filter paper (average of 4 and 8 min exposures), the average US 2015/0353527 A1 Dec. 10, 2015 24 minventilation measured by plethysmography (0.15 L/min), exposures of either the vehicle control or PK10453 (Structure and an estimated deposition fraction of 0.1. The imatinib 8 2). Sampling of PA pressure was performed 5 min before each min dose was based on gravimetric analysis. morning dose in ambulatory animals in room air (estimated 0172 Imaging. The spatial distribution of inhaled atmospheric pressure 716 mm Hg based on elevation of ani PK10453 (Structure 2) in the lung was evaluated by fluores mal facility). In protocol 4 (imatinib vs. vehicle), the animals cent imaging. A near IR fluorescent tracer, IR-780, was added received DSI PAC40 transmitters followed by monocrotaline to the drug solution in the nebulizer to ensure dried aerosol 50 mg/kg IP (Lot SLBB7802V). A lower dose of MCT was particles contained both the drug and IR tracer. After a two used for this study, because attempts to use 60 mg/kg of this min exposure, animals were placed under general anesthesia lot of MCT resulted in the need for early euthanasia in a high underwent intubation via tracheostomy, and the lungs were proportion of animals due to weight loss and tachypnea. Two excised. OCT/PBS was infused via the pulmonary artery, the weeks after MCT IP injection, vehicle (mesylate 3 mg/ml) or lung insufflated with air, and the lungs frozen in the vapor imatinib mesylate 20 mg/ml in nebulizer solution) was phase of liquid nitrogen. Serial approximate 2mm sections of administered for 8 min exposures three times a day for 9 days. lung were imaged on a Licor Odyssey Imager. Telemetry data was obtained for 10 min daily before each (0173 Pharmacokinetic studies. PK10453 (Structure 2) morning dose for this protocol. was administered intravenously or by inhalation to animals, 0177. Measurement of PV loops. In a separate cohort of which were then euthanized at time 0, 10, 20, and 60 min (n=3 animals, the MCT+PN model was developed as described each time point). Blood samples were taken by cardiac punc above, and PK10453 (Structure 2) was then administered for ture, and the lungs excised. The lungs were homogenized and 4 or 8 min three times a day to the drug treated group. The PK10453 (Structure 2) extracted with a 1:3 mixture of aceto vehicle control group underwent 4 min exposures three times nitrile:methanol. Similarly, plasma was extracted with a 1:3 a day. Pressure Volume (PV) loops were obtained with an mixture of acetonitrile:methanol. Drug was assayed by LC admittance system (SciSense, Inc.) after 14 days of treatment, MS/MS (PharmCptima Inc., Portage Mich.). First order while rats were under general anesthesia with isoflurane and exponential curves were fit to the data with Excel. AUC was 100% FiO. Also, or in the alternative, RV pressures were determined with the trapezoidal method of integration. obtained in each group after 14 days of treatment. In a Subset (0174) Efficacy study in the rat MCT model PK10453 of each group, pressure Volume (PV) loops were obtained (Structure 2) dose response study in the rat MCT Model. Male with an admittance system (high fidelity catheter FTE 1918B, Sprague Dawley rats received MCT 60 mg/kg IPMCT, and Scisense, Inc.) after 14 days of treatment. After induction of after 3 weeks, PK10453 (Structure 2) or vehicle control were general anesthesia and intubation via tracheostomy, the rats administered by inhalation. Four groups were studied: were placed on a pressure controlled ventilator (TOP vehicle control (4 min exposure) and three treatment groups OVENT). General anesthesia consisted of isoflurane and of PK10453 (Structure 2) with exposure times 2 min (D2), 4 100% FiO with peak inspiratory pressure set at 18 cm, and min (D4), or 8 min (D8) three times a day. These regimens PEEP 5 cm H2O. A left thoracotomy was performed with were administered for two weeks. The vehicle consisted of admittance catheter in the RV via the RV outflow tract. aerosolized 1M tosylic acid neutralized with ammonia vapor (0178 Systemic blood pressure study. The effect of as described above. The pH of a solution prepared by dissolv PK10453 (Structure 2) on systemic BP was studied in ambu ing captured aerosol particles in water was measured for latory MCT treated rats with DSI PAC40 transmitters every dose and was consistently in the range of 5.5-6.0. At the implanted in the descending aorta. Three weeks after admin end of the study, the RV systolic pressure was measured, and istration of MCT 60 mg/kg IP animals inhaled PK10453 the heart chambers dissected and weighed. (Structure 2) or vehicle 3x/d with 4 min exposure for 7 days. (0175 Efficacy study in the rat MCT model PK10453 Blood pressure was recorded before each morning dose. (Structure 2) vs. imatinib in the rat MCT model. Male Spra 0179 Plethysmography. Plethysmography was performed gue Dawley rats were given MCT 60 mg/kg IP. Three weeks with an EMKA dual chamber plethysmograph and IOX soft later vehicle (1M tosylic acid), PK10453 (Structure 2 at 20 ware. Parameters measured included breathing frequency, mg/ml free base in 1 Mtosylic acid), or imatinib mesylate (20 tidal Volume, minute ventilation, peak inspiratory and expi mg/ml in nebulizer Solution) were administered to designated ratory flow, and airway resistance (SRaw). Animals were groups for 8 min inhalation exposures, three time a day, for acclimatized to the plethysmograph for three days prior to two weeks. At the end of the study RVSP pressure was mea first data acquisition. Measurements were made prior to the Sured; lung and heart fixed in formalin. For measurement of first dose of drug and at the end of the study. RVSP animals were sedated with isoflurane, intubated via a 0180. Histology and morphometric analysis. At the end of tracheostomy, and ventilated with a TOPOVENT pressure the study, the heart and lungs were removed from ventilated regulated ventilator (peak inspiratory pressure 18 cm H2O, animals under general anesthesia. Heparinized saline was PEEP 5 cm). After sternotomy, a Scisense high fidelity cath infused under pressure through the main pulmonary artery. eter inserted via the RV apex. The right upper lobe was immediately tied off and placed in (0176 Efficacy study in the rat MCT+PN model. Pneu liquid nitrogen for Western blot and NanoPro 100 assay monectomy and implantation of a TRM53P telemetry moni analysis. The heart was removed, and the RV free wall, inter tor in the pulmonary artery (Telemetry Research, New ventricular septum and LV free wall dissected and weighed. Zealand and ADInstruments, Colorado) was carried out in Buffered formalin (10%) was infused under pressure both rats. Two weeks after MCT, PK10453 (Structure 2) was through the pulmonary artery and the trachea. Morphometric administered three times daily for 1 week. Dosing was begun analysis was performed on H&E stained formalin fixed tissue 2 weeks after MCT rather than 3 weeks, because in this more sectioned at 8 Lum. The media area and lumen area of pulmo aggressive model the animals developed PAH more quickly nary arterioles were measured with Image J Software by a and developed distress sooner than in MCT only treated ani technician blinded to treatment group. Measurements were mals (data not shown). The two groups underwent 4 min made on 20 pulmonary arterioles persection. The ratio of the US 2015/0353527 A1 Dec. 10, 2015
