HK1091819B

HK1091819B - Substitued pyridines and pyridazines with angiogenesis inhibiting activity

Info

Publication number: HK1091819B
Application number: HK06112290.6A
Authority: HK
Inventors: P. Dumas Jacques; Kite Joe Teddy; C.E. Kluender Harold; Lee Wendy; Nagarathnam Dhanapalan; N. Sibley Robert; Su Ning; James Boyer Stephen; A. Dixon Julie
Original assignee: Bayer Healthcare Llc
Priority date: 1999-09-28
Filing date: 2006-11-08
Publication date: 2009-07-24

Description

Substituted pyridines and pyridazines having angiogenesis inhibiting activity

The present application is a divisional application of the chinese patent application entitled "substituted pyridines and pyridazines having angiogenesis inhibiting activity" filed on 26/9/2000, application No. 00816369.3.

Technical Field

The present application relates to small molecule heterocyclic drugs, and more particularly, to substituted pyridines and pyridazines having angiogenesis inhibiting activity.

Background

Angiogenesis involves the revascularization by endothelial cell precursors or angioblasts. The first vascular structure in the embryo is formed by angiogenesis. Angiogenesis involves the development of capillaries from existing blood vessels and is the primary mechanism by which blood vessels are formed in organs such as the brain and kidneys. While angiogenesis is limited to embryonic development, angiogenesis can occur in adults, for example, during pregnancy, the female cycle, or wound healing.

The major regulator of angiogenesis and vasculogenesis in embryonic development and certain angiogenesis-dependent diseases is vascular endothelial growth factor (VEGF; also known as vascular permeability factor, VPF). VEGF represents a family of mitogen isoforms that arise from alternative RNA splicing and exist as homodimers. The VEGF KDR receptor is highly specific for vascular endothelial cells (for a review see: Farrara et al Endocr. Rev.1992, 13, 18; Neufield et al FASEB J.1999, 13, 9).

VEGF expression is induced by hypoxia (Shweiki et al Nature 1992, 359, 843) and a number of different cytokines and growth factors such as interleukin-1, interleukin-6, epidermal growth factor and transforming growth factors-alpha and-beta.

To date, VEGF and VEGF family members have been reported to bind to one or more of the three transmembrane receptor tyrosine kinases (Mustonen et al j.cell.biol., 1995, 129, 895), VEGF receptor-1 (also known as flt-1 (fms-like tyrosine kinase-1)); VEGFR-2 (also known as a kinase insert domain containing receptor (KDR), a murine analog of KDR known as fetal liver kinase-1 (flk-1)); and VEGFR-3 (also known as flt-4). KDR and flt-1 have been shown to have different signalling characteristics (Waltenberger et al J.biol.chem.1994, 269, 26988; Park et al Oncogene 1995, 10, 135). Thus, KDR undergoes strong ligand-dependent tyrosine phosphorylation in intact cells, whereas flt-1 shows a weak response. Therefore, binding to KDR is necessary for the induction of a full range of VEGF-mediated biological responses.

In vivo, VEGF plays a key role in angiogenesis and induces angiogenesis and vascular permeability. Uncontrolled VEGF expression will lead to a variety of diseases characterized by abnormal angiogenesis and/or highly osmotic processes. Thus, control of the VEGF-mediated signaling cascade would provide a useful method of controlling aberrant angiogenesis and/or hyper-osmotic processes.

Angiogenesis is considered an absolute requirement for tumor growth beyond about 1-2 mm. In tumors below this limit, oxygen and nutrients can be supplied to the cells by diffusion. However, when a certain size is reached, each tumor is dependent on angiogenesis for continued growth. Tumorigenic cells in hypoxic regions respond by stimulating VEGF production, which triggers activation of resting endothelial cells to stimulate new blood vessels formation (Shweiki et al proc. nat' 1.acad. sci., 1995, 92, 768). Furthermore, VEGF production in tumor regions without any angiogenesis can proceed through the ras signaling pathway (Grugel et al J.biol.chem., 1995, 270, 25915; Rak et al Cancer Res.1995, 55, 4575). In situ hybridization experiments have demonstrated that VEGF mRNA is severely upregulated in a variety of Human tumors including lung Cancer (Mattern et al Br. J. Cancer 1996, 73, 931), thyroid Cancer (Viglietto et al Oncogene 1995, 11, 1569), breast Cancer (Brown et al Human Pathol.1995, 26, 86), gastrointestinal Cancer (Brown et al Cancer Res.1993, 53, 4727; Suzuki et al Cancer Res.1996, 56, 3004), renal and bladder Cancer (Brown et al am. J. Pathol.1993, 1431, 1255), ovarian Cancer (Olson et al Cancer Res.1994, 54, 1255), and cervical Cancer (Guidi et al J. Nat' l Cancer Inst.1995, 87, 12137), as well as angiosarcoma (Hashimo et al enc. Clostr. 1995, 78. J. enc. 1992. 73, et al enc. intracranial tumour, multilayered tumor, Philip. J. 1993, Nat. Ogli. Olymp. J. Onkol. 73, 78, 1993). Monoclonal antibodies that neutralize KDR have been shown to be effective in blocking tumor angiogenesis (Kim et al. Nature 1993, 362, 841; Rockwell et al. mol.cell.differ.1995, 3, 315).

Over-expression of VEGF, for example under extreme hypoxic conditions, can lead to angiogenesis in the eye, leading to hyperproliferation of blood vessels, and ultimately blindness. A series of such events have been observed in a variety of retinopathies, including diabetic retinopathy, ischemic retino-venous obstruction, early-maturing retinopathy (Aiello et al, New engl.j.med.1994, 331, 1480; Peer et al, lab.invest.1995, 72, 638), and age-related macular degeneration (AMD; see Lopez et al, invest.opthalmol.vis.sci.1996, 37, 855).

In Rheumatoid Arthritis (RA), pannus growth may be mediated by the production of angiogenic factors. In synovial fluid of patients with RA, the level of immunoreactive VEGF is high, whereas in synovial fluid of patients with other forms of arthritis accompanied by degenerative joint disease, the level of VEGF is low (Koch et al, j. immunol.1994, 152, 4149). Angiogenesis inhibitor AGM-170 has been shown to prevent neovascularization of the joints in the rat collagen arthritis model (Peacock et al J. Exper. Med.1992, 175, 1135).

Increased VEGF expression has also been found in skin with psoriasis, as well as in bullous diseases associated with epidermal underwater blister formation, such as bullous pemphigoid, erythema multiforme, dermatitis herpetiformis (Brown et al j. invest. dermatol.1995, 104, 744).

Because inhibition of KDR signaling will result in inhibition of VEGF-mediated angiogenesis and permeability, KDR inhibitors are useful in the treatment of diseases characterized by aberrant angiogenesis and/or hyperosmotic processes, including the aforementioned sputum diseases.

Examples of phthalazines and other fused pyridazines that are structurally similar to the compounds of the present application are disclosed in the following patents or patent applications: WO 9835958(Novartis), US 5849741, US 3753988, US 3478028 and JP 03106875. Other references relating to phthalazine are El-Feky, S.A., Baulomy, B.E., and Abd El-Sami, Z.K., Egypt.J.Chem. (1991), Volume Data 1990, 33(2), 189-197; duhault, j., Gonnard, p., and Fenard, s., bull.soc.chim.biol., (1967), 49(2), 177-; and holova, h.m. and Jr, Partyka, r.a., j.med.chem. (1969), 12, 555-. The compounds of the present invention are different from the compounds described in each of the above-mentioned documents, and only the Novartis publication describes such compounds as angiogenesis inhibitors.

As noted above, compounds that inhibit angiogenesis have utility in the treatment of a variety of different conditions, and are therefore desirable. Such substances are the subject of the present invention.

Disclosure of Invention

In its broadest aspect, the present invention relates to three groups of compounds, or pharmaceutically acceptable salts or prodrugs thereof, wherein each group of compounds overlaps in scope with the other groups of compounds. In each group of compounds, the general formula of the compounds is the same, but it should be noted that the definition of the several groups that make up the general structure in each group is somewhat different. Thus, the identified groups of compounds differ from each other, but overlap in their ranges.

The first group of compounds has the general formula shown below

Wherein

R¹And R²Together form a structure containing two T²And a T³The bridging group, together with the rings to which it is attached, forms a bicyclic ring shown by the structure

Wherein each T²Independently represent N, CH, or CG¹；

T³Representative S, O, CR⁴G¹、C(R⁴)₂Or NR³。

In the above structure, G¹Is a substituent independently selected from the group consisting of: -N (R)⁶)₂；-NR³COR⁶(ii) a Halogen; an alkyl group; a cycloalkyl group; a lower alkenyl group; lower cycloalkenyl; halogen-substituted alkyl; amino-substituted alkyl; n-lower alkylamino substituted alkyl; alkyl substituted with N, N-di-lower alkylamino; n-lower alkanoylamino substituted alkyl; hydroxy-substituted alkyl; cyano-substituted alkyl; a carboxy-substituted alkyl group; lower alkoxycarbonyl-substituted alkyl; phenyl lower alkoxycarbonyl substituted alkyl; halogen-substituted alkylamino; amino-substituted alkylamino; n-lower alkylamino-substituted alkylamino; alkylamino substituted by N, N-di-lower alkylamino; n-lower alkanoylamino substituted alkylamino; hydroxy-substituted alkylamino; cyano-substituted alkylamino; a carboxy-substituted alkylamino group; lower alkoxycarbonyl-substituted alkylamino; phenyl lower alkoxycarbonyl substituted alkanesAn amino group; -OR⁶；-SR⁶；-S(O)R⁶；-S(O)₂R⁶(ii) a Halo-lower alkoxy; a halogenated lower alkylthio group; a halogenated lower alkylsulfonyl group; -OCOR⁶；-COR⁶；-CO₂R⁶；-CON(R⁶)₂；-CH₂OR³；-NO₂(ii) a -CN; an amidino group; guanidino; a sulfo group; -B (OH)₂(ii) a Optionally substituted aryl; optionally substituted heteroaryl; an optionally substituted saturated heterocyclic group; optionally substituted saturated heterocyclylalkyl; an optionally substituted partially unsaturated heterocyclic group; optionally substituted partially unsaturated heterocyclylalkyl; -OCO₂R³(ii) a Optionally substituted heteroarylalkyl; optionally substituted heteroaryloxy; -S (O)_p(optionally substituted heteroaryl); optionally substituted heteroarylalkoxy; -S (O)_p(optionally substituted heteroarylalkyl); -CHO; -OCON (R)⁶)₂；-NR³CO₂R⁶(ii) a and-NR³CON(R⁶)₂。

Radical R³Is H or lower alkyl. R⁶Independently selected from H; an alkyl group; a cycloalkyl group; optionally substituted aryl; optionally substituted aryl lower alkyl; lower alkyl-N (R)³)₂(ii) a And lower alkyl-OH.

In the general formula (I), R⁴Is H, halogen, or lower alkyl. Subscript p is 0, 1, or 2; and X is selected from O, S, and NR³。

The linking group Y is selected from lower alkylene; -CH₂-O-；-CH₂-S-；-CH₂-NH-；-O-；-S-；-NH-；-O-CH₂-；-S(O)-；-S(O)₂-；-SCH₂-；-S(O)CH₂-；-S(O)₂CH₂-；-CH₂S(O)-；-CH₂S(O)₂-；-(CR⁴ ₂)_n-S(O)_p- (5-membered heteroaryl) - (CR)⁴ ₂)_s-; and- (CR)⁴ ₂)_n-C(G²)(R⁴)-(CR⁴ ₂)_s-. Two last are connectedIn the linking group Y, n and s are each independently 0 or an integer of 1 to 2. Substituent G²Selected from-CN, -CO₂R³、-CON(R⁶)₂and-CH₂N(R⁶)₂。

Z represents CR⁴Or N.

For rings containing A, B, D, E, and L, possible substituents G on the ring³Is indicated by the subscript q, q being 0, 1 or 2.

Substituent G³Is a monovalent or divalent group selected from: a lower alkyl group; -NR³COR⁶(ii) a A carboxy-substituted alkyl group; lower alkoxycarbonyl-substituted alkyl; -OR⁶；-SR⁶；-S(O)R⁶；-S(O)₂R⁶；-OCOR⁶；-COR⁶；-CO₂R⁶；-CH₂OR³；-CON(R⁶)₂；-S(O)₂N(R⁶)₂；-NO₂(ii) a -CN; optionally substituted aryl; optionally substituted heteroaryl; an optionally substituted saturated heterocyclic group; an optionally substituted partially unsaturated heterocyclic group; optionally substituted heteroarylalkyl; optionally substituted heteroaryloxy; -S (O)_p(optionally substituted heteroaryl); optionally substituted heteroarylalkoxy; -S (O)_p(optionally substituted heteroarylalkyl); -OCON (R)⁶)₂；-NR³CO₂R⁶；-NR³CON(R⁶)₂(ii) a And structure T²＝T²-T³A divalent bridging group. In the divalent bridging group, each T²Independently represent N, CH, or CG³'; and T³Representative S, O, CR⁴G³′、C(R⁴)₂Or NR³。G³' represents a monovalent G group as defined in any of the above³(ii) a And the terminal T of the bridging group²Is connected to L, T³And D, thereby forming a 5-membered fused ring.

In the ring shown on the left side in formula (I), A and D independently represent N or CH; b and E independently represent N or CH; and L representsN or CH; with the following conditions: a) in the ring containing A, B, D, E, and L, the total number of N atoms is 0, 1, 2, or 3; b) when L represents CH, and q ═ 0 or any G³When it is a monovalent substituent, at least one of A and D is an N atom; and c) when L represents CH, and G³Is a structure T²＝T²-T³When the bridging group is divalent, A, B, D and E are also CH.

J is selected from aryl; a pyridyl group; and a ring of a cycloalkyl group. The subscript q' represents a substituent G on ring J⁴And is 0, 1, 2, 3, 4, or 5.