lumen area to the total media area was determined This ratio 001). The ICs concentrations of PK10453 and imatinib for normalizes the variation in total pulmonary arteriole area. In PDGFAA stimulated phosphorylation of AKT were not sig addition, occlusive analysis was performed in the monocro nificantly different. taline plus pneumonectomy study (specifically efficacy study 5)according to the method of Homma et al., “Involvement of 0185. Estimated inhaled dose - PK10453 (Structure 2) and RhoA/Rho kinase signaling in protection against monocrota imatinib. The average concentration of PK10453 was line-induced pulmonary hypertension in pneumonectomized 62.4+3.3 ug/cm filter paper for the 4 min exposure, and rats by dehydroepiandrosterone.” Am J Physiol Lung Cell 137+7.0 g/cm for the 8 min exposure, which resulted in an Mol Physiol. Vol. 295:L71-8 (2008). Briefly, pre-capillary aerosol concentration of 91.65ug/L airfor the 4 min exposure arterioles were assigned grade 0 for no evidence of neointimal and 100.6 g/L air for the 8 min exposure. The aerosol con lesions, grade 1 for less than 50% luminal occlusion, and centration of imatinib based on gravimetric analysis was 167 grade 2 for greater than 50% occlusion. Masson Trichrome ug/L. The average inhaled dose (8 min), assuming a deposi stains were performed on lung sections from the MCT+PN tion fraction of 0.1 and rat weight 300 g, was approximately model. 20 g/kg for PK10453, and 40 ug/kg for imatinib, as shown in Table 1. The estimated inhaled dose was calculated from the 0181 NanoPro Immunoassay. Relative differences in measured concentration of PK10453 (Structure 2) and gravi phosphorylated ERK 1/2 and STAT isoforms were measured metric analysis of imatinib in the aerosol, the measured with a NanoPro 100.R immunoassay system (Protein Simple/ minute ventilation (MV), the estimated deposition fraction of Cell Biosciences, CA). See Fanet al., “Nanofluidic proteomic 0.1, and rat weight 300 g. TABLE 1. Aerosol Cone MV*Exposure Total Deposition API Lig/L Exposure Min MV L/min time fraction
PK10453 96.13 8.00 O.15 1.20 O.10 Imatinib 167.4 8.00 O.15 1.20 O.10
Lung Deposition Total Total Lung Lung deposited API fraction Inhaled Ig Deposited jug Deposited Ig |lgkg
PK10453 O.60 115.36 11.54 6.92 23.07 Imatinib O.60 200.88 20.09 12.05 40.18 assay for serial analysis of oncoprotein activation in clinical 0186 Lung distribution and pharmacokinetics of inhaled specimens.” Nat Med 15:566-571 (2009). PK10453 (Structure 2). Fluorescent images of the lung sec 0182 Immunohistochemistry. Antigen retrieval was per tions following inhalation of PK10453 with IR780 tracer are formed with citrate buffer (pH 6.0) or Tris-EDTA buffer (pH shown in FIG.3, where the flurorescence intensity is shown to 9.0). Immunohistochemistry was performed for the following be well distributed throughout the lungs. The network of targets: CD20 (a B cell marker), CD3 (a T cell marker), von darker lines arises from the connective tissue and therefore Willebrand Factor (vWF), total STAT3, phosphoSTAT3 does not represent the airways affected by the disease. The (Tyr705), total PDGFR-alpha, total PDGFR-beta, and phos spatial distribution of imatinib was similar (data not shown). phoPDGFR-beta. Competing peptides were available for 0187. For the pharmacokinetic study, the concentration of PDGFR-alpha and phospho-PDGFR-beta. Signal detection PK10453 (Structure 2) in lung when administered by inhala was performed with an EXPOSE HRP/DAB kit (AbcamR). tion was compared to the concentration achieved with IV 0183 Statistical Analysis. Data are presented as administration. As described in Moren Aerosols in medicine meant SEM unless otherwise noted. The General Linear : principles, diagnosis, and therapy.” Amsterdam; New York: Model with the Bonferroni correction for multiple group Elsevier. XX, 429 (1993) and Phalen et al., “Inhalation expo comparisons was used (SPSS 14.0). Significance was set at sure methodology.” Environ Health Perspect 56:23-34 the p=0.05 level. (1984)), it is possible to estimate the pharmacokinetic advan tage of inhalation relative to intravenous administration, R. Example 1 by comparing the AUC of a plot of the drug concentration as a function of time following respiratory and IV administra Characterization of PK10453 (Structure 2) tion: Ri(AUCAUC)respiratory (AUC, 0184 An in vitro kinase assay demonstrated the ICs for AUC)IVpiasna PK10453 at ATP K was 35 nM for PDGFR-O, and 10.1 nM for the PDGFR-3. For imatinib the ICs at ATPK was 71 nM 0188 The pharmacokinetic data was modeled to a first for PDGFR-O and 607 nM for PDGFR-beta. See FIG.1. The order exponential curve, and the AUC calculated from the ICs of PK10453 for PDGFBB stimulated AKT phosphory curves (see Table 2). FIG. 4 shows the drug level in lung and lation at Ser473 was 0.13 uM compared to 1.8 LM for ima plasma as a function of time following inhalation or IV tinib (P<0.01), as shown in the ICW illustrated in FIG. 2. The administration of PK10453 (Structure 2). The data indicate a ICs of PK10453 for PDGFBB stimulated AKT phosphory 45 fold advantage of inhaled compared to IV administered lation at Thr308 was 0.43 uM vs. 3.25uM for imatinib (p<0. PK10453 (R-44.6). US 2015/0353527 A1 Dec. 10, 2015 26
TABLE 2 days of treatment, the PA systolic pressure was 67.4+ 11.4 mm Hg in the vehicle group and was significantly lower at 47.2-3 Y = AEXP(-bX) A (ngig lung) b (min-1) R2 AUC 0 mm Hg in the PK10453 group (p=0.03). On treatment day Lung (INH) 2498 O.O3 O.89 1OO1.82 9, the PA systolic pressure in the vehicle group was 92.8+9.1 Plasma (INH) 132.7 O.O7 O.93 65.47 mm Hg, but significantly lower at 50.5-7 mm Hg in the Lung (IV) 440 O.O6 O.96 211.89 PK10453 group (p=0.03). For the imatinib telemetry study Plasma (IV) 1260 O.O7 O.92 617.25 (study 4), at day 1, the PA systolic pressure in the vehicle Rd group was 51.4-8.9 mm Hg, and in the imatinib group 4458 41.5-3.5 mm Hg. At treatment day 9 the PA systolic pressure in the vehicle group was 80.4-14.2 mm Hg, and in the ima tinib group was 75.1+7 mm Hg (p=NS). See FIG. 6. Example 2 (0193 Measurement of RV pressure and PV loops in the MCT+PN model; PK10453 (Structure 2) dose response MCT Model Efficacy study. In a separate cohort of animals, the MCT+PN model was developed as described. RV pressure was obtained after (0189 RVSP values are shown in FIG. 5A. In the vehicle 14 days of vehicle exposure, and PK10453 treatment with 4 group (n=6), RVSP was 80.4+2.6 mm Hg. For the treatment min (D4) and 8 min exposures (D8) three times a day. In the groups, D2 (n=6), 51.4+6.5; D4 (n=6), 44.4+3.8; and D8 vehicle group (n=9), RV systolic pressure was 75.7+7.1 mm (n=5), 37.1+4.5 mm Hg (p<0.001). Normal control group Hg, in the D4 group (n=10) RV systolic pressure was RVSP was 28.5+2.6 mm Hg (n=3). In the D4 group, there was 40.4+2.7 mm Hg, and in the D8 MCT+PN group RV systolic a 44% reduction