Possible substituents G on the ring J⁴Is a monovalent or divalent group selected from: -N (R)⁶)₂；-NR³COR⁶(ii) a Halogen; an alkyl group; a cycloalkyl group; a lower alkenyl group; lower cycloalkenyl; halogen-substituted alkyl; amino-substituted alkyl; n-lower alkylamino substituted alkyl; alkyl substituted with N, N-di-lower alkylamino; n-lower alkanoylamino substituted alkyl; hydroxy-substituted alkyl; cyano-substituted alkyl; a carboxy-substituted alkyl group; lower alkoxycarbonyl-substituted alkyl; phenyl lower alkoxycarbonyl substituted alkyl; halogen-substituted alkylamino; amino-substituted alkylamino; n-lower alkylamino-substituted alkylamino; alkylamino substituted by N, N-di-lower alkylamino; n-lower alkanoylamino substituted alkylamino; hydroxy-substituted alkylamino; cyano-substituted alkylamino; a carboxy-substituted alkylamino group; lower alkoxycarbonyl-substituted alkylamino; phenyl lower alkoxycarbonyl substituted alkylamino; -OR⁶；-SR⁶；-S(O)R⁶；-S(O)₂R⁶(ii) a Halo-lower alkoxy; a halogenated lower alkylthio group; a halogenated lower alkylsulfonyl group; -OCOR⁶；-COR⁶；-CO₂R⁶；-CON(R⁶)₂；-CH₂OR³；-NO₂(ii) a -CN; an amidino group; guanidino; a sulfo group; -B (OH)₂(ii) a Optionally substituted aryl; optionally substituted heteroaryl; an optionally substituted saturated heterocyclic group; an optionally substituted partially unsaturated heterocyclic group; -OCO₂R³(ii) a Optionally substituted heteroarylalkyl; optionally substituted heteroaryloxy; -S (O)_p(optionally substituted heteroaryl); optionally substituted heteroarylalkoxy; -S (O)_p(optionally substituted heteroarylalkyl); -CHO; -OCON (R)⁶)₂；-NR³CO₂R⁶；-NR³CON(R⁶)₂(ii) a And a bivalent bridging group forming a fused ring connected to and adjoining the ring J, the bridging group having the following structure:

a)

wherein each T²Independently represent N, CH, or CG⁴′；T³Representative S, O, CR⁴G⁴′、C(R⁴)₂Or NR³；G⁴' represents a monovalent G group as defined in any of the above⁴(ii) a And is via the terminal atom T²And T³To ring J;

b)

wherein each T²Independently represent N, CH, or CG⁴′；G⁴' represents a monovalent G group as defined in any of the above⁴(ii) a Provided that there are at most two bridge atoms T²May be N; and is via the terminal atom T²To ring J; and

c)

wherein each T⁴、T⁵And T⁶Independently represent O, S, CR⁴G⁴′、C(R⁴)₂Or NR³；G⁴' represents a monovalent G group as defined in any of the above⁴(ii) a And is via the terminal atom T⁴Or T⁵To ring J; with the following conditions:

i) when a T⁴Is O, S, or NR³When another T⁴Is CR⁴G⁴' or C (R)⁴)₂；

ii) comprises T⁵And T⁶The bridging group of atoms may contain up to two heteroatoms O, S, or N; and

iii) in a solvent containing T⁵And T⁶In the bridging group of the atom, when a T⁵And a T⁶Is an O atom, or two T⁶Is an O atom, said O atoms are separated by at least one carbon atom.

When G is⁴Is located on ring J with a linking group- (CR)⁴ ₂)_p-adjacent alkyl, and X is NR³Wherein R is³When it is an alkyl substituent, then G⁴And alkyl substituents R on X³Can be linked to form a structure- (CH)₂)_p′A bridging group, wherein p 'is 2, 3, or 4, with the proviso that the sum of p and p' is 2, 3, or 4, thereby forming a 5, 6, or 7 membered nitrogen containing ring.

Further conditions are: 1) at G¹、G²、G³And G⁴In (2), when two radicals R are present³Or R⁶Are each alkyl and, when located on the same N atom, may be bonded via a bond, O, S, or NR³Linked to form a nitrogen-containing heterocycle having 5-7 ring atoms; and 2) when the aryl, heteroaryl, or heterocyclyl ring is optionally substituted, the ring may carry up to 5 substituents independently selected from: amino, mono-lower alkyl-substituted amino, di-lower alkyl-substituted amino, lower alkanoylamino, halogen, lower alkylLower alkyl, hydroxy, lower alkoxy, lower alkylthio, halogenated lower alkoxy, halogenated lower alkylthio, lower alkanoyloxy, -CO₂R³、-CHO、-CH₂OR³、-OCO₂R³、-CON(R⁶)₂、-OCON(R⁶)₂、-NR³CON(R⁶)₂Nitro, amidino, guanidino, mercapto, sulfo, and cyano; and 3) when any alkyl group is attached to O, S, or N, and carries a hydroxy substituent, then the hydroxy substituent is separated from the O, S, or N, to which the alkyl group is attached by at least two carbon atoms.

The second group of compounds has the general formula

Wherein

R¹And R²：

i) Independently represents H or lower alkyl;

ii) together form a bridging group of the structure

Wherein the attachment is via a terminal carbon atom;

iii) together form a bridging group of the structure

Wherein the attachment is via a terminal carbon atom;

iv) together form a bridging group of the structure

In which one or two ring units T¹Is N, and the other ring members are CH or CG¹And the attachment is via a terminal atom; or

v) together form a complex containing two Ts²And a T³The bridging group, together with the rings to which it is attached, forms a bicyclic ring shown by the structure

Wherein each T²Independently represent N, CH, or CG¹；

T³Representative S, O, CR⁴G¹、C(R⁴)₂Or NR³。

In the above bridge structure, subscript m is 0 or an integer of 1 to 4; this means that the fused rings formed may optionally carry up to 4 substituents G¹。

G¹Is a substituent independently selected from the group consisting of: -N (R)⁶)₂；-NR³COR⁶(ii) a Halogen; an alkyl group; a cycloalkyl group; a lower alkenyl group; lower cycloalkenyl; halogen-substituted alkyl; amino-substituted alkyl; n-lower alkylamino substituted alkyl; alkyl substituted with N, N-di-lower alkylamino; n-lower alkanoylamino substituted alkyl; hydroxy-substituted alkyl; cyano-substituted alkyl; a carboxy-substituted alkyl group; lower alkoxycarbonyl-substituted alkyl; phenyl lower alkoxycarbonyl substituted alkyl; halogen-substituted alkylamino;amino-substituted alkylamino; n-lower alkylamino-substituted alkylamino; alkylamino substituted by N, N-di-lower alkylamino; n-lower alkanoylamino substituted alkylamino; hydroxy-substituted alkylamino; cyano-substituted alkylamino; a carboxy-substituted alkylamino group; lower alkoxycarbonyl-substituted alkylamino; phenyl lower alkoxycarbonyl substituted alkylamino; -OR⁶；-SR⁶；-S(O)R⁶；-S(O)₂R⁶(ii) a Halo-lower alkoxy; a halogenated lower alkylthio group; a halogenated lower alkylsulfonyl group; -OCOR⁶；-COR⁶；-CO₂R⁶；-CON(R⁶)₂；-CH₂OR³；-NO₂(ii) a -CN; an amidino group; guanidino; a sulfo group; -B (OH)₂(ii) a Optionally substituted aryl; optionally substituted heteroaryl; an optionally substituted saturated heterocyclic group; optionally substituted saturated heterocyclylalkyl; an optionally substituted partially unsaturated heterocyclic group; optionally substituted partially unsaturated heterocyclylalkyl; -OCO₂R³(ii) a Optionally substituted heteroarylalkyl; optionally substituted heteroaryloxy; -S (O)_p(optionally substituted heteroaryl); optionally substituted heteroarylalkoxy; -S (O)_p(optionally substituted heteroarylalkyl); -CHO; -OCON (R)⁶)₂；-NR³CO₂R⁶(ii) a and-NR³CON(R⁶)₂。

In the general formula (I), R⁴Is H, halogen, or lower alkyl; subscript p is 0, 1, or 2; and X is selected from O, S, and NR³。

The linking group Y is selected from lower alkylene; -CH₂-O-；-CH₂-S-；-CH₂-NH-；-O-；-S-；-NH-；-O-CH₂-；-S(O)-；-S(O)₂-；-SCH₂-；-S(O)CH₂-；-S(O)₂CH₂-；-CH₂S(O)-；-CH₂S(O)₂-；-(CR⁴ ₂)_n-S(O)_p- (5-membered heteroaryl) - (CR)⁴ ₂)_s-; and- (CR)⁴ ₂)_n-C(G²)(R⁴)-(CR⁴ ₂)_s-. In the latter two linking groups Y, n and s are each independently 0 or an integer from 1 to 2. G²Selected from-CN, -CO₂R³、-CON(R⁶)₂and-CH₂N(R⁶)₂。

Z represents N or CR⁴。

For rings containing A, B, D, E, and L, possible substituents G on the ring³Is indicated by the subscript q, q being 1 or 2.

In the ring shown on the left side in formula (I), A and D independently represent CH; b and E independently represent CH; and L is CH; with the following conditions: the benzene ring formed carries the structure T²＝T²-T³Divalent bridge radical as G³And (4) a substituent.

G⁴Is a monovalent or divalent group selected from: -N (R)⁶)₂；-NR³COR⁶(ii) a Halogen; an alkyl group; a cycloalkyl group; a lower alkenyl group; lower cycloalkenyl; halogen-substituted alkyl; amino-substituted alkyl; n-lower alkylamino substituted alkyl; alkyl substituted with N, N-di-lower alkylamino; n-lower alkanoylamino substituted alkyl; hydroxy-substituted alkyl; cyano-substituted alkyl; a carboxy-substituted alkyl group; lower alkoxycarbonyl-substituted alkyl; phenyl lower alkoxycarbonyl substituted alkyl; halogen-substituted alkylamino; amino-substituted alkylamino; n-lower alkylamino-substituted alkylamino; alkylamino substituted by N, N-di-lower alkylamino; n-lower alkanoylamino substituted alkylamino; hydroxy-substituted alkylamino; cyano-substituted alkylamino; a carboxy-substituted alkylamino group; lower alkoxycarbonyl-substituted alkylamino; phenyl lower alkoxycarbonyl substituted alkylamino; -OR⁶；-SR⁶；-S(O)R⁶；-S(O)₂R⁶(ii) a Halo-lower alkoxy; a halogenated lower alkylthio group; a halogenated lower alkylsulfonyl group; -OCOR⁶；-COR⁶；-CO₂R⁶；-CON(R⁶)₂；-CH₂OR³；-NO₂(ii) a -CN; an amidino group; guanidino; a sulfo group; -B (OH)₂(ii) a Optionally substituted aryl; optionally substitutedA heteroaryl group; an optionally substituted saturated heterocyclic group; an optionally substituted partially unsaturated heterocyclic group; -OCO₂R³(ii) a Optionally substituted heteroarylalkyl; optionally substituted heteroaryloxy; -S (O)_p(optionally substituted heteroaryl); optionally substituted heteroarylalkoxy; -S (O)_p(optionally substituted heteroarylalkyl); -CHO; -OCON (R)⁶)₂；-NR³CO₂R⁶；-NR³CON(R⁶)₂(ii) a And a bivalent bridging group forming a fused ring connected to and adjoining the ring J, the bridging group having the following structure:

a)

b)

c)

i) when a T⁴Is O, S, or NR³When another T⁴Is CR⁴G⁴' or C (R)⁴)₂；

Further conditions are: 1) at G¹、G²、G³And G⁴In (2), when two radicals R are present³Or R⁶Are each alkyl and, when located on the same N atom, may be bonded via a bond, O, S, or NR³Linked to form a nitrogen-containing heterocycle having 5-7 ring atoms; and 2) when the aryl, heteroaryl, or heterocyclyl ring is optionally substituted, the ring may carry up to 5 substituents independently selected from: amino, lower alkylSubstituted amino, di-lower alkyl substituted amino, lower alkanoylamino, halogen, lower alkyl, halogenated lower alkyl, hydroxy, lower alkoxy, lower alkylthio, halogenated lower alkoxy, halogenated lower alkylthio, lower alkanoyloxy, -CO₂R³、-CHO、-CH₂OR³、-OCO₂R³、-CON(R⁶)₂、-OCON(R⁶)₂、-NR³CON(R⁶)₂Nitro, amidino, guanidino, mercapto, sulfo, and cyano; and 3) when any alkyl group is attached to O, S, or N, and carries a hydroxy substituent, then the hydroxy substituent is separated from the O, S, or N, to which the alkyl group is attached by at least two carbon atoms.

The third group of compounds has the general formula

Wherein

R¹And R²：

i) Independently represents H or lower alkyl;

ii) together form a bridging group of the structure

Wherein the attachment is via a terminal carbon atom;

iii) together form a bridging group of the structure

Wherein the attachment is via a terminal carbon atom;

iv) together form a bridging group of the structure

Wherein each T²Independently represent N, CH, or CG¹；

T³Representative S, O, CR⁴G¹、C(R⁴)₂Or NR³。

G¹Is a substituent independently selected from the group consisting of: -N (R)⁶)₂；-NR³COR⁶(ii) a Halogen; an alkyl group; a cycloalkyl group; a lower alkenyl group; lower cycloalkenyl; halogen-substituted alkyl; amino-substituted alkyl; n-lower alkylamino substituted alkyl; alkyl substituted with N, N-di-lower alkylamino; n-lower alkanoylamino substituted alkyl; hydroxy-substituted alkyl; cyano-substituted alkyl; a carboxy-substituted alkyl group; lower alkoxycarbonyl groupSubstituted alkyl; phenyl lower alkoxycarbonyl substituted alkyl; halogen-substituted alkylamino; amino-substituted alkylamino; n-lower alkylamino-substituted alkylamino; alkylamino substituted by N, N-di-lower alkylamino; n-lower alkanoylamino substituted alkylamino; hydroxy-substituted alkylamino; cyano-substituted alkylamino; a carboxy-substituted alkylamino group; lower alkoxycarbonyl-substituted alkylamino; phenyl lower alkoxycarbonyl substituted alkylamino; -OR⁶；-SR⁶；-S(O)R⁶；-S(O)₂R⁶(ii) a Halo-lower alkoxy; a halogenated lower alkylthio group; a halogenated lower alkylsulfonyl group; -OCOR⁶；-COR⁶；-CO₂R⁶；-CON(R⁶)₂；-CH₂OR³；-NO₂(ii) a -CN; an amidino group; guanidino; a sulfo group; -B (OH)₂(ii) a Optionally substituted aryl; optionally substituted heteroaryl; an optionally substituted saturated heterocyclic group; optionally substituted saturated heterocyclylalkyl; an optionally substituted partially unsaturated heterocyclic group; optionally substituted partially unsaturated heterocyclylalkyl; -OCO₂R³(ii) a Optionally substituted heteroarylalkyl; optionally substituted heteroaryloxy; -S (O)_p(optionally substituted heteroaryl); optionally substituted heteroarylalkoxy; -S (O)_p(optionally substituted heteroarylalkyl); -CHO; -OCON (R)⁶)₂；-NR³CO₂R⁶(ii) a and-NR³CON(R⁶)₂。

Z represents CR⁴。

Substituent G³Is a monovalent or divalent group selected from: -NR³COR⁶(ii) a A carboxy-substituted alkyl group; lower alkoxycarbonyl-substituted alkyl; -OR⁶；-SR⁶；-S(O)R⁶；-S(O)₂R⁶；-OCOR⁶；-COR⁶；-CO₂R⁶；-CH₂OR³；-CON(R⁶)₂；-S(O)₂N(R⁶)₂；-NO₂(ii) a -CN; optionally substituted aryl; optionally substituted heteroaryl; an optionally substituted saturated heterocyclic group; an optionally substituted partially unsaturated heterocyclic group; optionally substituted heteroarylalkyl; optionally substituted heteroaryloxy; -S (O)_p(optionally substituted heteroaryl); optionally substituted heteroarylalkoxy; -S (O)_p(optionally substituted heteroarylalkyl); -OCON (R)⁶)₂；-NR³CO₂R⁶；-NR³CON(R⁶)₂(ii) a And structure T²＝T²-T³A divalent bridging group. In the divalent bridging group, each T²Independently represent N, CH, or CG³'; and T³Representative S, O, CR⁴G³′、C(R⁴)₂Or NR³。G³' represents a monovalent G group as defined in any of the above³(ii) a And the terminal T of the bridging group²Is connected to L, T³And D, thereby forming a 5-membered fused ring.

In the ring shown on the left side in formula (I), A and D independently represent N or CH; b and E independently represent N or CH; and L represents N or CH; with the following conditions: a) in the ring containing A, B, D, E, and L, the total number of N atoms is 0, 1, 2, or 3; b) when L represents CH, and any G³When it is a monovalent substituent, at least one of A and D is an N atom; and c) when L represents CH, and G³Is a structure T²＝T²-T³When the bridging group is divalent, A, B, D and E are also CH.

G⁴Is a monovalent or divalent group selected from: -N (R)⁶)₂；-NR³COR⁶(ii) a Halogen; an alkyl group; a cycloalkyl group; a lower alkenyl group; lower cycloalkenyl; halogen-substituted alkyl; amino-substituted alkyl; n-lower alkylamino substituted alkyl; alkyl substituted with N, N-di-lower alkylamino; n-lower alkanoylamino substituted alkyl; hydroxy-substituted alkyl; cyano-substituted alkyl; a carboxy-substituted alkyl group; lower alkoxycarbonyl-substituted alkyl; phenyl lower alkoxycarbonyl substituted alkyl; halogen-substituted alkylamino; amino-substituted alkylamino; n-lower alkylamino-substituted alkylamino; alkylamino substituted by N, N-di-lower alkylamino; n-lower alkanoylamino substituted alkylamino; hydroxy-substituted alkylamino; cyano-substituted alkylamino; a carboxy-substituted alkylamino group; lower alkoxycarbonyl-substituted alkylamino; phenyl lower alkoxycarbonyl substituted alkylamino; -OR⁶；-SR⁶；-S(O)R⁶；-S(O)₂R⁶(ii) a Halo-lower alkoxy; halogenated lower alkyl sulfideA group; a halogenated lower alkylsulfonyl group; -OCOR⁶；-COR⁶；-CO₂R⁶；-CON(R⁶)₂；-CH₂OR³；-NO₂(ii) a -CN; an amidino group; guanidino; a sulfo group; -B (OH)₂(ii) a Optionally substituted aryl; optionally substituted heteroaryl; an optionally substituted saturated heterocyclic group; an optionally substituted partially unsaturated heterocyclic group; -OCO₂R³(ii) a Optionally substituted heteroarylalkyl; optionally substituted heteroaryloxy; -S (O)_p(optionally substituted heteroaryl); optionally substituted heteroarylalkoxy; -S (O)_p(optionally substituted heteroarylalkyl); -CHO; -OCON (R)⁶)₂；-NR³CO₂R⁶；-NR³CON(R⁶)₂(ii) a And a bivalent bridging group forming a fused ring connected to and adjoining the ring J, the bridging group having the following structure:

a)

b)

wherein each T²Independently represent N, CH, or CG⁴′；G⁴' represents a monovalent G group as defined in any of the above⁴(ii) a Provided that there are at most two bridge atoms T²May be N;and is via the terminal atom T²To ring J; and

c)

i) when a T⁴Is O, S, or NR³When another T⁴Is CR⁴G⁴' or C (R)⁴)₂；

iii) in a solvent containing T⁵And T⁶In the bridging group of the atom, when a T⁵And a T⁶Is an O atom, or two T⁶When is an O atom, said O atoms are separated by at least one carbon atom;

Further conditions are: 1) at G¹、G²、G³And G⁴In (2), when two radicals R are present³Or R⁶Are each an alkyl group and are located at the same NWhen on the atom, they may be bonded through a bond, O, S, or NR³Linked to form a nitrogen-containing heterocycle having 5-7 ring atoms; and 2) when the aryl, heteroaryl, or heterocyclyl ring is optionally substituted, the ring may carry up to 5 substituents independently selected from: amino, mono-lower alkyl-substituted amino, di-lower alkyl-substituted amino, lower alkanoylamino, halogen, lower alkyl, halogenated lower alkyl, hydroxy, lower alkoxy, lower alkylthio, halogenated lower alkoxy, halogenated lower alkylthio, lower alkanoyloxy, -CO₂R³、-CHO、-CH₂OR³、-OCO₂R³、-CON(R⁶)₂、-OCON(R⁶)₂、-NR³CON(R⁶)₂Nitro, amidino, guanidino, mercapto, sulfo, and cyano; and 3) when any alkyl group is attached to O, S, or N, and carries a hydroxy substituent, then the hydroxy substituent is separated from the O, S, or N, to which the alkyl group is attached by at least two carbon atoms.

Pharmaceutically acceptable salts of these compounds, as well as common prodrugs of these compounds, such as O-acyl derivatives of the compounds of the invention containing a hydroxyl group, are also within the scope of the invention.

The invention also relates to pharmaceutical compositions comprising one or more compounds of the invention, or salts or prodrugs thereof, in a pharmaceutically acceptable carrier.

The invention also relates to methods of using these compounds to treat a mammal suffering from a condition characterized by abnormal angiogenesis or hyper-osmotic processes, comprising administering to the mammal an amount of a compound of the present invention, or a salt or prodrug thereof, effective to treat the condition.

Defining:

the prefix "lower" refers to groups having up to and including up to 7 atoms, especially up to and including up to 5 carbon atoms, which groups are straight or branched having one or more branches.

"alkyl" refers to a hydrocarbon group having up to 12 carbon atoms, which may be straight or branched with one or more branches. Alkyl is especially lower alkyl.

When compounds, salts, and the like take the plural form, this also includes the singular form of compounds, salts, and the like.

Any asymmetric carbon atom may be present in the (R) -, (S) -or (R, S) -configuration, preferably in the (R) -or (S) -configuration. Substituents on double bonds or rings may be present in the cis- (═ Z-) or trans- (═ E-) form. Thus, the compounds may exist as mixtures of isomers or as pure isomers, preferably as pure enantiomers, diastereomers and isomers with pure cis-or trans-double bonds.

The lower alkylene radical Y may be branched or unbranched, preferably unbranched, especially methylene (-CH)₂) Ethylene (-CH)₂-CH₂)1, 3-propylene (-CH)₂-CH₂-CH₂) Or 1, 4-butylene (-CH)₂CH₂CH₂CH₂). When Y is lower alkylene, it is most preferably methylene.

"aryl" means an aromatic group having 6 to 14 carbon atoms, such as phenyl, naphthyl, fluorenyl, or phenanthryl.

"halogen" means fluorine, chlorine, bromine, or iodine, but especially fluorine, chlorine, or bromine.

"pyridyl" means 1-pyridyl, 2-pyridyl, or 3-pyridyl, but especially 2-pyridyl or 3-pyridyl.

"cycloalkyl" is a saturated carbocyclic ring containing 3 to 12 carbon atoms, but preferably 3 to 8 carbon atoms.

"cycloalkenyl" refers to a non-reactive, non-aromatic unsaturated carbocyclic ring containing 3 to 12 carbon atoms, but preferably 3 to 8 carbon atoms, and up to 3 double bonds. It is well known to those skilled in the art that cycloalkenyl groups that differ from aryl groups by only one double bond, such as cyclohexadiene, are not suitable drugs because they are not sufficiently non-reactive to be useful as substituents outside the scope of the present invention.

Cycloalkyl and cycloalkenyl groups may contain branching points such that they may be substituted with alkyl or alkenyl groups. Examples of such branched cyclic groups are 3, 4-dimethylcyclopentyl, 4-allylcyclohexyl or 3-ethylcyclopent-3-enyl.

Salts are in particular pharmaceutically acceptable salts, for example acid addition salts, of the compounds of the formula I, preferably salts of the compounds of the formula I having a basic nitrogen atom with organic or inorganic acids. Suitable inorganic acids are, for example, hydrohalic acids such as hydrochloric acid, sulfuric acid, or phosphoric acid. Suitable organic acids are, for example, carboxylic acids, phosphonic acids, sulfonic acids, or sulfamic acids, such as acetic acid, propionic acid, octanoic acid, decanoic acid, lauric acid, glycolic acid, lactic acid, hydroxybutyric acid, gluconic acid, glucomonocarboxylic acid, fumaric acid, succinic acid, adipic acid, pimelic acid, suberic acid, azelaic acid, malic acid, tartaric acid, citric acid, glucaric acid, galactaric acid, amino acids, such as glutamic acid, aspartic acid, N-methylglycine, acetylaminoacetic acid, N-acetylasparagine or N-acetylcysteine, pyruvic acid, acetoacetic acid, phosphoserine, 2-glycerophosphoric acid, or 3-glycerophosphoric acid.

In the definition of Y, the "diyl" - (5-membered heteroaryl) - "means a 5-membered aromatic heterocyclic ring containing 1 to 3 hetero atoms selected from O, S, and N, wherein the number of N atoms is 0 to 3, the number of O and S atoms is each 0 to 1, and the aromatic heterocyclic ring is bonded to sulfur via carbon, and to- (CR) via a C or N atom⁴ ₂)_s-connecting. Examples of such diradicals include

At G¹、G²、G³And G⁴In the definition of (1), it is stated that when two radicals R are present³Or R⁶When located on a single N, they may be linked to form a heterocyclic ring having from 5 to 7 atoms. Containing N to which they are attachedExamples of heterocycles of (a) are:

"Heterocyclyl" or "heterocycle" means a 5-7 membered heterocyclic ring system having 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur, which heterocyclic ring system may be unsaturated or fully or partially saturated, and unsubstituted or substituted, especially by lower alkyl, such as methyl, ethyl, 1-propyl, 2-propyl, or tert-butyl.

When it is indicated that the aryl, heteroaryl, or heterocyclyl group may be optionally substituted, the ring may carry up to 5 substituents independently selected from: amino, mono-lower alkyl-substituted amino, di-lower alkyl-substituted amino, lower alkanoylamino, halogen, lower alkyl, halogenated lower alkyl such as trifluoromethyl, hydroxy, lower alkoxy, lower alkylthio, halogenated lower alkoxy such as trifluoromethoxy, halogenated lower alkylthio such as trifluoromethylthio, lower alkanoyloxy, -CO₂R³、-CHO、-CH₂OR³、-OCO₂R³、-CON(R⁶)₂、-OCON(R⁶)₂、-NR³CON(R⁶)₂Nitro, amidino, guanidino, mercapto, sulfo, and cyano.

In the ring attached to Y, ring units A, B, D, E, and L may be N or CH, with the understanding that the optional substituent G³Must be attached to carbon, not to nitrogen, and when a given carbon atom carries a substituent G³When G is above³Is instead of the absence of G³When the carbon carries an H atom. Examples of rings J and two adjacent G which may together form a second fused ring are:

"heteroaryl" refers to a monocyclic or fused bicyclic aromatic system having a total of 5 to 10 atoms, 1 to 4 of which are heteroatoms selected from nitrogen, oxygen, and sulfur, with the remaining atoms being carbon. Heteroaryl groups are preferably monocyclic systems having a total of 5 or 6 atoms, of which 1 to 3 are heteroatoms.

"alkenyl" means an unsaturated group having up to 12 carbon atoms, which may be straight-chain or branched with one or more branches, and containing up to 3 double bonds. Alkenyl is especially lower alkenyl having up to 2 double bonds.

"alkanoyl" refers to alkylcarbonyl, especially lower alkylcarbonyl.

Halogenated lower alkyl, halogenated lower alkoxy and halogenated lower alkylthio refer to substituents wherein alkyl is partially or fully substituted by halogen, preferably by chlorine and/or fluorine, most preferably by fluorine. Examples of such substituents are trifluoromethyl, trifluoromethoxy, trifluoromethylthio, 1, 2, 2-tetrafluoroethoxy, dichloromethyl, fluoromethyl and difluoromethyl.

When a substituent is named as a string of groups such as "phenyl lower alkoxycarbonyl substituted alkylamino", it is understood that the point of attachment is on the last part of the string (amino for the above example) and that the other groups of the string are attached to each other in the order in which they are listed in the string. Thus, examples of "phenylamino substituted by phenyl lower alkoxycarbonyl" are:

when the substituent is present as a chain of radicals having a bond at the start (generally indicated by dashes), e.g. "S (O)_p(optionally substituted heteroarylalkyl) "nomenclature, it is understood that the point of attachment is at the first atom (S or sulfur for the above examples) of the series,and the other groups of the series are linked to each other in the order in which they are listed in the series. Thus, example "-S (O)_p(optionally substituted heteroarylalkyl) "is:

it will be appreciated that the leftmost part of each variable of the linking group Y is attached to the ring containing A, B, D, E, and L, and the rightmost part of the linking group is attached to the pyridazine moiety of formula (la). Thus, a linking group "-CH is used₂-O- "or a linking group" -O-CH₂Examples of- "represent the following compounds of the invention:

in the general formula (I), preferred and most preferred groups are as follows.

R¹And R²Preferably:

i) together form a bridging group of the structure

Wherein the attachment is via a terminal carbon atom;

ii) together form a bridging group of the structure

Wherein one or two rings are singlyYuan T¹Is N and the other ring unit is CH, and the connection is via a terminal atom; or

iii) together form a complex containing two Ts²And a T³The bridging group, together with the rings to which it is attached, forms a bicyclic ring shown by the structure

Wherein each T²Independently represent N, CH, or CG¹；

T³Representative S, O, CH₂Or NR³(ii) a And is

With the following conditions: when T is³When is O or S, at least one T²Is CH or CG¹。

Most preferably, any G¹Located on the non-terminal atom of the bridging group. Most preferably, in the bridging group of iii), the terminal T²Is N or CH, not terminal T²Is CH or CG¹And T is³Is S or O.

The subscript m is preferably 0 or an integer of 1 to 2, and the substituent G¹Preferably selected from: -N (R)⁶)₂；-NR³COR⁶(ii) a Halogen; a lower alkyl group; hydroxy-substituted alkyl; amino-substituted alkylamino; n-lower alkylamino-substituted alkylamino; alkylamino substituted by N, N-di-lower alkylamino; hydroxy-substituted alkylamino; a carboxy-substituted alkylamino group; lower alkoxycarbonyl-substituted alkylamino; -OR⁶；-SR⁶；-S(O)R⁶；-S(O)₂R⁶(ii) a Halo-lower alkoxy; a halogenated lower alkylthio group; a halogenated lower alkylsulfonyl group; -OCOR⁶；-COR⁶；-CO₂R⁶；-CON(R⁶)₂；-NO₂(ii) a -CN; optionally substituted heteroarylalkyl; optionally substituted heteroaryloxy; optionally substitutedHeteroarylalkoxy; and-S (O)_p(optionally substituted heteroarylalkyl). Most preferably, m is 0, and G¹Is a substituent independently selected from the group consisting of: -N (R)⁶)₂；-NR³COR⁶(ii) a Halogen; -OR⁶Wherein R is⁶Represents lower alkyl; -NO₂(ii) a Optionally substituted heteroaryloxy; and optionally substituted heteroarylalkoxy.