in RVSP, and in the D8 group, there was a pressure was 43+3.0 mm Hg (p<0.001 V vs. D4 and V vs. D8; 54% reduction in RVSP compared to the vehicle treated FIG. 7A). PV loops were obtained in a subset of animals from group. There was also a significant reduction in the degree of each group (Vehicle n=3; D4 n=5, D8 n=4). RV hypertrophy as measured by the ratio (RV+IVS)/LV weight. See FIG. 5B. The data are represented by this ratio Example 4 because the septum is shared by the RV and LV. However, use of the RV/(IVS+LV) ratio also showed similar results. MCT+PNMCT+PN Model Efficacy 0190. Moreover, there were 6 animals in the vehicle group but accurate RV end systolic pressures were not obtained due (0194 PV loop study. The RV end systolic pressure (ESP) to bleeding in 2 animals. Therefore RV systolic pressure is was lower and the RV ejection fraction (EF) was higher in based on n=4 in the vehicle group and was 57.9+7.6 mm Hg. both the D4 and D8 treatment groups compared to vehicle In the PK10453 (Structure 2) group (n=12) RV end systolic control. Cardiac output in the D8 group was increased com pressure was 36.32 6 mm Hg, and in the imatinib group pared to the Vehicle group. See Table 3. The study animals (n-6) was 31.8+1.8 mm Hg (p=0.001 Vehicle vs. PK10453; underwent left pneumonectomy followed 7 days later by p=0.002 Vehicle vs. Imatinib, FIG.5C). End systolic volume MCT 60 mg/kg IP. Two weeks after MCT administration, was greater in the vehicle group (158+12.6 ul) vs. PK10453 PK10453 (Structure 2) or vehicle were given by inhalation (99.5+10 ul) and imatinib (81+4.3 ul) (p=0.05 vehicle vs. three times a day for two weeks. PV loops were acquired at PK10453; p=0.014 vehicle vs. imatinib; p=NS PK10453 vs. the end of this period. With respect to Table 3: V-vehicle: imatinib). There were no significant differences between the D4=4 min inhalation PK10453; D8–8 min inhalation groups for the following parameters: end diastolic Volume, PK10453: n-4 each group: *p-0.001: **p<0.01; Sp-0.05 vs. ejection fraction, cardiac output, stroke work. The lumen to V. See Table 3 below. media ratio was improved by both PK10453 and imatinib compared to vehicle in the MCT model (Vehicle (V. n=4): TABLE 3 0.55+0.1; PK10453 (D8, n=12): 0.94+0.08: Imatinib (18, HR ESP EDP ESV n=5): 0.99+0.07: p30.01 D8 vs. V, p<0.0518 vs. V. FIG.5D). Group Sel (bpm) (mm Hg) (mm Hg) (II)
Example 3 V Mean 290 83.21 10.31 484.17 SEM 25 3.49 1.24 148.32 D4 Mean 288 43.20% 2.62s 144.14 Efficacy Studies in the Rat MCT+PN Model SEM 21 6.08 O.30 25.89 0191 Telemetry studies. The results of the telemetry study D8 Mean 315 38.44 4.87 155.40 in the rat MCT+PN model are described. At day 0 prior to start SEM 41 1.43 186 22.69 of treatment, the PA systolic pressure in the vehicle groups ESV SV CO was 41.0+11.7 mm Hg, and in the PK10453 (Structure 2) Group Sel (II) (l) ml/min EF SW group, was 43.1+3.5 mm Hg (p=NS). After five days of treat V Mean 621.32 137.15 39.03 25.43 101.23 ment, the PA systolic pressure was 69.4+12 9 mm Hg in the SEM 139.49 1419 O.62 8.36 2698 vehicle group and was significantly lower at 47.3+3.0 mm Hg D4 Mean 408.95 264.81 77.59 654** 98.18 in the PK10453 group (p<0.01). On treatment day 8, the PA SEM 34.94 1266 2.59 3.47 769 systolic pressure in the vehicle group was 83.5-8.5, but sig D8 Mean 488.68 333.28** 105.1* * 67.1 * * S481 nificantly lower at 47.3+4.9 mm Hg in the PK10453 group SEM S2.OO 49.81 15.51 4.59 1829 (p<0.001). (0192. In a separate PK10453 (Structure 2) telemetry (0195 Effect of PK10453 (Structure 2) on RV hypertrophy. study, at day 1 prior to start of treatment, the PA systolic Treatment with PK10453 resulted in a significant decrease in pressure in the vehicle group was 47.4+10.2 mm Hg, and in RV hypertrophy in the rat MCT+PNMCT+PN model. See the PK10453 group, was 43.1+3.5 mm Hg (p=NS). After five FIG.7B. The (RV+IVS)/LV ratio in the vehicle group (n=11) US 2015/0353527 A1 Dec. 10, 2015 27 was 0.88+0.05, in the PK10453 D4 group (n=13) was 0.62+0. in precapillary pulmonary arterioles. Minimal signal for 04, and in the PK10453 D8 group (n=7) was 0.68+0.05 (p<0. pPDGFR-B or alpha was detected in the medial layers of the 001 D4 vs. V: p=0.012 D8 vs. V). pre-capillary pulmonary arterioles. See FIGS. 12A-F. In 0196. Analysis of pulmonary arteriole histology and mor larger (>50 um) vessels, signal for pPDGFR-oc was present in phology. The lumen area to media area ratio (L/M) was sig medial VSM cells. In contrast, pPDGFRB medial layer signal nificantly higher in the PK10453 (Structure 2) treated D8 was low. See FIGS. 13 A-D. group compared to the D4 or vehicle groups: D8 (n=5) L/M 0.72+0.05, D4 (n-6) L/M 0.33+0.06, and the vehicle control Example 6 V (n=6): 0.26+0.04 (p<0.0001 D8 vs. V or D8 vs. D4). See FIG. 7C. Occlusion analysis was performed on the same NanoPro(R) Immunoassays and Western Blots animal samples used for the lumen/media ratio measure ments. The occlusion analysis demonstrated a significant 0200 Nanopro(R) Immunoassays for p AKT/AKT are reduction in Grade 2 occlusion lesions in the PK10453 D8 shown in FIG. 14 and pSTAT3/STAT3 are shown in FIG. 15. treatment group (V (n-6) 41.5+7.1%, D4 (n-6) 28.5+4.2%; There was a significant reduction in the pSTAT3/STAT3 ratio D8 11.4+4.1%; p-0.01 D8 vs. V; see FIG.7D. FIG.8A shows in both the D4 and D8 groups compared to vehicle. FIG. 16 an H&E stain of an occlusive (Grade 2) lesion in a vehicle shows the effect of inhaled PK10453 (Structure 2) on treated animal (MCT+PN model); comparison is made to a ppERK1/ERK1, pERK1/ERK1, ppERK2/ERK2 and Grade 0 vessel from a PK10453 (D8) treated animal. See FIG. pERK2/ERK2 in lung homogenates. There were significant 8B. An example of a Grade 2 lesion stained for phospho reductions in ppERK1/ERK1 and pERK1/ERK1 in the D4 PDGFRbeta is shown in FIG. 8C with comparison to a Grade and D8 groups, respectively, compared to vehicle. 0 lesion from a PK10453 (D8) treated animal (MCT+PN model) in FIG.8D. Staining for phosphoPDGFRbeta showed Example 7 intense signal in a cobblestone pattern in Grade 2 lesions. 0.197 Futher examples of pulmonary arteriole hypertro PDGFAA Stimulates PDGFR-O, whereas PDGFBB phy and intraluminal cellular proliferative lesions are shown Binds & Activates PDGFR-B. as described, while the quantitative analysis is represented in 0201 FIG. 17 shows the effect of imatinib, PK10453 FIG.9. The lumen area to media area ratio (L/M) was signifi (Structure 2), and PK10571 (Structure 2a) on PDGFAA vs. cantly higher in the PK10453 treated groups compared to PDGFBB stimulated phosphorylation of ERK1 and ERK2 in vehicle, where the higher dose, D8 (n=4) L/M 1.17+0.07, the human fetal lung fibroblasts. The ratio diphosphorylated lower dose, D4 L/M 0.75+0.14, and the vehicle control V ERK1 to total ERK1 (ppERK1/ERK1) was increased with (n=6): 0.36+0.09 (p=0.032 D4 vs. V: p=0.00014 D8 vs. V: PDGFAA or PDGFBB stimulation, and significantly p=0.028 D8 vs. D4). The endothelial cell marker, VWF, decreased at 1 uM and 10 uMy concentration of imatinib, showed signal predominantly within the pulmonary arteri PK10453, and PK10571. The ratio diphosphorylated ERK2 oles. The tyrosine705 