When R is⁶When it is an alkyl group, it is preferably a lower alkyl group. Radical R⁴Preferably H; p is preferably 0 or 1; and X is preferably NR³。

In the linking group Y, the subscripts n and s are preferably 0 or 1, most preferably 0. Y is preferably selected from lower alkylene; -CH₂-O-；-CH₂-S-；-CH₂-NH-；-S-；-NH-；-(CR⁴ ₂)_n-S(O)_p- (5-membered heteroaryl) - (CR)⁴ ₂)_s-；-(CR⁴ ₂)_n-C(G²)(R⁴)-(CR⁴ ₂)_s-; and-O-CH₂-. Y is most preferably selected from-CH₂-O-；-CH₂-NH-；-S-；-NH-；-(CR⁴ ₂)_n-S(O)_p- (5-membered heteroaryl) - (CR)⁴ ₂)_s-; and-O-CH₂-。

In the ring to the left of structure (I), A, D, B, and E are preferably CH, and L is N or CH, provided that: when L is N, any substituent G³Preferably a monovalent group, any substituent G when L is CH³Divalent groups are preferred.

Substituent G³Preferably selected from monovalent lower alkyl groups; -NR³COR⁶；-OR⁶；-SR⁶；-S(O)R⁶；-S(O)₂R⁶；-CO₂R⁶；-CON(R⁶)₂；-S(O)₂N(R⁶)₂(ii) a -CN; optionally substituted aryl; optionally substituted heteroaryl; optionally substituted heteroarylalkyl; optionally substituted heteroaryloxy; -S (O)_p(optionally takingSubstituted heteroaryl); optionally substituted heteroarylalkoxy; -S (O)_p(optionally substituted heteroarylalkyl); and structure T²＝T²-T³A divalent bridging group of (1), wherein T²Represents N or CH. T is³Preferably S, O, CR⁴ ₂Or NR³。

G³Most preferably selected from monovalent lower alkyl groups; -NR³COR⁶；-CO₂R⁶；-CON(R⁶)₂；-S(O)₂N(R⁶)₂(ii) a And structure T²＝T²-T³A divalent bridging group of (1), wherein T²Represents N or CH. T is³Most preferably S, O, CH₂Or NR³。

Represents a substituent G³The number of subscripts q is most preferably 1.

Ring J is preferably a phenyl ring, representing the substituent G on the phenyl ring⁴The number subscript q' is preferably 0, 1, 2, or 3. The subscript q' is most preferably 1, or 2.

G⁴Preferably selected from: -N (R)⁶)₂；-NR³COR⁶(ii) a Halogen; an alkyl group; halogen-substituted alkyl; hydroxy-substituted alkyl; a carboxy-substituted alkyl group; lower alkoxycarbonyl-substituted alkyl; amino-substituted alkylamino; n-lower alkylamino-substituted alkylamino; alkylamino substituted by N, N-di-lower alkylamino; n-lower alkanoylamino substituted alkylamino; hydroxy-substituted alkylamino; a carboxy-substituted alkylamino group; lower alkoxycarbonyl-substituted alkylamino; phenyl lower alkoxycarbonyl substituted alkylamino; -OR⁶；-SR⁶；-S(O)R⁶；-S(O)₂R⁶(ii) a Halo-lower alkoxy; a halogenated lower alkylthio group; a halogenated lower alkylsulfonyl group; -OCOR⁶；-COR⁶；-CO₂R⁶；-CON(R⁶)₂；-CH₂OR³；-NO₂(ii) a -CN; optionally substituted heteroarylalkyl; optionally substituted heteroaryloxy; -S (O)_p(optionally substituted heteroaryl); optionally substituted heteroarylalkoxy; -S (O)_p(optionally substituted heteroarylalkyl); and a fused ring-forming bridging group attached to and connecting adjacent positions of the phenyl ring, said bridging group having the structure:

a)

wherein each T²Independently represents N, or CH; t is³Represents S, or O; and is via the terminal atom T²And T³Is connected with a phenyl ring;

b)

wherein each T²Independently represent N, CH, or CG⁴'; provided that there are at most two bridge atoms T²May be N; and is via the terminal atom T²Is connected with a phenyl ring; and

c)

wherein each T⁵And T⁶Independently represent O, S, or CH₂(ii) a And is via the terminal atom T⁵Is connected with a phenyl ring; with the following conditions:

i) comprising T⁵And T⁶The bridging group of atoms may contain up to two heteroatoms O, S, or N; and

ii) in a solvent system comprising T⁵And T⁶Of atomsIn the bridging group, when a T⁵And a T⁶Is an O atom, or two T⁶Is an O atom, said O atoms are separated by at least one carbon atom.

Form G⁴All or a part of the alkyl groups are preferably lower alkyl groups.

When G is⁴Is located on ring J with a linking group- (CR)⁴ ₂)_p-adjacent alkyl, and X is NR³Wherein R is³When it is an alkyl substituent, then G⁴And alkyl substituents R on X³Can be linked to form a structure- (CH)₂)_p′A bridging group, wherein p 'is preferably 2 or 3, with the proviso that the sum of p and p' is 2 or 3, thereby forming a nitrogen-containing 5-or 6-membered ring. Most preferably, p and p' sum to 2, thereby forming a 5-membered ring.

Most preferably, at G¹、G²、G³And G⁴In (2), when two radicals R are present⁶Are each alkyl and, when located on the same N atom, may be bonded via a bond, O, S, or NR³Linked to form a nitrogen-containing heterocycle having 5-6 ring atoms.

Preferably, when the aryl, heteroaryl, or heterocyclyl ring is optionally substituted, the ring may carry up to 2 substituents independently selected from: amino, mono-lower alkyl-substituted amino, di-lower alkyl-substituted amino, lower alkanoylamino, halogen, lower alkyl, halogenated lower alkyl, hydroxy, lower alkoxy, lower alkylthio, halogenated lower alkoxy, halogenated lower alkylthio, -CH₂OR³Nitro, and cyano.

The methods of the invention are useful for treating VEGF-mediated disorders in humans and other mammals.

The compounds may be administered orally, transdermally, parenterally, by injection, by inhalation or spray, or sublingually, rectally, or vaginally in dosage unit formulations. The term "administration by injection" includes intravenous, intra-articular, intramuscular, subcutaneous and parenteral injection, as well as the use of infusion techniques. Transdermal administration may include topical administration or transdermal administration. The compound or compounds may be present together with one or more non-toxic pharmaceutically acceptable carriers and, if desired, other active ingredients.

Compositions for oral administration may be prepared by any suitable method known in the art for preparing pharmaceutical compositions. Such compositions may contain one or more substances selected from diluents, sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide a palatable preparation.

Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients may be, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as corn starch, or alginic acid; and binding agents, such as magnesium stearate, stearic acid or talc. The tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby achieve a sustained action over a longer period. For example, a time delay material such as glycerol monostearate or glycerol distearate may be employed. These compounds can also be formulated in solid, immediate release forms.

Formulations for oral administration may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin; or in the form of soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions containing the active ingredient in admixture with excipients suitable for the manufacture of aqueous suspensions may also be employed. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents may be a naturally-occurring phosphatide, for example lecithin, or condensation products of ethylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethylene oxide cetyl alcohol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives, for example ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.

By the addition of water, dispersion powders and granules suitable for the preparation of aqueous suspensions may provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are those enumerated above. Other excipients, for example sweetening, flavoring and coloring agents, may also be present.

The compounds may also be in the form of non-aqueous liquid preparations, for example, oil suspensions which may be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or arachis oil, or in a mineral oil, for example liquid paraffin. The oil suspension may contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and flavoring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an antioxidant such as ascorbic acid.

The pharmaceutical compositions of the present invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, for example olive oil or arachis oil, or a mineral oil, for example liquid paraffin, or mixtures thereof. Suitable emulsifying agents may be naturally-occurring gums, for example gum acacia or gum tragacanth, naturally-occurring phosphatides, for example soya bean, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example sorbitan monooleate, and condensation products of the said partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening and flavoring agents.

Syrups and elixirs may also be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative and flavoring and coloring agents.

The compounds of the present invention may also be administered in the form of suppositories for rectal or vaginal administration. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal or vaginal temperatures and will therefore melt in the rectum or vagina to release the drug. Such materials include coconut oil and polyethylene glycols.

The compounds of the present invention can also be administered transdermally using methods known to those skilled in the art (see, e.g., Chien; "Transdermal Controlled systems; Marcel Dekker. Inc.; 1987.Lipp et al WO 94/04157, 3Mar 94). For example, a solution or suspension of a compound of formula I in a suitable volatile solvent, optionally containing a penetration enhancer, may be mixed with other additives known to those skilled in the art, such as matrix materials and biocides. After sterilization, the resulting mixture may be formulated into dosage forms according to known methods. Additionally, solutions or suspensions of the compounds of formula I can be formulated as lotions or ointments by treatment with emulsifiers and water.

Suitable solvents for preparing transdermal delivery systems are known to those skilled in the art and include lower alcohols such as ethanol or isopropanol, lower ketones such as acetone, lower carboxylic acid esters such as ethyl acetate, polar ethers such as tetrahydrofuran, lower hydrocarbons such as hexane, cyclohexane or benzene, or halogenated hydrocarbons such as dichloromethane, chloroform, trichlorotrifluoroethane or trichlorofluoroethane. Suitable solvents may also include mixtures of one or more selected from the following: lower alcohols, lower ketones, lower carboxylic acid esters, polar ethers, lower hydrocarbons, halogenated hydrocarbons.

Penetration enhancers suitable for use in transdermal delivery systems are known to those skilled in the art and include, for example, mono-or polyhydric alcohols such as ethanol, propylene glycol or benzyl alcohol, saturated or unsaturated C₈-C₁₈Fatty alcohols, e.g. lauryl or cetyl alcohol, saturated or unsaturated C₈-C₁₈Fatty acids, such as stearic acid, saturated or unsaturated fatty esters having up to 24 carbon atoms, such as the methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl or monoglycerides of the following acids: acetic acid, caproic acid, lauric acid, myristic acid, stearic acid, or palmitic acid, or diesters of saturated or unsaturated dicarboxylic acids having up to 24 carbon atoms in total, such as diisopropyl adipate, diisobutyl adipate. Diisopropyl sebacate, diisopropyl maleate, or diisopropyl fumarate. Other penetration enhancing substances include phosphatidyl derivatives such as lecithin or cephalin, terpenes, amides, ketones, ureas and their derivatives, and ethers such as dimethylisosorbide and diethylene glycol monoethyl ether. Suitable penetration enhancing formulations may also include mixtures of one or more of the following: mono-or polyhydric alcohols, saturated or unsaturated C₈-C₁₈Fatty alcohol, saturated or unsaturated C₈-C₁₈Fatty acids, saturated or unsaturated fatty esters having up to 24 carbon atoms, diesters of saturated or unsaturated dicarboxylic acids having up to 24 carbon atoms in total, phosphatidyl derivatives, terpenes, amides, ketones, ureas and their derivatives, and ethers.

Adhesive materials suitable for use in transdermal delivery systems are known to those skilled in the art and include polyacrylates, silicones, polyurethanes, block polymers, styrene-butadiene copolymers, and natural and synthetic rubbers. Cellulose ethers, derivatized polyethylenes, and silicates may also be used as matrix components. Other additives such as tackifying resins or oils may be added to increase the tackiness of the matrix.

For all dosing regimens disclosed herein for compounds of formula I, the daily oral dose is preferably 0.01-200mg/kg of total body weight. For administration by injection, including intravenous, intramuscular, subcutaneous and parenteral injection, as well as using infusion techniques, the daily dose is preferably from 0.01 to 200mg/kg of total body weight. For rectal administration, the daily dose is preferably from 0.01 to 200mg/kg of total body weight. For vaginal administration, the daily dose is preferably 0.01-200mg/kg of total body weight. For topical administration, the daily dose is preferably 0.1-200mg, administered 1-4 times per day. The transdermal concentration is preferably the concentration required to maintain a daily dose of 0.01-200 mg/kg. For administration by inhalation, the daily dose is preferably from 0.01 to 10mg/kg of total body weight.

It will be appreciated by those skilled in the art that the particular method of administration will depend on a number of different factors, all of which are routinely considered when administering a treatment. It will be understood, however, that the specific dose level for any given patient will depend upon a variety of different factors including, but not limited to, the activity of the specific compound employed, the age of the patient, the weight of the patient, the general health of the patient, the sex of the patient, the diet of the patient, the time of administration, the route of administration, the rate of secretion, the combination of drugs, and the severity of the condition being treated. It will be appreciated by those skilled in the art that the optimal course of treatment, i.e. the mode of treatment and the number of daily administrations of a compound of formula I or a pharmaceutically acceptable salt thereof over a determined number of days of treatment, can be determined by those skilled in the art using routine therapeutic trials.

General preparation method

The compounds of the present invention can be prepared by using known chemical reactions and methods. However, synthesis of KDR inhibitors is aided to the reader by the following general preparation methods, and more detailed specific examples are provided in the experimental section describing the working examples.

All variable groups in these methods are as described in the general description below, if not specifically defined. It is understood that in a given structure, when having a variable group or substituent of a given symbol (i.e., R)³、R⁴、R⁶、G¹、G²、G³Or G⁴) When used more than once, each of these groups or substituents independently varies within the definitions of the symbols. As defined above, the compounds of the invention contain ring units, each of which independently may carry from 0 to 5 substituents G not defined as H¹、G³Or G⁴. In contrast, it should be noted that in the general process reaction scheme described below, G is used¹、G³Or G⁴The definition of substituent includes H to indicate G¹、G³Or G⁴The position of the substituents in the structure and can be easily drawn with precision. However, this non-standard usage is not intended to change G¹、G³Or G⁴The definition of (1). Thus, G is only for the purpose of the following general method reaction scheme¹、G³Or G⁴Except that is G¹、G³Or G⁴May be H in addition to the groups listed in the definition of (a). The final compound contains 0-5 non-hydrogen groups G¹、G³Or G⁴。

In these general methods, the variable M is equal to the group shown below

Wherein each variable group or substituent independently varies within the limits of the definitions of the above symbols.

In these general methods, the variable Q¹Is equal to the group shown below

Wherein L is N and each variable group or substituent independently varies within the limits of the definition of the symbol above.

In these general methods, the variable Q²Is equal to the group shown below

It should be recognized that the compounds of the present invention having each of the claimed optional functional groups cannot be prepared by each of the following methods. Within the scope of each process, optional substituents which are stable under the reaction conditions, or functional groups which may participate in the reaction, are used in protected form as desired, and such protecting groups are removed at appropriate steps by methods well known to those skilled in the art.

General method A: wherein X, M, and Q²As defined above, Y is-CH₂-O-、-CH₂-S-、-CH₂-NH-, -O-, -S-, or-NH-, R¹And R²Compounds of formula I-A, which together with the carbons to which they are attached form a fused 5-membered heteroaromatic ring, hal is halogen (Cl, Br, F, or I, but preferably Cl, Br or F), are conveniently prepared according to the reaction scheme shown in Process A. Thus, heterocycles of formula II wherein R is lower alkyl can be prepared by those skilled in the art according to the corresponding published methods in the reference tables. For thiophene-2, 3-dicarboxylic acid (table item 1) and pyrazole-3, 4-dicarboxylic acid (table item 10), the carboxylic acid can be converted to the methyl or ethyl ester by treatment with the corresponding alcohol and a catalytic mineral acid (typically sulfuric acid) under reflux conditions. The diester of formula II is treated with hydrazine hydrate to obtain intermediate III (see Robba, m.; Le Guen, y.bull. soc.chem. fr., 1970124317 for specific reaction conditions). Compound III is treated with a halogenating agent, such as phosphorus oxychloride, phosphorus oxybromide, phosphorus pentabromide, or phosphorus pentachloride, to produce dihalogenated intermediate IV. The dichloro or dibromo intermediate can be converted to the difluoro intermediate (if desired) by reaction with hydrogen fluoride. By using an iodine reagent such as potassium iodide or tetrabutylammonium iodide in the subsequent step, an iodine intermediate is formed in the reaction mixture without separation into pure substances. Dihalo-intermediate IV is treated with a nucleophile of formula V in refluxing alcohol or other suitable solvent such as Tetrahydrofuran (THF), Dimethoxyethane (DME), Dimethylformamide (DMF), dimethyl sulfoxide (DMSO), etc., to obtain an intermediate of formula VI. Such condensation may also be carried out without the use of solvents in the meltIn the state and with acids such as HCl or bases such as triethylamine or 1, 8-diazabicyclo [5.4.0 ]]Undec-7-ene (DBU) catalysis. Reacting a compound of formula VI with a compound of formula VII: in a suitable aprotic solvent, e.g. DMSO, DMF, or without a solvent, usually a basic catalyst such as DBU or CsCO is used₄Or a crown ether such as 18-crown-6, typically at room temperature to reflux temperature, to form the compounds of formula I-A of the present invention. It will be appreciated that the skilled person will select the appropriate solvent, catalyst (if used) and temperature depending on the nature of the feedstock. Intermediates of formula V and VII are generally commercially available or may be conveniently prepared by methods well known to those skilled in the art. For example, with respect to where Y is-CH₂-O-, and Q²Is substituted by 2-aminocarbonyl (2-CONH)₂) Preparation of VII of substituted 4-pyridyl is described in Martin, i., et al acta.

Method A

Reference Table for preparing starting Material II

General procedureB: wherein X, M, and Q²As defined above, Y is-CH₂-O-、-CH₂-S-、-CH₂The compounds of the formula I-B-NH-, -O-, -S-, or-NH-are prepared according to process B. Isoquinolinone VIII and PBr were prepared according to methods described in the literature (Tomisawa and Wang, chem. pharm. Bull., 21, 1973, 2607, 2612)₅In the molten state to give 1, 4-dibromoisoquinoline IX. Intermediate IX is treated with a nucleophile of formula V in refluxing alcohol to produce an intermediate of formula X. Such a constrictionThe polymerization can also be carried out in the molten state without the use of solvents and can be carried out with acids such as HCl or bases such as triethylamine or 1, 8-diazabicyclo [5.4.0 ]]Undec-7-ene (DBU) catalysis. Reacting a compound of formula X with a compound of formula VII: in a suitable aprotic solvent, e.g. DMSO, DMF, or without a solvent, usually a basic catalyst such as DBU or CsCO is used₄The reaction is carried out at elevated temperature to form the compounds of formula I-B of the present invention. When Y is-CH₂-S-or-S-, the method is most useful.

Method B

General method C: m, X, R therein¹、R²M and Q²The compounds of formula I-C as defined above can be conveniently prepared according to the reaction scheme shown in Process C. In this process, m is preferably 0, R¹And R²Together with the carbons to which they are attached form a fused benzene or fused 5-membered aromatic heterocycle. Starting material XI is commercially available or can be prepared by one skilled in the art according to the methods set forth in the following reference tables. Feedstock XI is reacted with urea or ammonia, typically at elevated temperature and pressure (for ammonia), to form imide XII. The imide is reacted with aldehyde XIII in acetic acid and piperidine under reflux to form intermediate XIV. XIV is reacted with sodium borohydride in methanol or other suitable solvent to give intermediate XV following the general procedure described in i.w.elliott and y.takekoshi (j.heterocyclic chem.197613, 597). With suitable halogenating agents, e.g. POCl₃、POBr₃、PCl₅、PBr₅Or thionyl chloride to generate a halo intermediate XVI which is reacted with a nucleophile of formula V in refluxing alcohol to obtain the compounds of the invention of formulae I-C. Such condensation can also be carried out in the molten state without the use of solvents and can be carried out with acids such as HCl or bases such as triethylamine or 1, 8-diazabicyclo [5.4.0 ]]Undec-7-ene (DBU) catalysis. OrThe condensation of the reagent V with the intermediate XV can be carried out by reacting the two components with P₂O₅Heating in the molten state to form the compounds of structures I-C of the present invention. The latter method is particularly effective when X is an amine linking group.

Method C

Reference table for preparation of raw materials

General method D: wherein R is¹、R²、R⁶、M、X、Y、G³And Z is as defined above, a compound of formula I-D-1 wherein q is 0 or 1 can be prepared by the reaction scheme shown in Process D. Thus, pyridine-substituted pyridazines or pyridines (I-D-1) are functionalized to form substituted 2-aminocarbonylpyridines of the formula (I-D-2) by using formamide (XVII) in the presence of hydrogen peroxide and iron salts according to the methods described in the literature (Minisci et al, Tetrahedron, 1985, 41, 4157). When R is¹And R²The process is most preferred when the two are taken together to form a fused aromatic heterocyclic or carbocyclic ring. When Z is CH, R¹And R²When no fused aromatic ring is formed, a compound wherein Z is CCONHR may be formed⁶If formed, is separated from the desired product by chromatography.

Method D

General method E: wherein R is¹、R²、R⁶、M、X、Y、G³And Z is as defined above, q is 0 or 1, and R³Compounds of formulae I-E-1 and I-E-2, which are lower alkyl, can be prepared by the reaction scheme shown in Process E. Thus, according to the methods described in the literature (Coppa, F. et al, Tetrahedron Letters, 1992, 33(21), 3057), at S₂O₈ ^-2Acid and catalytic amount of AgNO₃The pyridine-substituted pyridazine or pyridine (I-D-1) is functionalized to the substituted 2-alkoxycarbonylpyridine of formula (I-E-1) by using oxalic acid monoalkyl ester (XVIII) in the presence of an acid monoalkyl ester (XVIII). And then forming R wherein R by hydrolyzing the ester with a base such as sodium hydroxide in methanol/water³A compound of formula I-E-1 which is H. Wherein R is⁶Compounds of formula I-E-2, independently as defined above, especially including those in which two R are⁶Compounds, neither of which is H, can be prepared from the acid (I-E-1, R) by treatment with an amine XIX in the presence of a coupling agent such as DCC (dicyclohexylcarbodiimide)³H) is conveniently prepared. When R is¹And R²The process is most preferred when the two are taken together to form a fused aromatic heterocyclic or carbocyclic ring. When Z is CH, R¹And R²Where no fused aromatic ring is formed, in a first step a compound in which Z is CCO may be formed₂R³If formed, is separated from the desired product by chromatography.

Method E

General procedure F: m, Q therein²And X is as defined above, m is an integer from 1 to 5, and R¹And R²Together with the carbon to which they are attached form a fused 5-membered aromatic heterocyclic ring of formula I-FThe compounds can be prepared by the reaction scheme shown in method F. The readily available heterocyclylcarboxylic acid starting material XX is reacted with butyllithium and then with dimethylformamide to produce the aldehyde of structure XXI. Reaction of XXI with hydrazine generates pyridazinone XXII. With suitable halogenating agents, e.g. POCl₃、POBr₃、PCl₅、PBr₅Or thionyl chloride treatment of XXII to give a halogenated intermediate, which is reacted with a nucleophile of formula V in refluxing alcohol to give an intermediate compound of formula XXIII. Such condensation can also be carried out in the molten state without the use of solvents and can be carried out with acids such as HCl or bases such as triethylamine or 1, 8-diazabicyclo [5.4.0 ]]Undec-7-ene (DBU) catalysis. Alternatively, condensation of reagent V with intermediate XXII can be carried out by reacting both components with P₂O₅Heating in a molten state is performed to produce XXII. The latter method is particularly effective when X is an amine linking group. The Reissert compound XXIII is formed and alkylated with halide XXIV following the general procedure of f.d. popp, Heterocycles, 1980, 14, 1033 to produce the intermediate of structure XXV. Followed by treatment with a base to form compounds I-F of the present invention.

Method F

General method G: m, Q therein²And X is as defined above, m is an integer from 1 to 4, and R¹And R²Compounds of formula I-G, which together with the carbon to which they are attached form a fused 5-membered aromatic heterocyclic ring, can be prepared by the reaction scheme shown in method G. Aldehyde XXI from process F can be reduced with sodium borohydride to form a hydroxy acid, which can be lactonized by methods well known to those skilled in the art, e.g., using tosyl chloride, to form lactone XXVI. Intermediate XXVI is condensed with aldehyde XIII in the presence of a base such as sodium methoxide, typically in a solvent such as methanol, at reflux to give the intermediate of structure XXVII. Reaction of XXVII with hydrazine or preferably hydrazine hydrateAt 100 ℃ and 150 ℃ to form the intermediate of structure XXVIII. Intermediate XXVIII is converted to the compounds of the invention of structures I-G using XXVIII instead of XV, following the procedure described in method C.

Method G

General method H: wherein R is¹、R²、M、X、R⁶Q and G³The compounds of formula I-H as defined above may be conveniently prepared by the reaction scheme shown in Process H. Thus, it is used in Martin, I; anvelt, J.; vares, l.; kuehn, i.; XXX is converted to XXXI by the method described in Claesson, A.Actacem.Scand.1995, 49, 230-232, or by methods D or E as described above but replacing I-D-1 with the readily available pyridine-4-carboxylate XXX. The reaction mixture is then treated with a mild reducing agent such as NaBH as described in Martin et al, supra₄The ester is reduced to leave the amide substituent unchanged to afford alcohol XXXII. The alcohol is then reacted with a base such as DBU or CsCO₄And the halopyridazine VI from Process A is heated under anhydrous conditions to form the compounds of formula I-H of this invention.

Method H

General method I: wherein R is¹、R²、M、X、R⁶Q and G³As defined above, and W is a bond or-CH₂The compounds of formula I-I of (II) can be conveniently prepared by the reaction scheme shown in Process I. This process is particularly useful when q is 1 and XXXIII is 4-chloropyridine. Alternatively, other 4-halopyridines such as 4-fluoropyridine or 4-bromopyridine may be used in the process. Using the above methodGeneral procedure for Process D or E, conversion of the readily available 4-halopyridine XXXIII to an intermediate of formula XXXIV by substituting the 4-halopyridine for I-D-1. XXXIV is reacted with potassium or sodium hydrosulfide to produce a thiol of formula XXXV. Alternatively, the alcohol function of intermediate XXXII from process H is converted to a leaving group by reaction with methanesulfonyl chloride and a suitable base such as triethylamine under low temperature conditions (which minimizes polymer formation) and the resulting intermediate is reacted with potassium or sodium hydrosulfide to produce a thiol of formula XXXVI. A thiol of the formula XXXV or XXXVI is reacted with intermediate VI from Process A and a suitable base, for example diisopropylethylamine or CsCO₄In DMF or other suitable anhydrous solvent or in the absence of solvent to form I-D-9.

Method I

General procedure J: wherein R is¹、R²M, X, W, and G³Compounds of formula I-J-1 or I-J-2, as defined above, and having a sulfoxide or sulfone within the structure, are conveniently prepared by the reaction scheme shown in Process J. Containing a thioether as substituent G by treatment with one equivalent of m-chloroperbenzoic acid in dichloromethane or chloroform (MCPBA, Synth. Commun. 26, 10, 1913-1920, 1996) or by treatment with sodium periodate in methanol/water at 0 ℃ to room temperature (J.Org. chem. 58, 25, 6996-7000, 1993)¹、G³Or G⁴A portion of the compound of the invention, or as part of Y as shown in representative structures I-I from Process I, is converted to a compound of the invention having a sulfoxide group, e.g., I-J-1. The expected by-products, consisting of the various N-oxides and sulfones I-J-2, can be removed by chromatography. The sulfone I-J-2 is prepared by reacting an additional equivalent of MCPBA or preferably potassium permanganate in acetic acid/water (Eur.J.Med.chem.Ther., 21, 1, 5-8, 1986) or in acetic acid with hydrogen peroxide (chem.heterocyclic. Compd., 15, 1085-1088,1979) and then obtaining the product. When the undesired N-oxides become the major products, they can be converted back to the desired sulfoxide or sulfone by hydrogenation with palladium on carbon catalysts in ethanol/acetic acid (Yakugaku zashi, 69, 545. sup. 548, 1949, chem. Abstr.1950, 4474).

General method K: wherein R is¹、R²M, X, and Q¹The compounds of the formulae I to K according to the invention as defined above can be prepared conveniently by the reaction scheme shown in Process K. The starting materials of structure XXXVII can be prepared by methods known in the literature by those skilled in the art. For example, wherein R¹And R²XXXVII, which together with the carbon to which they are attached form a 2, 3-substituted thiophene, furan, pyrrole, cyclopentadiene, oxazole or thiazole, can be prepared by the method described in j. Pyrazole starting materials can be prepared by reacting 2-oxo-3-pentyne-1, 5-dioic acid (j.chem.phys.1974, 60, 1597) with diazomethane. Wherein R is¹And R²The starting materials which form phenyl groups together with the carbons to which they are attached can be prepared by the method of Cymerman-Craig et al, aust.j.chem.1956, 9, 222, 225. Wherein R is¹And R²Compounds of formula XXXVII, which are lower alkyl groups, can be conveniently prepared according to the procedures described in patent CH 482415(chem. abstr.120261u, 1970). The crude diacid of formula XXXVII is then treated with hydrazine to form pyridazinone XXXVIII (see Vaughn, W.R.; Baird, S.L.J.Am.chem.Soc.1946681314 for specific reaction conditions). Treatment of pyridazinone XXXVIII with a halogenating agent such as phosphorus oxychloride gives a dichloro intermediate which is hydrolysed by aqueous treatment to give the chloropyridazine XXXIX. The chlorous acid XXXIX is treated with a nucleophile of formula V in the presence of a base such as sodium hydride in a solvent such as DMF or in the absence of a solvent. According to Tilley, j.w.; coffen d.l.schaer, b.h.; method of Lind, J.J.org.chem.1987522469 usingProtogens such as BH₃THF reduced the resulting acid XXXX. The product alcohol XXXXI is reacted with a base and optionally substituted 4-halopyridyl, optionally substituted 4-halopyrimidinyl or optionally substituted 4-halopyridazinyl (XXXXII) to obtain the compounds of formulae I-K of the present invention (for specific reaction conditions see Barlow, J.J; Block, M.H; Hudson, J.A.; Leach, A.; Longridge, J.L.; Main, B.g.; Nicholson, S.J.Org. Chem.1992575158).