phosphoSTAT3 antibody showed to total ERK2 (ppERK2/ERK2) was increased with PDG localization of pSTAT3 to nuclei of endothelial cells and FAA or PDGFBB stimulation (10 ng/ml), and significantly perivascular cells. See FIGS. 10A; and FIG. 10B (with decreased at 1 uM and 10 uM concentration of imatinib, PK10453 treatment). PK10453, and PK10571. After PDGF BB stimulation, the 0198 Trichrome and Immunohistochemistry for alpha ratio of diphosphorylated ERK1 to total ERK1 (ppERK1/ SMC actin, and v Wif. The endothelial cell marker, v WF, ERK1) and diphosphorylated ERK2 to total ERK2 (ppERK2/ showed signal predominantly within the pulmonary arteri ERK2) was more effectively decreased at 1 uM PK10453, oles. And immuno-histochemistry for vascular SMCs (aSMC and PK10571 compared to imatinib. Thus, PK10453 and actin), endothelial cell markers (VWF) and trichrome stains of PK10571 are more potent inhibitors of PDGF BB stimulated pulmonary arterioles in the rat MCT+PN was performed to ERK1 and ERK2 phosphorylation compared to imatinib. further characterize Grade 0, 1, and 2 lesions. Grade Olesions 0202 In particular, and with reference to FIG. 17A-D as were characterized by early neointimal (intraluminal) prolif noted above, PDGFAA and PDGFBB (10 ng/ml) stimulation eration of endothelial cells (ECs) with preservation of vascu of human fetal lung fibroblasts increased ppERK1/ERK1 and lar SMCs in the media; Grade 1-2 lesions, by neointimal ppERK2/ERK compared to serum free media only controls (intraluminal) proliferation/invasion of mixed myofibroblast (SF). Imatinib, PK10453 (Structure 2), and PK10571 (Struc like cells (MFs) and ECs with partial loss of vascular smooth ture 2a) were equally effective at 1 uM in decreasing PDGF muscle cells in the medial layer; and advanced Grade 2 AA stimulated ppERK1 and ppERK2 formation (FIG. 17A lesions, by extensive MF/EC intraluminal proliferation with and C). However, PK10453 and PK10571 were more effec complete loss of VSM cells in the medial layer and fibrotic tive at 1 uM and 10 uM in decreasing PDGF BB stimulated replacement of the media. See FIGS. 11 A-I. ppERK1 and ppERK2 (FIG. 17B and D). These data demon Strate that PK10453 and PK10571 are more effective in Example 5 blocking signal transduction through the PDGF receptor beta Immunohistochemistry for PDGF Signaling compared to imatinib. Data shown are meant-SEM. The dif ferential effect of PK10453 and PK10571 was more promi 0199. In pre-capillary pulmonary arterioles signaling nent in blocking ERK1 vs. ERK2 phosphorylation. At 1 uM through the PDGFR-B pathway was dominant. Signal for imatinib had no effect on inhibition of ppERK1 formation PDGFAA ligand and PDGFR-u were present but qualita whereas PK10453 and PK10571 at 1 uM were effective in tively lower than signal for PDGFBB and PDGFR-B. Phos decreasing PDGFBB stimulated ppERK1 formation. phorylated PDGFR-B (ppDGFR-B) had a cobblestone PK10453=Structure 2: PK10571=Structure 2a. Platelet appearance in neointimal cells and in perivascular cells and derived growth factor receptor alpha=PDGFR was stronger than signal for phospho-PDGFR-C. (PDGFR-C) alpha=PDGFR-C=PDGF receptor alpha=PDGF alpha recep US 2015/0353527 A1 Dec. 10, 2015 28 tor. Platelet derived growth factor receptor beta=PDGFR TABLE 4-continued beta=PDGFR-13=PDGF receptor beta=PDGF beta receptor. (day 15) Example 8 Drug Group PIF PEF TV MV f SRaw
V mean 4.97 5.86 O.S2 107.59 21443 38.93 PK10453 (Structure 2), PK10467 (Structure 3), (n = 6) sem 0.39 0.44 O.O7 9.58 13.83 6.53 PK10468 (Structure 4), PK10569 (Structure 5) and D4 mean 7.82 9.338 0.85 176.12s 217.12 33.09 PK10571 (Structure 2a) Possess Lower ICs (n = 5) sem 0.34 0.67 O.12 14.53 18.64 4.8O Concentrations Compared to Imatinib for Inhibiting D8 mean 6.06 6.64 O.63 128.49 232.11 49.26 PDGFBB Stimulated AKT Phosphorylation in (n = 5) sem 0.84 O.99 O16 19.47 30.71 7.11 Fibroblasts Abbreviations: PDF; peak inspiratory flow; PEF: peak expiratory flow; 0203 Fetal human lung fibroblasts grown in cell culture TV; tidal volume; are used as a model of fibroblast proliferation that occurs in MV minute ventilation; pulmonary arterial hypertension, pulmonary fibrosis, and fbreathing frequency (breaths perminute); SRaw: airway resistance related disorders. See FIG. 18A-D. These data highlight that *p < 0.01 D4 vs. V: PK10453, PK10467, PK10468, PK10569, and PK10571 are Sp = 0.02 D4 vs. V. more potent inhibitors of signal transduction mediated through the PDGFbeta receptor compared to imatinib. These data show the importance of effective inhibition of PDGF Example 10 beta receptor signaling in addition to PDGF alpha receptor signaling as a treatment for pulmonary arterial hypertension, Discussion and Applied Embodiments pulmonary fibrosis, and related disorders which can be 0205 The PDGF signaling pathway has been found to be achieved with PK10453, PK10467, PK10468, PK10569, and activated in human pulmonary arterial hypertension (PAH) PK10571. As used above and throughout the application, the and in animal models of the disease. This study tested the PK compounds and structure designations are used inter hypothesis that a novel, non-selective inhaled PDGF receptor changeably, as follows: PK10453=Structure 2: inhibitor, PK10453 (Structure 2), would decrease pulmonary PK10571=Structure 2a: PK10467=Structure 3: hypertension both in the rat monocrotaline (MCT) model and PK10468=Structure 4; and PK10569=Structure 5. See FIG. the rat MCT plus pneumonectomy (+PN) model of PAH. 18A-D. PK10453 delivered by inhalation, for four (D4) and eight (D8) min exposures three times a day for two weeks, Example 9 decreased right ventricular systolic pressure (RVSP) in both the rat MCT and rat MCT+PN models: vehicle MCT group (n-6) RVSP was 80.4+2.6 mm Hg; in the D4 MCT group Body Weights, Systemic BP, and Plethysmography (n=6), 44.4+5.8 mm Hg; and in the D8 MCT group (n=5), Studies 37.1+4.5 mm Hg (p<0.001 vs. vehicle); in the vehicle MCT+ PN group (n=9) RVSP was 75.7+7.1 mm Hg; in the D4 0204 Compared to vehicle, there was a trend to a slower MCT+PN group (n=10), 40.4+2.7 mm Hg, and in the D8 rate of decline in body weight in the treated vs. vehicle MCT+PN group (n=8), 43.0+3.0 mm Hg (p<0.001). In the rat groups. See FIG. 19. On day seven of treatment, systolic BP MCT+PN model, continuous telemetry monitoring of pulmo was 111+21 mmHg in the MCT vehicle group (n-3) com nary artery pressures also demonstrated that PK10453 pre pared to 131+10 mmHg in the MCTPK10453 group (n=3), as vented the progression of PAH. Imatinib given by inhalation shown in FIG. 20. Two-chamber plethysmography was mea was equally effective in the MCT model, but was not effective sured at day 1 and day 15 of PK10453/vehicle administration in the MCT+PN model. in the rat MCT+PNMCT+PN model. The results are shown in 0206. Immunohistochemistry demonstrated increased Table 4. Treatment with PK10453 was associated with a activation of the PDGF13 receptor compared to the PDGFix slower decline in minute ventilation (MV), and a significant receptor in neointimal and perivascular lesions found in the improvement in peak inspiratory flow (PIF) and peak expira MCT+PN model. It