Method K

General procedure L: wherein R is¹、R²M, X, and Q¹The compounds of formulae I-L of the present invention as defined above are conveniently prepared by the reaction scheme shown in Process L. The alcohol of formula XXXXI from process K is reacted with methanesulfonyl chloride in the presence of a suitable base, followed by reaction with potassium or sodium hydrosulfide to form the thiol xxxiii. This thiol is then reacted with the 4-halopyridine XXXXII from Process K in the presence of a suitable base, such as triethylamine, to give the compounds I-K of the invention. Alternatively, XXXXI is converted to a halogenated intermediate of formula xxxxxiv by methods well known to those skilled in the art, and the halide is reacted with a thiol XXXXV to form I-K. Intermediate XXXXIV can also be converted to intermediate xxxiii by treatment with KHS or NaHS. Reagents xxxxxv are commercially available, e.g., 4-mercaptopyridine, or can be prepared by one skilled in the art by method I above.

Method L

Detailed Description

Experiment:

example 1: preparation of 1- (4-chlorophenylamino) -4- (4-pyridylthio) isoquinoline

Step 1: preparation of intermediate a: 2.90g, 19.07mMol isoquinolone and 14.40g, 33.68mMol phosphorus pentabromide were melted together at 140 ℃. The melt was converted to a red liquid and after about 10 minutes the reaction mixture solidified and was cooled. The reaction mixture was crushed and poured into ice water. The resulting solid was filtered and air dried. wt.5.50g, 96% yield, mp. ═ 94-96 ℃. R_f0.66 in a 40% mixture of ethyl acetate in hexane.

Step 2: 1.00g, 3.49mMol from step 1, 4-two bromine isoquinoline (intermediate A) and 4-chloro aniline mixture at 140 degrees C melting together. The reaction mixture turned into a dark red liquid and solidified after about 10 minutes and was cooled. The reaction mixture was crushed and triturated with 50/50 methanol/THF mixture, then filtered and air dried without further purification. wt.0.75g, 64.4%, mp. ═ 260 and 263 ℃. R_f0.58 in a 40% mixture of ethyl acetate in hexane.

And step 3: 0.05g, 0.1498mMol 1- (4-chloro aniline) -4-bromine isoquinoline and 0.02g, 0.18mMol 4-mercapto pyridine were mixed,and melted at 140 c for about 10 minutes. The resulting reaction mixture was purified on 1000 micron preparation plates using a mixture of 5% methanol in hexane as solvent. wt.0.0103g, 19% yield, mp. ═ 192-. R_f0.50 in a 40% mixture of ethyl acetate in hexane.

Example 2: preparation of 1- (indan-5-ylamino) -4- (4-pyridylthio) isoquinoline

The title compound was prepared using the method used for preparation example 1, substituting 5-aminoindan for 4-chloroaniline in step 2. Melting point 100 ℃ and 103 ℃, TLC R_f0.40 (40% mixture of ethyl acetate in hexane).

Example 3: preparation of 1- (benzothiazol-6-ylamino) -4- (4-pyridylthio) isoquinoline

The title compound was prepared using the procedure used for the preparation of example 1, substituting 6-aminobenzothiazole for 4-chloroaniline in step 2.

TLC R_f0.36 (5% methanol/dichloromethane); MS 387

Example 4: preparation of 1- (4-chlorophenylamino) -4- (4-pyridylmethyl) isoquinoline

Step 1: high phthalimide (770mg, 4.78mmol) and 4-picolyl were addedA mixture of aldehyde (0.469mL, 4.78mmol) and piperidine (0.5mL) in acetic acid (25mL) was heated at reflux for 1 hour. The resulting solution was cooled to room temperature. The solid product was removed by filtration, washed with water (4X 10mL) and dried in vacuo to yield 920mg (3.67mmol, 77% yield) of a mixture of Z and E isomers of the above compound.¹H-NMR(DMSO-d₆) Complex proton signals are shown in the aromatic region due to the presence of both the E and Z isomers. MS ES 251(M + H)⁺，252(M+2H)⁺。

Step 2: to a suspension of starting material (1.70g, 6.8mmol) in methanol (250mL) at 0 deg.C was slowly added sodium borohydride (3.0g, 79 mmol). The mixture was warmed to room temperature and stirring was continued for 1 hour. The reaction was quenched with water (10mL) and stirred for 10 min. The resulting mixture was concentrated to remove the solvent. Ice water (100mL) was added to the residue, and the pH was adjusted to 2 with 2N hydrochloric acid. After stirring for 10 minutes, 2N NaOH was added until the pH of the solution was about 11. The resulting solution was extracted with dichloromethane (4X 100 mL). The combined organic layers were collected, dried over magnesium sulfate and concentrated. The residue was purified by column chromatography (1: 10v/v methanol-dichloromethane) to give 400mg of the title compound as a solid (1.70mmol, 25% yield).

¹H-NMR(MeOH-d₄)8.33 to 8.39(m, 4H), 7.50-7.68(m, 3H), 7.30-7.31(m, 2H), 7.14(s, 1H), 4.15(s, 2H); MS ES 237(M + H) +, 238(M + 2H); TLC (1: 10v/v methanol-dichloromethane) R_f＝0.40。

And step 3: 4-chloroaniline (178mg, 1.40mmol), phosphorus pentoxide (396mg, 1.40mmol) and triethylamine hydrochloride (193mg, 1.40mmol) were heated under argon at 200 ℃ with stirring for 1.5 hours or until a homogeneous melt was formed. To the melt was added the starting material (82mg, 0.35 mmol). The reaction mixture was stirred at 200 ℃ for 2 hours. The resulting solid black material was cooled to 100 ℃. Methanol (5mL) and water (10mL) were added and the reaction mixture was sonicated until the black material was dissolved. Dichloromethane (40mL) was added and concentrated aqueous ammonia (2mL) was added to adjust the mixture to pH 10. The organic layer was separated and the aqueous layer was extracted with dichloromethane (3X 20 mL). The combined organic layers were dried over magnesium sulfate, filtered, and concentrated. Purification by preparative TLC plate (1: 10v/v methanol-dichloromethane) gave 26mg (0.08mmol, 22% yield) of the title compound as a yellow solid.

¹H-NMR(MeOH-d₄)8.37(d，J＝7.8Hz，3H)，7.86(s，1H)，7.55-7.77(m，5H)，7.27-7.33(m，4H)，4.31(s，2H)；MS ES 346(M+H)⁺；

TLC (1: 10v/v methanol-dichloromethane) R_f＝0.45。

Example 5: preparation of 1- (benzothiazol-6-ylamino) -4- (4-pyridylmethyl) isoquinoline

The title compound was prepared using the procedure used for the preparation of example 4 substituting 6-aminobenzothiazole for 4-chloroaniline in step 3.

¹H-NMR(MeOH-d₄)9.08(s，1H)，8.37-8.59(m，4H)，7.79-8.01(m，2H)，7.60-7.78(m，4H)，7.30(d，2H)，4.34(s，2H)；MS ES 369(M+H)⁺；

TLC (1: 4v/v hexane-ethyl acetate) R_f＝0.20。

Example 6: preparation of 1- (indan-5-ylamino) -4- (4-pyridylmethyl) isoquinoline

The title compound was prepared using the procedure used for the preparation of example 4 substituting 5-aminoindan for 4-chloroaniline in step 3.

¹H-NMR(MeOH-d₄)8.35(m，3H)，7.46-7.77(m，5H)，7.15-7.27(m，4H)，4.26(s，2H)，2.87-2.90(m，4H)，2.05-2.10(m，2H)；MS ES 352(M+H)⁺；

TLC (1: 4v/v hexane-ethyl acetate) R_f＝0.25。

Example 7: preparation of 1- (3-fluoro-4-methylphenylamino) -4- (4-pyridylmethyl) isoquinoline

The title compound was prepared using the procedure used for the preparation of example 4 substituting 3-fluoro-4-methylaniline for 4-chloroaniline in step 3.

¹H-NMR(MeOH-d₄)8.34(d，3H)，7.87(s，1H)，7.54-7.69(m，4H)，7.10-7.31(m，4H)，2.22(s，3H)；MS ES 344(M+2H)⁺；

TLC (1: 4v/v hexane-ethyl acetate) R_f＝0.20。

Example 8: preparation of 4- (4-chlorophenylamino) -7- (4-pyridylmethoxy) thieno [2, 3-d ] pyridazine

Step 1: a dry 2L three-necked round-bottomed flask was fitted with a mechanical stirrer and an addition funnel. To the flask was added 2-thiophenecarboxylic acid (25g, 195mmol) in anhydrous THF (500mL) under argon. The mixture was cooled to-78 ℃ with a dry ice-isopropanol bath and stirred for 30 minutes. A solution of n-butyllithium in hexane (2.5M, 1.72mL) was added dropwise over 30 minutes. The reaction was maintained at-78 ℃ for 1 hour with stirring and then placed under an atmosphere of dry carbon dioxide. With the addition of carbon dioxide, the reaction thickened. The reaction was held at-78 ℃ for 1 hour and then warmed to-10 ℃. The reaction was quenched with 2N HCl (213mL) and allowed to reach room temperature. The layers were separated and the aqueous layer was extracted with ethyl acetate (3X 200 mL). The organic layers were combined, dried (sodium sulfate) and concentrated by rotary evaporation. The brown solid was crystallized from hot isopropanol and dried in vacuo overnight. The desired thiophene-2, 3-dicarboxylic acid (27.3g, 159 mmol; 82% yield) was obtained;

¹H NMR(DMSO-d₆)7.69(d，J＝1.5，1)，7.38(d，J＝4.8，1)；ES MS(M+H)⁺173; TLC (chloroform-MeOH-water, 6: 4: 1); r_f＝0.74。

Step 1A: alternatively, the same product was obtained using 3-thiophenecarboxylic acid instead of 2-thiophenecarboxylic acid in step 1.

Step 2: A1L round bottom flask was fitted with a stir bar and reflux condenser. The flask was charged with the product of step 1 (62g, 360mmol) in methanol (500mL) containing a catalytic amount of sulfuric acid (. about.5 mL). The reaction was heated to reflux and stirred for 24 hours. The reaction was cooled to room temperature and concentrated by rotary evaporation. The brown mixture is purified by chromatography on silica gel (hexane-ethyl acetate gradient 80: 20-60: 40). The desired thiophene-2, 3-dicarboxylic acid dimethyl ester (21.2g, 106 mmol; yield 31%) was obtained;

¹H NMR(DMSO-d₆)7.93(d，J＝4.8，1)，7.35(d，J＝4.8，1)，3.8(d，J＝1，6)；ES MS(M+H)⁺＝201；

TLC (Hexane-EtOAc, 70: 30); r_f＝0.48。

And step 3: a250 mL round bottom flask was equipped with a stir bar and reflux condenser. To the flask were added the product of step 2 (16g, 80mmol), hydrazine hydrate (6.6mL, 213mmol), and ethanol (77mL), and refluxed for 2.5 hours. The reaction was cooled to room temperature and concentrated by rotary evaporation. Water (50mL) was added and the filtrate was separated from insoluble solids. The aqueous layer was concentrated by rotary evaporation to give a pale yellow solid. The solid was dried in a vacuum oven at 50 ℃ overnight. The desired thieno [2, 3-d ] pyridazine-4, 7-dione (12g, 71 mmol; yield 89%) was obtained;

¹H NMR(DMSO-d₆)7.85(d，J＝5.1，1)，7.42(d，J＝5.1，1)；ES MS(M+H)⁺＝169；

TLC (dichloromethane-methanol, 60: 40); r_f＝0.58。

And 4, step 4: preparation of intermediate B: a250 mL round bottom flask was equipped with a stir bar and reflux condenser. To the flask were added the product of step 3 (2.5g, 14.8mmol), phosphorus oxychloride (45mL, 481mmol), and pyridine (4.57mL, 55mmol), and refluxed for 2.5 hours. The reaction was cooled to room temperature and poured onto ice. The mixture was separated and the aqueous layer was extracted with chloroform (4X 75 mL). The organic layers were combined, dried (sodium sulfate), and concentrated by rotary evaporation to give a dark yellow solid. The desired 4, 7-dichlorothieno [2, 3-d ] pyridazine (intermediate B; 1.5g, 7.3 mmol; yield 49%); the melting point is 260-263 ℃;

¹H NMR(DMSO-d₆)8.55(d，J＝5.7，1)，7.80(d，J＝5.7，1)；ES MS(M+H)⁺＝206；

TLC (Hexane-EtOAc, 70: 30); r_f0.56. See also Robba, m.; bull.soc.chim.fr.; 1967, 4220-4235.

And 5: a250 mL round bottom flask was equipped with a stir bar and reflux condenser. To the flask was added the product of step 4 (7.65g, 37.3mmol), 4-chloroaniline (4.76g, 37.3mmol) in ethanol (75 mL). The mixture was refluxed for 3 hours. An orange solid precipitated from the reaction after 3 hours. The reaction was cooled to room temperature and the solid was collected by filtration and washed with hexanes. The desired 7-chloro-4- (4-chlorophenylamino) thieno [2, 3-d ] is obtained]Pyridazine (6.5g, 21.9 mmol; yield 60%); the melting point is 139-142 ℃; ES MS (M + H)⁺297; TLC (Hexane-EtOAc 60: 40); r_f＝0.48。

Step 6: a150 mL round bottom flask was equipped with a stir bar and reflux condenser. To the flask was added the product of step 5 (0.33g, 1.1mmol), 4-pyridylcarbinol (1.2g, 11.2mmol) in DBU (2.5mL, 16.7mmol) and the mixture was heated at 125 ℃ for 24 h. Ethyl acetate (10mL) was added to the still hot reaction, which was then reactedIt should be poured into water (10 mL). The layers were separated and the aqueous layer was extracted with ethyl acetate (3X 10 mL). The organic phases were combined, dried (magnesium sulfate) and concentrated by rotary evaporation. The resulting mixture was purified by chromatography on silica gel (dichloromethane-methanol-acetone, 90: 5) to obtain a pale yellow solid. The desired title compound was obtained (0.03g, 0.08 mmol; yield 7.3%); melting point 203-; ES MS (M + H)⁺369; TLC (dichloromethane-methanol-acetone, 95: 2.5); r_f＝0.56。

Example 9: preparation of 4- (4-chlorophenylamino) -7- (4-pyridylmethoxy) furo [2, 3-d ] pyridazine

Step 1: to a dry 3L three-necked flask equipped with an addition funnel, argon inlet, and mechanical stirrer was added n-butyllithium (2.5M in hexane, 196mL, 491 mmol). The mixture was diluted with anhydrous THF (500mL) and cooled to-78 ℃. A solution of 3-furancarboxylic acid (25g, 223mmol) in THF (500mL) was added dropwise. The mixture was stirred for 1.5 hours, and anhydrous carbon dioxide was passed through the reaction mixture for 1 hour. After gradually warming to-10 ℃, the resulting thick white slurry was treated with hydrochloric acid (2N, 446 mL). The layers were separated and the aqueous layer was extracted with ethyl acetate (3X 300 mL). The combined organic layers were dried (sodium sulfate), filtered, and concentrated to give crude furan-2, 3-dicarboxylic acid as an orange solid (44g) which was used without further purification.