was shown that imatinib is selective for tory flow (PEF) in the 4 min exposure group (D4) compared the PDGFix receptor whereas PK10453 has a lower ICs for to vehicle. inhibition of kinase activity of both the PDGFa and PDGFI3 receptors compared to imatinib. PK10453 decreased the ratio TABLE 4 of phosphorylated AKT (Ser473) to total AKT phosphory (day 1) lated STAT3 (Y705) to total STAT3, the ratio of diphospho rylated ERK1 to total ERK1 and the ratio of monophospho Drug TV MV rylated ERK1 to total ERK1 in lung extracts from MCT+PN Group PIF PEF (ml) (ml/min) f SRaw animals. In short, PK10453, when delivered by inhalation, V mean 8.81 9.68 O.86 19366 244.79 40.37 significantly decreased the progression of PAH in the rat (n = 6) Sel 0.79 O.98 O.14 20.66 28.02 4.11 MCT and MCT-PN models. Non-selective inhibition of both D4 mean 9.82 11.04 1.OO 223.24 224.68 39.73 PDGFC. and PDGFB receptors therefore has a therapeutic (n = 5) Sel O.70 O.S6 O.O7 11.99 9.87 3.33 D8 mean 8.54 9.43 O.74 174.68 259.13 36.01 advantage over the selective inhibition of PDGFRC. at least in (n = 5) Sel 0.72 O.O1 O.15 22.32 26.42 3.82 PAH and related diseases. 0207. Accordingly, and for the first time, it has been shown that a novel, non-selective PDGF receptor inhibitor, PK10453 US 2015/0353527 A1 Dec. 10, 2015 29
(Structure 2), when administered by inhalation, decreased the the neointimal lesions in the rat MCT+PN model have high severity of PAH in two animal models of the disease: the rat levels of phospho PDGFRB, with less pPDGFRC. These find MCT, and the rat MCT+PN model. As such, because ings explain why non-selective inhibition of both PDGFRB PK10453 is highly potent against both the PDGFRC. and and PDGFRC. provided a therapeutic advantage over the PDGFRB receptors, while imatinib is selective for the PDG Selective inhibition of PDGFRO. FRO. receptor, PK10453 possesses surprisingly superior effi 0211. The present data are consistent with Panzhinskiyet cacy. Both PK10453 and imatinib were effective in the rat al., “Hypoxia induces unique proliferative response inadven MCT model, but only PK10453 decreased pulmonary hyper titial fibroblasts by activating PDGFbeta receptor-JNK1 sig tension in the rat MCT+PN model when administered by naling.” Cardiovasc Res; 95:356-65 (2012), for the neonatal inhalation. One reason for this differential effect may be due calve model of high altitude induced pulmonary hyperten to hyper-activation of signaling through the PDGFRB recep sion. In that model extensive perivascular proliferation of tor in precapillary pulmonary arteriole neointimal lesions adventitial fibroblasts was demonstrated along with activa compared to the PDGFRC. receptor in the rat MCT+PN tion of pPDGFRB. These lesions are similar to the pattern model. observed in the rat MCT+PN model for the present studies. 0208 Accordingly, the present data demonstrates that a These findings are also consistent with those reported for novel, non-selective, PDGF receptor inhibitor PK10453 human PAH. Perros et al., “Platelet-derived growth factor (Structure 2) when delivered by inhalation prevented the pro expression and function in idiopathic pulmonary arterial gression of PAH in both the rat MCT and the rat MCT+PN hypertension.” Am J Respir Crit Care Med; 178:81-8 (2008), models. Of note, this is the first study to report efficacy of describing the distribution of PDGFA, PDGFB, PDGFRC, PDGF receptor inhibition in the rat MCT+PN model. A sus PDGFRB and pPDGFRB in pulmonary arterial lesions of tained reduction in PA pressure was also found in ambulatory patients with PAH. PDGFRa expression was found mainly PAH (MCT+PN) animals treated with PK10453. Concomi within the muscular medial layer of hypertrophied pulmonary tant with a significant reduction of PA and RV systolic pres arterioles, whereas PDGFRB and pPDGFRB were dominant Sure in these models, a reduction in RV hypertrophy and an in endothelial cells of plexiform lesions. improvement in the lumen to media ratio of pulmonary arte 0212. The selectivity of imatinib for PDGFRahas not been rioles were demonstrated. Pressure volume loops displayed previously emphasized in studies of PAH. Inhibition by ima an improvement in RV ejection fraction, a higher cardiac tinib of PDGFAA stimulated PDGFRa phosphorylation was output, and a trend towards lower stroke work in PK10453 reported to be 0.1 uM; whereas inhibition of PDGFBB stimu treated animals compared to control animals. In lung extracts lated PDGFRB phosphorylation at 0.38 uM. See, e.g., Dein of PK10453 treated animals, there was a significant reduction inger et al., “The development of imatinib as a therapeutic in the paKT(Ser473)/AKT, pSTAT3/STAT3, ppERK1/ agent for chronic myeloid leukemia.” Blood; 105:2640-53 ERK1 and pERK1/ERK1 ratios. (2005). Here, however, it was determined that, at ATPKm 0209 Because PAH is a disease substantially localized to (app), imatinib was 10 fold more selective for PDGFRO.com the lung, the hypothesis was tested that direct administration pared to the -beta receptor (ICso against PDGFRC. 71 nM. vs. of the drug to the target site via inhalation would offer the 607 nM for PDGFRB). Most PAH related cell based studies advantage of higher local concentrations (greater efficacy) interrogating the PDGFR pathway employed high doses of and lower systemic concentrations of drug (lower side imatinib (5-10 uM) and thus preclude distinction between effects). Pharmacokinetic studies demonstrated a 45 fold PDGFRC. and B receptor inhibition. advantage of inhalation delivery compared to intravenous 0213 Wu et al., “Comprehensive dissection of PDGF administration of PK10453 (Structure 2). While PK10453 PDGFR signaling pathways in PDGFR genetically defined decreased RV systolic pressure by 50% in the rat MCT model, cells. PLoS One, 3.e3794 (2008), examined PDGFR signal it did not have an adverse effect on systemic BP. Additionally, ing in genetically defined mouse embryonic fibroblasts inhaled PK10453 did not adversely affect lung function over (MEFs). The MEF's were engineered to express only the a 2-wk course. PDGFRC, PDGFRB, both or neither receptor. Signaling 0210. In the rat MCT model the present inventor compared through the PDGFRC. receptor and the PDGFRB receptor inhaled PK10453 to inhaled imatinib and found both to be were found to have both shared and distinct pathways. Thirty equally effective. These results are consistent with prior three gene sets were distinctly activated by PDGFRa and 15 reports that the PDGF receptor inhibitor imatinib, when by PDGFRB, PDGFROL/B heterodimers activated compo delivered systemically, decreased pulmonary hypertension in nents of NFKB and IL-6 signaling. Calcium flux pathways the rat MCT model. See Schermulyet al., “Reversal of experi were regulated by both PDGFRC. and PDGFRB. Signaling mental pulmonary hypertension by PDGF inhibition.”J Clin involved with angiogenesis was solely regulated via the PDG Invest, 115:2811-21 (2005). However, in the rat MCT+PN FRB pathway. This finding comports with the selective model while inhaled PK10453 was effective in lowering pull increase in phosphoPDGFRB found with neointimal lesions monary pressures, inhaled imatinib was not. The rat MCT+ of precapillary pulmonary arterioles using the MCT+PN PN model is a more aggressive model of PAH compared to the model. MCT only model, and may more accurately reflect the pathol 0214 PDGFBB has been found to induce phosphorylation ogy of the human disease. White et al., “Plexiform-like of AKT at Ser473 in pulmonary artery smooth muscle cells lesions and increased tissue factor expressionina rat model of and fibroblasts, but not pulmonary arterial endothelial cells. severe pulmonary arterial hypertension.” Am J Physiol Lung See Ogawa et al., “PDGF enhances store-operated Ca" entry Cell Mol Physiol; 293:L583-90 (2007). In vitro measurement by upregulating STIM1/Orail via activation of Akt/mTOR in of ICs for inhibition of PDGF-C. and -3 receptors showed that human pulmonary arterial Smooth muscle cells. Am J PK10453 was more potent than imatinib against both iso Physiol Cell Physiol: 302:C405-11 (2012). Increased phos forms, and that imatinib is only a modest inhibitor of the phorylation of AKT (Ser473) was also found in cells with a PDGFRB isoform Immunohistochemistry demonstrated that Smooth muscle phenotype from endarterectomies of patients US 2015/0353527 A1 Dec. 10, 2015 30 with chronic thromboembolic pulmonary arterial hyperten determined that the neointimal proliferative grade 1-2 lesions sion. See Ogawa et al., “Inhibition of mTOR attenuates store contained myofibroblasts and endothelial cells. In advanced operated Ca"entry in cells from endarterectomized tissues of grade 2 lesions there was fibrotic replacement of the vessel patients with chronic thromboembolic pulmonary hyperten media. The origin of myofibroblasts in these lesions is not sion.” Am J Physiol Lung Cell Mol Physiol; 297:L666-76 entirely clear. They could originate from infiltration of peri (2009). PDGFBB stimulation increased store operated cal vascular fibroblasts or pericytes, from circulating stem cells, cium entry via the AKT/mTOR pathway in these cells. Seeid. resident progenitor cells, or as a consequence of endothelial 0215. In pulmonary artery smooth muscle cells from con mesenchymal transition. See Yeager et al., “Progenitor cells trol and monocrotaline treated rats, however, imatinib (0.1 in pulmonary vascular remodeling.” Pulm Circ; 1:3-16 uM) decreased fetal calf serum stimulated Ser473 AKT phos (2011). While these lesions were detected, it is reasonable to phorylation, but had no effect on phosphorylation of AKT at propose that the type 1 lesion is an earlier stage lesion that can Thr30825. At this concentration it is likely that imatinib was progress to type 2 and type 3. In this model, intraluminal acting via the PDGFC. receptor. Wu et al. (2008) found that endothelial cells proliferate, transition to a myofibroblast STI-571 (imatinib) at 5 uM blocked PDGFBB stimulated AKT phosphorylation (SER473) in both PDGFRB null and phenotype (and/or the lumen is infiltrated by perivascular PDGFRC. null cell lines. The present invention included an cells/myofibroblasts) and progressively occlude the vessel ICW to examine PDGFAA and PDGFBB stimulation of AKT lumen. (Ser473) and AKT (Thr308) phosphorylation in fetal human 0218 Sakao et al., “Reversible or irreversible remodeling lung fibroblasts Inhibition by imatinib was compared to in pulmonary arterial hypertension.” Am J Respir Cell Mol PK10453 inhibition of PDGFAA or PDGFBB Stimulated Biol. 43:629-34 (2010), have highlighted the importance of AKT phosphorylation, and found that PK10453 was more distinguishing regression of vascular muscularization (re potent. verse remodeling) from potentially irreversible endothelial 0216 Nano-fluidic proteomic immunoassays, moreover, cell proliferation in PAH. The data presented here shows that were employed to quantify phosphorylated species of AKT, signaling through the PDGFRC. pathway plays an important STAT3 and ERK1/2 in lung extracts of MCT+PN animals. A role in vascular remodeling of larger pulmonary arterioles in significant reduction of phospho-AKT (Ser473), phospho PAH, whereas the PDGFRB pathway is more important in the STAT3 and ppERK1/ERK and pERK1/ERK1 in the proliferative neointimal lesions of precapillary pulmonary PK10453 treated groups was found as compared to vehicle. Schermuly et al. (2008) demonstrated a reduction in arterioles. Targeting the PDGFRB pathway with a PDGFR pERK1/2 by imatinib in the rat MCT model of PAH. Jasmin inhibitor that potently blocks this isoform (more potently than et al., “Short-term administration of a cell-permeable caveo imatinib) may influence progression of these lesions. If Such lin-1 peptide prevents the development of monocrotaline lesions are therefore treated before full fibrotic replacement induced pulmonary hypertension and right ventricular hyper and vessel regression reversibility of these lesions may exist. trophy.” Circulation, 114:912-20 (2006), have shown 0219. In conclusion, an inhaled, non-selective PDGF activation of STAT3 in the rat MCT model, and Masri et al., receptor inhibitor, PK10453 (Structure 2), was effective in “Hyperproliferative apoptosis-resistant endothelial cells in both the MCT, and MCT+PN rat models of PAH. Treatment idiopathic pulmonary arterial hypertension.” Am J Physiol with PK10453 was associated with a significant reduction in Lung Cell Mol Physiol; 293:L548-54 (2007), found that pulmonary arterial pressures in ambulatory animals, an STAT3 was activated in human idiopathic PAH. The nanof improvement in right ventricular function, and a reduction in luidic proteomic immunoassays of the present invention were RV hypertrophy. Histologic analysis demonstrated an previously used to examine the effects of imatinib on pSTAT3, and pERK 1/2 in chronic myelogenous leukemia improvement in the pulmonary arteriole lumento media ratio (CML). See Fan et al., “Nanofluidic proteomic assay for in animals treated with PK10453 and a decrease in the phos serial analysis of oncoprotein activation in clinical speci phorylation state of AKT (Ser473), STAT3 and ERK1. There mens.” Nature medicine, 15:566-71 (2009). This assay has was no significant effect of PK10453 (Structure 2) on sys utility in distinguishing monophosphorylated isoforms and temic blood pressure, and no adverse effect of PK10453 on diphosphorylated isoforms of proteins. For example, patients lung function. In contrast to imatinib, PK10453 is not selec with CML who responded to imatinib had a distinct reduction tive for the PDGFRa receptor, but rather is highly potent in levels of monophosphorylated ERK214. Here, the ERK1 against both the PDGFRC. and B isoforms. Because the PDG isoform and both the diphosphorylated form of ERK1 and the FRB pathway is more highly activated than the PDGFRC. monophosphorylated form of ERK1 predominated in lungs receptor in plexiformlesions of PAH, a non-selective PDGFR of MCT pneumonectomized rats. Treatment with PK10453 inhibitor, e.g., PK10453, thus possesses efficacy against PAH significantly decreased ppERK1/ERK and pPRK1/ERK1. and related diseases and disease pathways. 0217 Occlusion analyses were performed in accordance with the method of Homma et al., “Involvement of RhoA/Rho What is claimed is: kinase signaling in protection against monocrotaline-induced pulmonary hypertension in pneumonectomized rats by dehy 1. A method of non-selective kinase receptor inhibition for droepiandrosterone.” Am J Physiol Lung Cell Mol Physiol; treating pulmonary disorders in a Subject, comprising: 295:L71-8 (2008). In the rat MCT+PN model, the higher dose administering to the subject a therapeutically effective of inhaled PK10453 was associated with fewer Grade 2 amount of a compound of Structure 1, a tautomer, enanti occlusive lesions. These lesions were then characterized by omer, isomeror Stereoisomer of the compound, a pharmaceu immunohistochemistry with markers for vascular Smooth tically acceptable salt of the compound, tautomer, enanti muscle cells, and endothelial cells, and performed trichrome omer, isomer or Stereoisomer of the compound, or any stains to differentiate muscular from fibrotic lesions. It was mixtures thereof, wherein Structure 1 has the formula: US 2015/0353527 A1 Dec. 10, 2015 31