¹H NMR(300MHz，d₆-acetone) δ 7.06(d, J ═ 1.7, 1), 7.97(d, J ═ 1.7, 1), 10.7(bs, 2H); TLC (CHCl)₃/MeOH/H₂O 6∶4∶1)R_f＝0.56.

Step 2: a dry 500mL round bottom flask was fitted with a stir bar and argon inlet. To the flask was added the crude diacid product prepared in step 1 (44g) dissolved in methanol (250 mL). Chlorotrimethylsilane (80mL, 630mmol) was added portionwise to the reaction mixture. After stirring at room temperature for 15.5 h, the solution was concentrated to an oil and silica (5g) was added. The mixture was suspended in methanol (100mL) and the volatiles were removed. The operation of suspending in methanol (100mL) and removing the volatile material was repeated 2 more times. The residue was applied directly to the top of a flash column, eluting with hexane/ethyl acetate 60: 40, to give dimethyl furan-2, 3-dicarboxylate as an orange oil (38g, 93% yield for combined step 1 and step 2).

¹H NMR(300MHz，CDCl₃) δ 3.81(s, 3), 3.86(s, 3), 6.71(d, J ═ 2.8, 1), 7.46(d, J ═ 2.8, 1); TLC (Hexane/EtOAc 60: 40) R_f＝0.46.

And step 3: to a 500mL round bottom flask equipped with an argon inlet, reflux condenser, and stir bar was added dimethyl furan-2, 3-dicarboxylate (44g, 236mmol) dissolved in ethanol (250 mL). To this solution was added hydrazine hydrate (55% N)₂H₄40mL, 3.0mmol) and the reaction mixture is heated to reflux. A yellow solid precipitated slowly over 5.5 hours and the mixture was cooled to room temperature. Volatiles were removed under reduced pressure to give a yellow paste which was suspended in water and filtered. The yellow solid was washed with water and transferred to a 500mL round bottom flask equipped with an argon inlet, reflux condenser, and stir bar. The solid was suspended in hydrochloric acid (2N, 200mL) and the mixture was heated to reflux. After heating for 4 hours, the orange slurry was cooled to room temperature and filtered. By usingThe solid was washed well with water and dried under vacuum to obtain 4, 7-dioxo [2, 3-d ]]Furopyridazine as an orange solid (21.5g, 60%).

¹H NMR(300MHz，d₆-DMSO)δ7.00(d，J＝2.1，1)，8.19(d，J＝2.1，1H)，11.7(bs，2H).

And 4, step 4: preparation of intermediate C: A1L round bottom flask was fitted with a reflux condenser, stir bar, and argon inlet. Step 3 furan (15.5g, 102mmol) was added to a mixture of phosphorus oxychloride (300mL) and pyridine (30mL) and the resulting orange suspension was heated to reflux. After heating the reaction mixture for 4 hours, the volatiles were removed by rotary evaporation. The residue was poured onto ice and the aqueous mixture was extracted with chloroform (4X 250 mL). The combined organic layers were washed with brine, dried (magnesium sulfate), and concentrated to give 4, 7-dichloro [2, 3-d ]]Furopyridazine (intermediate C, 11.3g, 59%) was used as an orange-red solid without further purification. TLC (Hexane/EtOAc) R_f＝0.352；

¹H NMR(300MHz，d₆-DMSO)δ7.40(d，J＝2.0，1)，8.63(d，J＝2.0，1).

And 5: to a 100mL round bottom flask equipped with a stir bar, argon inlet, and reflux condenser was added the product of step 4 (1.50g, 7.98mmol) dissolved in ethanol (40 mL). Chloroaniline (1.02g, 7.98mmol) was added to the mixture and the resulting suspension was heated to reflux. After heating for 4 hours, the mixture was concentrated by rotary evaporation. The crude orange solid was applied to the top of a flash column and eluted with methylene chloride/methanol 97: 3 to give a mixture of 4-chloro-7- [ N- (4-chlorophenyl) amino ] [2, 3-d ] furopyridazine and 7-chloro-4- [ N- (4-chlorophenyl) amino ] [2, 3-d ] furopyridazine as a yellow powder (1.2g, 55%).

TLC(CH₂Cl₂/MeOH 97∶3)；R_f＝0.7；¹H NMR(300MHz，d₆-DMSO) δ major isomer (a)7.40(d, J ═ 8.9, 2), 7.45(d, J ═ 2.0, 1), 7.87(d, J ═ 9.2, 2), 8.34(d, J ═ 2.0, 1)9.62(s, 1); minor isomer (B)7.28(d, J ═ 2.0, 1), 7.40(d, J ═ 8.9, 2), 7.87(d, J ═ 9.2, 2), 8.48(d, J ═ 2.1, 1), 9.88(s, 1).

Step 6: a25 mL round bottom flask was equipped with an argon inlet, stir bar, and reflux condenser. The product of step 5 (400mg, 1.4mmol) was reacted with 4-pyridylcarbinol (782mg, 7.17mmol) and 1, 8-diazabicyclo [5.4.0 ]]Undec-7-ene (2.5mL, 16.7mmol) was mixed and the slurry was heated to 125 ℃. After stirring for 24 hours, the reaction was cooled, applied directly to the top of the flash column and eluted with dichloromethane/methanol 95: 5. The resulting yellow oil was repurified by chromatography under the same conditions to afford the title compound as part of a three component mixture. HPLC separation (C)₁₈Column, CH₃CN/H₂O10: 90-100: 0 gradient) to give the title compound as an off-white solid (13.7mg, 3%).

TLC(CH₂Cl₂/MeOH 95∶5)＝0.19；MP198℃；¹H NMR(300MHz，CDCl₃)δ5.60(s，2)，6.6(d，J＝2.1，1)，7.18-7.20(m，2)，7.35-7.43(m，6)，7.66(d，J＝2.1，1)8.54(d，J＝5.6，2).

Steps 5A and 6A: alternatively, following the procedure of step 5, but substituting the dichloro intermediate with the dibromo intermediateUsing 4, 7-dibromo [2, 3-d ]]Furopyridazine (hereinafter intermediate G) to prepare the title compound. By the addition of carbon dioxide in CsCO₄Rather than 1, 8-diazabicyclo [5.4.0 ]]Step 6A was performed by melting the two components in the presence of undec-7-ene. The crude product was purified as described above.

Intermediates D to G: preparation of other bicyclic 4, 5-fused-3, 6-dihalopyridazines

The general procedure of example 9, steps 2-4 was followed, substituting the appropriate heterocyclic dicarboxylic acid for furan-2, 3-dicarboxylic acid to afford the substituted dichloropyridazines D-G shown in the following Table. Dibromofuropyridazine G was prepared using the procedure of example 9, steps 2-3, followed by step 4' as follows: to 0.50g (3.287mmol) of the product of step 3 was added 2.83g (6.57mmol) of phosphorus pentabromide. The mixture was heated to 125 ℃. The reaction mixture melted at about 115 ℃ and then solidified again before reaching 125 ℃. The reaction mixture was cooled and the solid residue was crushed and poured into ice water. The resulting solid was then filtered off and dried in vacuo. wt. -, 0.75g (82% yield). As shown in the given reference, dichloropyridazine is a known substance in several cases. All of these dihaloheterocycles can be used to prepare the compounds of the present invention.

Intermediate H: preparation of (2-methylaminocarbonyl-4-pyridyl) methanol

Step 1: a stirred solution of ethyl isonicotinate (250mL, 1.64mol) and concentrated sulfuric acid (92mL, 1.64mol) in N-methylformamide (2.0L) was cooled to 6 ℃ with an ice bath. Iron (II) sulfate heptahydrate (22.8g, 0.0812mol, ground with a mortar and pestle) was added, followed by dropwise addition of 30% aqueous hydrogen peroxide (56mL, 0.492 mol). The addition of iron (II) sulfate and hydrogen peroxide was repeated 4 more times while maintaining the reaction temperature below 22 ℃. After stirring the reaction mixture for 30 minutes, sodium citrate solution (2L, 1M) was added (pH of the resulting mixture was about 5). The mixture was extracted with dichloromethane (1L, 2X 500 mL). The combined organic extracts were washed with water (2X 500mL), 5% aqueous sodium bicarbonate (3X 100mL), and brine (500 mL). The resulting organic solution was then dried over sodium sulfate, filtered and concentrated in vacuo to afford a solid. The crude solid was triturated with hexane, filtered, washed with hexane and dried under vacuum to give 270.35g (79.2%) of a yellow solid as a powder.

¹H NMR(DMSO-d₆，300MHz)：δ8.9(d，1H)，8.3(m，1H)，8.0(dd，1H)，4.4(q，2H)，2.8(d，3H)，1.3(t，3H).

Step 2: to a mechanically stirred slurry of the product of step 1 (51.60g, 0.248mol) in ethanol (1.3L) was added sodium borohydride (18.7g, 0.495 mol). The reaction mixture was stirred at room temperature for 18 hours. The resulting aqueous solution was carefully quenched with saturated aqueous ammonium chloride (2L). During quenching, gas evolution was observed. The resulting mixture was basified with concentrated ammonium hydroxide solution (200ml) to pH 9. Then extracted with ethyl acetate (8X 400 mL). The combined organic layers were dried (magnesium sulfate), filtered, and concentrated in vacuo to afford intermediate H as a clear pale yellow oil (36.6g, 89% yield).

¹H NMR(DMSO-d₆，300MHz)：δ8.74(q，1H)，8.53(dd，1H)，7.99(m，1H)，7.48(m，1H)，5.53(t，1H)，4.60(d，2H)，2.81(d，3H)；MS m/z 167[M+H]⁺.

Intermediates I to N: general procedure for the preparation of [2- (N-substituted) aminocarbonyl-4-pyridinyl ] methanol intermediates

To a solution of amine 2(3 equivalents) in benzene was added trimethylaluminum (3 equivalents) at 0 ℃. Evolution of gas was observed, then warmed to room temperature and stirred for 1 hour (Lipton, m.f. et al, org. synth. col. vol.6, 1988, 492 or Levin, j.i. et al, synth. comm., 1982, 12, 989). Known methanol 1(1 equivalent, Hadri, A.E.; Leclerc, G.Heterocyclic Chem, 1993, 30, 631) was added to the aluminium reagent and the mixture was heated to reflux for 1 hour. The reaction was quenched with water and concentrated. The crude product was purified by column chromatography on silica gel (20/1 ethyl acetate/methanol) to give the title compound 3. The final product was generally confirmed by LC/MS and NMR spectroscopy.

Dichloromethane was used as solvent instead of benzene.

Example 10: preparation of 4- (4-chlorophenylamino) -7- (2-aminocarbonyl-4-pyridylmethoxy) thieno [2, 3-d ] pyridazine

A25 mL three-necked round-bottomed flask was equipped with a stir bar and thermometer. The flask was charged with the product of example 8 (0.475g, 1.29 mm)ol), iron sulfate heptahydrate (0.179g, 0.64mmol), formamide (11.15mL, 281mmol), and concentrated sulfuric acid (0.14 mL). The mixture was stirred at room temperature for 30 minutes, and then H was added dropwise to the mixture₂O₂(0.2mL, 6.44 mmol). The reaction was stirred at room temperature for an additional 1 hour and then heated to 55 ℃ over 30 minutes. The reaction was held at this temperature for 3 hours and then cooled to room temperature. Aqueous sodium citrate (0.27M, 1mL) was added to the reaction, the layers were separated, and the aqueous layer was extracted with ethyl acetate (4X 5 mL). The organic layers were combined, dried (magnesium sulfate), and concentrated by rotary evaporation. The resulting solid was placed in hot acetone and separated from any remaining solid by filtration. The filtrate was then concentrated by rotary evaporation and the resulting residue was taken up in hot methanol and a white solid was collected by filtration. The desired compound (0.14g, 0.034 mmol; yield 2.7%); melting point 233 deg.C; ES MS (M + H)⁺412; TLC (dichloromethane-methanol-acetone, 95: 2.5); r_f＝0.20。

Example 11: preparation of 4- (4-chlorophenylamino) -7- (2-methylaminocarbonyl-4-pyridylmethoxy) thieno [2, 3-d ] pyridazine

The title compound was prepared using the procedure used for the preparation of example 10, substituting methylformamide for formamide:

1H NMR(DMSO-d6)8.80(d，1)，8.62(d，1)，8.31(d，1)，8.09(d，2)，7.86(d，2)，7.65(d，1)，7.35(d，2)，5.74(s，2)，2.84(d，3)；ES MS(M+H)+＝426(ES)；R_f(95/2.5/2.5 DCM/MeOH/acetone) ═ 0.469.

Example 12: preparation of 1- (4-chlorophenylamino) -4- (2-aminocarbonyl-4-pyridylmethyl) isoquinoline

The title compound was prepared using the procedure used for the preparation of example 10 substituting the product of example 4 for the product of example 8. The crude product was purified by preparative TLC plates (1: 4v/v hexane-ethyl acetate, 19% yield) to afford the title compound as a yellow solid.

¹H-NMR(MeOH-d₄)8.42(d，1H)，8.34(d，1H)，7.94(s，1H)，7.88(s，1H)，7.55 to7.76(m，5H)，7.26 to 7.36(m，3H)，4.34(s，2H)；MS ES 389(M+H)⁺；

TLC (1: 4v/v hexane-ethyl acetate) R_f＝0.44。

Example 13: preparation of 1- (4-chlorophenylamino) -4- (2-methylaminocarbonyl-4-pyridylmethyl) isoquinoline

The title compound was prepared using the procedure used for the preparation of example 11 substituting the product of example 4 for the product of example 8. The crude product was purified by column chromatography (2: 3v/v hexane-ethyl acetate, 20% yield) to afford the title compound as a yellow solid.