-continued Structure 1 HN R1 R2 N 21 O, H -Q, N r N X 2- N R3 s s’ 2 R - N2i nsR Q' and Q are independently selected from the group con R6 sisting of a direct bond, —CH, -OH, -O-CH C. N. C(=O)–C–C, C. N. C(=O)–C F, and the following group B structures: and wherein X is independently selected from carbon or nitrogen; R" and R may be the same or different and are indepen o1 o1 dently selected from the group consisting of H, C, N, O, s 1. S., Cl, Br, F. I. —CN, NO. —OH, -CH, —CF, O —C N C - groups, —C N—C(=O) groups, Substituted and unsubstituted amidinyl groups, Substi tuted and unsubstituted guanidinyl groups, Substituted 1. and unsubstituted aryl groups, Substituted and unsubsti O OH tuted alkenyl groups, Substituted and unsubstituted alky- 2 2 nyl groups; Substituted and unsubstituted heterocyclyl Q-, Q-, groups, Substituted and unsubstituted aminoalkyl groups: R. R. R. and R', may be the same or different and are independently selected from the group consisting of o1 absent H, Cl, Br, F, I, —CN, NO. —OH, -CH OH N 2. and —CF —NH, —C=N. —C—N groups, —C N— 21 C - groups, —C N C(=O)—groups, —C N—C 1 (=O) C F. —C N—C(=O)—C—C, substituted N-N and unsubstituted alkyl groups, Substituted and unsub- C-N-C(=O)-C=C. stituted aryl groups, Substituted and unsubstituted het erocyclyl groups, alkoxy groups, aryloxy groups, Sub- OH stituted and unsubstituted heterocyclylalkyl groups, Substituted and unsubstituted aminoalkyl groups, —C(=O)H, —(=O)-alkyl groups, —C(=O)-aryl groups. (=O)C)-alkyl groups, —(=O)C-aryl groups, —(=O)NH, substituted and unsubstituted alkoxyalkyl groups, Substituted and unsubstituted aryloxyalkyl and groups, and Substituted and unsubstituted heterocycly- Q is selected from the group consisting of absent, a direct loxyalkyl groups, —NH(alkyl) groups, —NH(aryl) bond, H, C, Cl, Br. F. I. —CN, NO. —CH, —CF, groups, —N(alkyl) groups, —N(aryl) groups, NH, C(=O)—, C. N. R', C=N, 3 —N(alkyl)(aryl) groups, —NH(heterocyclyl) groups, C N-C groups, —C N-C(=O)— groups, —N(heterocyclyl)(alkyl) groups, N(heterocyclyl) C. N. C(=O) C F. C. N. C(=O) (aryl) groups, and —N(heterocyclyl) groups: C=C. —C=N groups, —C(=O)— groups, R is selected from the group consisting of Q. —C(=O)—C-groups, —C(=O)C=C. —CF, C=N. —C N C groups, —C N C(=O) groups, C. N. C(=O) C F. C. N. C HN (=O)—C=C. —OH, alkoxy groups, alkoxy groups, aryloxy groups, methoxy groups, dimethoxy groups, methoxy phenol, methoxy phenol groups, dimethoxy phenol, dimethoxyphenol groups, dimethoxybenzene, Sn dimethoxy benzene groups, Substituted and unsubsti N s tuted alkyl groups, Substituted and unsubstituted aryl HN groups, and Substituted and unsubstituted heterocyclyl groups, and combinations thereof. H O and 2.-4. (canceled) 5. The method of claim 1, wherein the compound of Struc 4ns-N ture 1 is a compound of Structure 2, Structure 2a, Structure 3, / \, Structure 4 or Structure 5 as shown in the following Groun C Structures: US 2015/0353527 A1 Dec. 10, 2015 32
6. The method of claim 1, wherein the compound of Struc ture 1 is administered orally, intravenously, Subcutaneously, Structure 2 transdermally, intraperitoneally, or by inhalation. o1 7. The method of claim 1, wherein the kinase receptor is a receptor tyrosine kinase (RTK), and wherein the RTK is OH platelet derived growth factor receptor (PDGFR). H 8. The method of claim 7, wherein the PDGFR is platelet N N derived growth factor receptor-alpha (PDGFR-C) or platelet HN n derived growth factor receptor-beta (PDGFR-B) or both. 2 21 O N 9. The method of claim 7, wherein the PDGFR is a homodimer or heterodimer selected from the group consist Sa ing of PDGFR-O.C., PDGFR-?3(Band PDGFR-CfB, or any com N bination thereof. Structure 2a 10. The method of claim 7, wherein the inhibition of the PDGFR is effective in treating the pulmonary disorder, o1 wherein the pulmonary disorder is pulmonary arterial hyper o1 tension (PAH), PAH associated with plexiform and/or neoin timal lesions, PAH associated with pulmonary fibrosis and/or H N N progressive vasodegeneration, abnormal fibroblast and/or HN n myofibroblast proliferation, or pulmonary vascular disorders 2 associated with abnormal endothelial cell proliferation, or 21 O N any combination thereof. S. 11. The method of claim 7, wherein the inhibition is a N combined inhibition of both the PDGFR-O. and the PDGFR Structure 3 B. 12. The method of claim 11 wherein the inhibition prevents o1 activation of both the PDGFR-C. and the PDGFR-B by modu OH lating cognate substrate interactions. 13. The method of claim 12, wherein the cognate substrate H N N is selected from the group consisting of PDGFAA, PDGFBB HN s and PDGFAB, or any combination thereof. 22 14. The method of claim 1, wherein the pulmonary disorder N2 O N is selected from the group consisting of pulmonary arterial hypertension (PAH), PAH associated with plexiform and/or -'sN neointimal lesions, PAH associated with pulmonary fibrosis N2 and/or progressive vasodegeneration, abnormal fibroblast Structure 4 and/or myofibroblast proliferation, pulmonary vascular dis orders associated with abnormal endothelial cell, prolifera O1 tion, primary PAH, idiopathic PAH, heritable PAH, refractory OH PAH, BMPR2, ALK1, endoglin associated with hereditary H hemorrhagic telangiectasia, endoglin not associated with N N HN n hereditary hemorrhagic telangiectasia, drug-induced PAH, a toxin-induced PAH, PAH associated with systemic sclerosis, O N mixed connective tissue disease, and portal hypertension and/ H or wherein the PAH is secondary to pulmonary hypertension, a S N A w congenital heart disease, hypoxia, chronic hemolytic anemia, O newborn persistent pulmonary hypertension, pulmonary Structure 5 veno-occlusive disease (PVOD), pulmonary capillary hemangiomatosis (PCH), left heart disease pulmonary hyper o1 tension, systolic dysfunction, diastolic dysfunction, Valvular disease, lung disease, interstitial lung disease, pulmonary fibrosis, Schistosomiasis, chronic obstructive pulmonary dis H ease (COPD), sleep-disordered breathing, alveolar hypoven N N tilation disorders, chronic exposure to high altitude, develop HN n mental abnormalities, chronic thromboembolic pulmonary 2 hypertension (CTEPH), pulmonary hypertension with 21 O N unclear multifactorial mechanisms, hematologic disorders, myeloproliferative disorders, splenectomy, systemic disor Sa N ders, sarcoidosis, pulmonary Langerhans cell histiocytosis, lymphangioleimoyomatosis, neurofibromatosis, vasculitis, Group C Structures metabolic disorders, glycogen storage disease, Gaucher dis ease, thyroid disorders, tumoral obstruction, fibrosing medi astinitis, and chronic renal failure on dialysis. US 2015/0353527 A1 Dec. 10, 2015
15.-16. (canceled) -continued 17. The method of claim 1, wherein the pulmonary disorder Structure 3 is associated with abnormal: right ventricular systolic pres o1 sure (RVSP); pulmonary pressure; OH cardiac output; right ventricular (RV) hypertrophy; and/or H pulmonary arterial (PA) hypertrophy. N N HN n 18. The method of one of claims 1-17, wherein the com 2 pound of Structure 1 possesses an ICs of less than 300 nM for N21 O N the kinase receptor. 2 19.-20. (canceled) N ->N2 21. The method of any one of claim 1, wherein the inhibi Structure 4 tion occurs through a covalent or non-covalent interaction. O1 22. A method of treating pulmonary arterial hypertension OH (PAH) in a subject in need thereof, comprising: modulating H the phosphorylation-state of one or more downstream targets HN n of platelet derived growth factor receptor-alpha (PDGFR-O.) 1O. N a or platelet derived growth factor receptor-beta (PDGFR-B) or O N H both, wherein the downstream target is any substrate phos an S1 N phorylated as a result of the PDGFR-C. and/or the PDGFR-?3 A w activation, and further wherein the modulation occurs by O O administering to the Subject a compound of Structure 1, a Structure 5 tautomer, enantiomer, isomer or stereoisomer of the com o1 pound, a pharmaceutically acceptable salt of the compound, tautomer, enantiomer, isomer or stereoisomer of the com pound, or any mixtures thereof, wherein the downstream H target is selected from the group consisting of AKT, PDGFR, N N STAT3, ERK1 and ERK2, or any other downstream target of HN C n the PDGFR-O. and/or the PDGFR-B, and wherein the com 2 pound of Structure 1 is a compound of Structure 2, Structure 21 O N 2a, Structure 3, Structure 4 or Structure 5 as shown in the Sa following Group C structures: N Group C Structures
Structure 2 23. (canceled) 24. The method of claim 22, wherein the modulation is a o1 decrease of phosphorylated STAT3 to total STAT3, diphos phorylated ERK1 to total ERK1, diphosphorylated ERK2 to OH total ERK2, monophosphorylated ERK1 to total ERK1, phosphorylated PDGFR to total PDGFR, or phosphorylated H AKT to total AKT, or any combination thereof, in the subject N N compared to the Subject before the administering. 25.-28. (canceled) 2 29. A compound of Structure 1, a tautomer, enantiomer, 21 O N isomer or stereoisomer of the compound, a pharmaceutically Sa accep2table salt of the compound, tautomer, enantiomer, iso N mer or Stereoisomer of the compound, or any mixtures thereof, wherein Structure 1 has the following formula: Structure 2a Structure 1 o1 RI R2 o1 21 R4 H N N - 1S s s 2 X 21 O N Sa N US 2015/0353527 A1 Dec. 10, 2015 34
and wherein X is independently selected from carbon or -continued nitrogen; HN R" and R may be the same or different and are indepen- 21 dently selected from the group consisting of H, C, N, O, N O, S., Cl, Br. F. I. —CN, NO. —OH, -CH —CF, N —C N C - groups, —C N—C(=O)— groups, 2 N Substituted and unsubstituted amidinyl groups, Substi tuted and unsubstituted guanidinyl groups, Substituted and unsubstituted aryl groups, Substituted and unsubsti wherein the structure of Q and Q are independently tuted alkenyl groups, Substituted and unsubstituted alky Selected from the group consisting of a direct bond, nyl groups, Substituted and unsubstituted heterocyclyl CH, OH, O CH, C N C(=O)—C—C, groups, Substituted and unsubstituted aminoalkyl —C N C(=O)—C—F. and the following group B groups: Structures: R. R. R. and R', may be the same or different and are independently selected from the group consisting of absent, H., Cl, Br, F, I, —CN, NO. —OH, -CH o1 o1 —CF —NH, —C=N. —C—N groups, —C N— s 1. C - groups, —C N C(=O)—groups, —C N—C O (=O) C F. —C N—C(=O)—C—C, substituted and unsubstituted alkyl groups, Substituted and unsub stituted aryl groups, Substituted and unsubstituted het 1. erocyclyl groups, alkoxy groups, aryloxy groups, Sub O OH stituted and unsubstituted heterocyclylalkyl groups, Q?, Q?, Substituted and unsubstituted aminoalkyl groups, —C(=O)H, —C(=O)-alkyl groups, —C(=O)-aryl groups, —C(=O)C)-alkyl groups, —C(=O)C-aryl o1 s 2N. and —C(=O)-aryl groups, —C(=O)NH2. —C(=O)NH OH N 2 (alkyl) groups, —C(=O)NH(aryl) groups, groups, Sub- 21 stituted and unsubstituted heterocyclylaminoalkyl N-N groups, Substituted and unsubstituted hydroxyalkyl groups, Substituted and unsubstituted alkoxyalkyl groups, Substituted and unsubstituted aryloxyalkyl OH groups, and Substituted and unsubstituted heterocycly loxyalkyl groups, —NH(alkyl) groups, —NH(aryl) groups, —N(alkyl) groups, —N(aryl) groups, —N(alkyl)(aryl) groups, —NH(heterocyclyl) groups, —N(heterocyclyl)(alkyl) groups, N(heterocyclyl) (aryl) groups, and —N(heterocyclyl) groups; wherein Q is selected from the group consisting of absent, a direct bond, H, C, Cl, Br, F, I, —CN, NO. —CH, wherein the structure of R has the following formula is —CF, NH, C(=O)13, C N R', -C=N, selected from the group consisting of: Q. —C N C groups, —C N—C(=O)-groups, C. N. C(=O) C F. C. N. C(=O) C=C. —C=N groups, —C(=O)— groups,
C. N. C(=O) C F. C. N. C(=O) C=C. —OH, alkoxy groups, alkoxy groups, aryloxy 21 groups, methoxy groups, dimethoxy groups, methoxy phenol, methoxy phenol groups, dimethoxy phenol, N N dimethoxy phenol groups, dimethoxy benzene, dimethoxy benzene groups, Substituted and unsubsti tuted alkyl groups, Substituted and unsubstituted aryl groups, and Substituted and unsubstituted heterocyclyl groups, and combinations thereof. 30. (canceled) N 31. The compound of claim 29, wherein the compound of 1.WV Structure 1 is a compound of Structure 2a, Structure 3, Struc O O ture 4 or Structure 5 as shown below in the Group C struc tures: US 2015/0353527 A1 Dec. 10, 2015
-continued Structure 2a o1 Structure 5 o1 H N N HN n H N N 2 21 O N O C n 2 Sa 21 O N N Structure 3 S. N o1 Group C Structures OH H N N HN n 32. The compound of claim 29, wherein the salt is a chlo 2 N21 O N ride, hydrochloride, Sulfate, phosphate, mesylate, bismesy late, tosylate, lactate, tartrate, malate, bis-acetate, citrate, or 2 bishydrochloride salt. N -'sN2 Structure 4 33.-37. (canceled) 38. The method of claim 1, wherein the treatment results in O1 one or more of improved exercise capacity, increased 6 OH minute walk distance, improved functional class, an improve H ment from class IV to class III, II or I, oran improvement from N N HN n class III to class II or I, or an improvement form class II to a class I, less shortness of breath, decreased hospitalization, O N decreased need for lung transplantation, decreased need for H a S N atrial septostomy, and increased longevity or overall Survival. A w O 39.-40. (canceled)