¹H-NMR(MeOH-d₄)8.42(d，1H)，8.33(d，1H)，7.88(d，2H)，7.55 to 7.77(m，5H)，7.28 to 7.36(m，3H)，4.34(s，2H)，2.89(s，3H)；MS ES 403(M+H)⁺；

TLC (2: 3v/v hexane-ethyl acetate) R_f＝0.30。

Examples 14 and 15: preparation of 4- (4-chlorophenylamino) -7- (2-methylaminocarbonyl-4-pyridylmethoxy) furo [2, 3-d ] pyridazine and 4- (4-chlorophenylamino) -2-methylaminocarbonyl-7- (2-methylaminocarbonyl-4-pyridylmethoxy) furo [2, 3-d ] pyridazine

To a suspension of the final product of example 9 (19.20g, 54.4mmol) in N-methylformamide (200mL) and distilled water (20mL) at room temperature was added concentrated sulfuric acid (2.9mL, 54.4mmol) dropwise. The mixture was stirred until it became a clear solution. FeSO is added to the solution in one portion₄。7H₂O (1.51g, 5.43mmol) and then hydroxylamine-O-sulfonic acid (HOSA, 1.84g, 16.3mmol) was added. FeSO was added at 10 minute intervals₄.7H₂The operations of O and HOSA were repeated 11 times. HPLC analysis indicated that most of the starting material had been consumed. The reaction mixture was cooled with an ice bath. Sodium citrate solution (600mL, 1M, 600mmol) was added with vigorous stirring. The resulting suspension was stirred vigorously for a further 10 minutes. The solid was collected by filtration, washed with water (3X 100mL), and dried under vacuum at 50 ℃ for 16 h. The crude product (21g) was purified by filtration through a pad of silica gel eluting with 5% methanol in dichloromethane. 3.7g of the product obtained are in CH₃Recrystallization from CN (125mL, boiling 1.5 hours). The solid was collected by filtration and washed with CH₃CN (2X 15mL) and dried under vacuum at 50 ℃ for 16 h. The final product (4- (4-chlorophenylamino) -7- (2-methylaminocarbonyl-4-pyridylmethoxy) furo [2, 3-d]Pyridazine) was a pale yellow solid (3.38g, 15.2%). Melting point 223-.

The major by-products were separated by filtration through a pad of silica gel as described above. By passing¹H NMR, 2D NMR, elemental analysis, and MS to confirm the by-product 4- (4-chlorophenylamino) -2-methylaminocarbonyl-7- (2-methylaminocarbonyl-4-pyridylmethoxy) furo [2, 3-D]Structure of pyridazine.

¹H NMR(DMSO-d₆，300MHz)：δ9.32(brs，1H)，8.93(q，1H)，8.79(q，1H)，8.63(dd，1H)，8.12(m，1H)，7.91(m，3H)，7.70(dd，1H)，7.35(m，2H)，5.76(br s，2H)，2.81(d，6H).MS m/z 467[M+H]⁺.

Example 14A: preparation of 4- (4-chlorophenylamino) -7- (2-methylaminocarbonyl-4-pyridylmethoxy) furo [2, 3-d ] pyridazine-method 2

To a mixture of intermediate from example 9, step 5 (10.0g, 35.7mmol), intermediate H (12.4g, 74.6mmol), and 18-crown-6 (0.42g, 1.59mmol) in toluene (100mL) was added KOH powder (4.4g, 85%, 66.7mmol) in one portion at room temperature. The reaction mixture was heated to 85 ± 2 ℃ with vigorous stirring. The reaction mixture was stirred vigorously at this temperature overnight. It was then cooled to room temperature, the toluene solution decanted, and water (100mL) added to the gummy residue. The resulting mixture was stirred vigorously until it became a free-flowing suspension. The solid was collected by filtration, washed with water (2X 10mL), and dried under vacuum at 45 ℃ for 16 h. The yellow/brown solid was suspended in acetonitrile (70mL) and the suspension was stirred at reflux for 2 hours. After cooling to room temperature, the solid was collected by filtration, washed with a small amount of acetonitrile and dried under vacuum at 45 ℃ overnight. The product was isolated in 46% yield (6.73g) as a pale yellow solid.

Example 16: preparation of 4- (4-chlorophenylamino) -7- (2-aminocarbonyl-4-pyridylmethoxy) furo [2, 3-d ] pyridazine

The title compound was prepared using the procedure used for the preparation of example 14, but substituting formamide for N-methylformamide. The reaction was carried out with 500mg of the end product of example 9 and the proportional amounts of solvent and reagents.The crude product was purified by HPLC on a 75 x 30mmC18 column eluting with a linear gradient of 10-100% acetonitrile in water containing 0.1% trifluoroacetic acid at a rate of 10 ml/min for 10 minutes to yield 18mg of the title compound as a yellow solid: HPLC (50X 4.6mm YMCC18 column, linear gradient eluted with a mixture of 10-100% acetonitrile in water containing 0.1% trifluoroacetic acid at a rate of 3 ml/min for 5 min, UV detection at 254 nm) for 2.35 min, peak; MS ES 396(M + H)⁺。

Example 17: preparation of 4- (4-chlorophenylamino) -7- (benzothiazol-6-ylamino) thieno [2, 3-d ] pyridazine

To the dichloride from step 4 of example 8 (1.00g, 4.90mmol) was added p-chloroaniline (622mg, 4.90mmol) and absolute ethanol (10.0 mL). The mixture was refluxed at 95 ℃ for 2 hours and then cooled to room temperature. The yellow precipitate (2) formed is filtered off, successively using isopropanol, 4.0N KOH and H₂O, and hexane. The filtrate (2) was then mixed with 6-aminobenzothiazole (883mg, 5.88mmol) in 10mL of n-butanol and heated at 150 ℃ overnight. The reaction was cooled to room temperature and then the solvent was removed by rotary evaporation. The residue was treated with 4.0N KOH solution, extracted with dichloromethane (50mL), dried (magnesium sulfate), and the solvent was evaporated. The crude product was purified by flash chromatography on silica gel using 95% dichloromethane/methanol as eluent. The structure of the pure title compound was confirmed by LC/MS and NMR: TLC (30% ethyl acetate/hexane) R_f(3)＝0.20；

¹H NMR(DMSO)δ7.2(dd，3H)，7.38(dd，3H)，7.65(d，1H)，8.0(d，1H)，8.45(d，1H)，8.8(s，1H)；LC/MS m/z 410rt＝4.21min.

Example 18: preparation of 4- (indan-5-ylamino) -7- (benzothiazol-6-ylamino) thieno [2, 3-d ] pyridazine

The title compound was prepared using the procedure used for the preparation of example 17, substituting 5-aminoindan for 4-chloroaniline. The crude product was purified by flash column chromatography on silica eluting with 30% ethyl acetate/hexane. The structure of the pure title compound was confirmed by LC/MS and NMR: TLC (30% ethyl acetate/hexane) R_f(3)＝0.20；

(3)¹H NMR(DMSO)δ2.0(m，2H)，2.85(m，4H)，7.18(d，1H)，7.8(d，1H)，7.95(d，1H)，8.10(d，1H)，8.18(d，1H)，8.7(d，2H)，9.1(d，2H)，LC/MS m/z 414rt＝4.43min.

Example 19: preparation of 4- (5-Bromomindolin-1-yl) -7- (4-pyridylmethoxy) furo [2, 3-d ] pyridazine

4, 7-dichloro [2, 3-d ] from step 4 of example 9]Furopyridazine (95mg, 0.50mmol) and 5-bromoindoline (100mg, 0.50mmol) were refluxed at 95 ℃ for 2 hours in 60mL of anhydrous ethanol. The reaction mixture was cooled to room temperature, and the formed precipitate was filtered off, washed with isopropanol, 4.0N KOH, water, and hexane, and then dried. The purity of this intermediate was about 95% (rt ═ 4.72, (M + H)⁺350) And used in the next step without further purification. 4-Pyridylcarbinol (28mg, 0.26mmol) and sodium hydride (60%, 50mg, 1.25mmol) were stirred in 20mL of anhydrous tetrahydrofuran at 0 ℃ under argon for 20 minutes, then 44mg of the above intermediate (0.13mmol) was added. The reaction was stirred at 0 ℃ for 2 hours, andthe temperature was raised to room temperature. The mixture was stirred for a further 12 hours and the solvent was evaporated under reduced pressure. The obtained solid was dissolved in 50mL of dichloromethane and washed with potassium carbonate solution and water. The organic layer was separated, dried (magnesium sulfate), and evaporated under reduced pressure. Preparative TLC (R) on silica gel_f0.3) and eluted with dichloromethane/methanol (95: 5). The structure of the pure title compound was confirmed by LC/MS and NMR:

¹H NMR(CDCl₃)δ3.20(m，2H)，4.30～4.50(m，2H)，5.60(s，2H)，6.9～8.0(m，7H)8.60(m，2H)；LC/MS(M+H)⁺423rt＝4.49min.

example 20: preparation of 4- (4-methoxyphenylamino) -7- (2-methylaminocarbonyl-4-pyridylmethoxy) furo [2, 3-d ] pyridazine

To 4, 7-dichloro [2, 3-d ] from step 4 of example 9]Furopyridazine (400mg, 2.12mmol, 1 equiv.) and p-methoxyaniline (p-MeOC)₆H₄NH₂) (260mg, 2.12mmol, 1 equiv.) to a suspension in DME (5mL) was added water (1 mL). The resulting solution was heated at 50 ℃ for 48 hours. After cooling to room temperature, the brown precipitate was removed by filtration and the filtrate was concentrated in vacuo to afford the crude product as a brown solid. Trituration of the brown solid with dichloromethane afforded 292mg (50%) of the intermediate 4- (4-methoxyphenylamino) -7-chlorofuro [2, 3-d ] confirmed by LC/MS and NMR]Pyridazine. A suspension of this intermediate (292mg, 1.06mmol, 1 eq), (2-methylaminocarbonyl-4-pyridyl) methanol (intermediate H, 529mg, 3.18mmol, 3 eq) and 18-crown-6 (42mg, 0.16mmol, 15 mol%) in toluene (4mL) was stirred at room temperature for 20 min. KOH (178mg, 3.18mmol, 3 equiv.) was then added and the reaction mixture was heated at 80 ℃ for 36 h. After cooling to room temperature, water (10mL) was added and the mixture was stirred vigorously until a fine white suspension formed. Will be provided withThe suspension is filtered and washed with water and diethyl ether to yield 125mg (29%) of the desired product as a pale yellow solid: (M + H)⁺406；R_f0.50 (100% ethyl acetate).

Example 21: preparation of 4- (4-methoxyphenylamino) -7- (4-pyridylmethoxy) furo [2, 3-d ] pyridazine

The title compound was prepared using the procedure used for the preparation of example 20 substituting 4-pyridylmethanol for (2-methylaminocarbonyl-4-pyridyl) methanol. The pure product was isolated by flash column chromatography: (M + H)⁺349；R_f0.3 (95: 5 dichloromethane/methanol).

Example 22: preparation of 4- (4-methoxyphenylamino) -7- (2-aminocarbonyl-4-pyridylmethoxy) furo [2, 3-d ] pyridazine

The title compound was prepared using the procedure used for the preparation of example 16 substituting the product of example 21 for the product of example 9. The reaction was carried out with 250mg of starting material and proportional amounts of solvent and reagents. The crude product was purified by HPLC on a 75 x 30mmC18 column eluting with a linear gradient of 10-100% acetonitrile in water containing 0.1% trifluoroacetic acid at a rate of 10 ml/min for 10 minutes to afford 16mg of the title compound as a yellow solid: HPLC (50X 4.6mm YMCC18 column, linear gradient elution with a mixture of 10-100% acetonitrile in water containing 0.1% trifluoroacetic acid at a rate of 3 ml/min for 5 minutes, UV detection at 254 nm) for 1.98 minutes, peak；MS ES 392(M+H)⁺。

Examples 23 to 80: preparation of Compounds of the invention by Processes A-1, A-2 and A-3

Method A-1: the equivalent of dichloride (1) and M-NH are reacted₂Reflux in appropriate amount of absolute ethanol at 95 ℃ for 2 hours. The reaction mixture was cooled to room temperature, and the formed precipitate (2) was filtered and washed with isopropanol, 4.0N KOH, water and hexane in this order, and then dried. The filtrate (2) is then admixed with 1.2 equivalents of Q-NH₂The reaction was carried out in an appropriate amount of n-butanol at 150 ℃ for 10 hours. The reaction was cooled to room temperature and the solvent was evaporated under reduced pressure. The residue was treated with 4.0N aqueous KOH and extracted with dichloromethane. The organic layer was dried (magnesium sulfate) and evaporated. The crude product (3) was purified by preparative Thin Layer Chromatography (TLC) or flash silica gel chromatography eluting with methylene chloride/methanol (95: 5). The final product was confirmed by LC/MS and/or NMR. The following table shows that the compounds of the present invention of examples 23-25, 48, and 76-80 were prepared by the method A-1.

Method A-2: 1 equivalent of dichloride (1) is mixed with 2.2 equivalents of M-NH₂Reflux in an appropriate amount of n-butanol at 150 ℃ for 10 hours. The reaction mixture was cooled to room temperature, and the formed precipitate (4) was filtered and washed with isopropanol, 4.0N KOH, water and hexane in this order, and then dried. The crude product (4) was purified by preparative TLC or flash silica gel chromatography eluting with methylene chloride/methanol (95: 5). The final product was confirmed by LC/MS and/or NMR. The compounds of the invention of examples 26-33 and 75 shown in the following table were prepared by method A-2.

Method A-3: 1 equivalent of dichloride (1) and 1 equivalent of M-NH₂Suspended in DME (0.3M) and water was added until a solution was formed. The reaction mixture was heated at 65 ℃ for 48 hours. After cooling to room temperature, the precipitate formed was filtered off and washed with DME to obtain intermediate product (2) confirmed by LC/MS and NMR. In some cases, intermediate (2) was further purified by preparative TLC or washing with other solvents. A suspension of (2) (1 eq), methanol (3) (3 eq), and 18-crown-6 (10 mol%) in toluene (0.3M) was stirred at room temperature for 10 minutes. KOH (3 equivalents) was then added and the reaction mixture was heated at 80 ℃ for 24 hours. After cooling to room temperature, water was added and the mixture was stirred vigorously until a suspension formed. The suspension was filtered and washed with water to obtain the desired product (4). In some cases preparative TLC and/or washing with other solvents was used to further purify the final product. The final product was determined by LC/MS and NMR spectroscopy. The final product was confirmed by LC/MS and NMR. The following table shows that the compounds of the present invention of examples 34-47, 49-74, and 81-82D were prepared by method A-3.

Compounds prepared by parallel method A-1, A-2 or A-3

All compounds in the table were characterized by HPLC-positive ion electrospray mass spectrometry (HPLCES-MS, conditions below). In addition, some compounds were characterized by TLC on silica gel plates and showed R_fValue and solvent. For the other examples in the table, HPLC retention times are given;^aHPLC-electrospray mass spectrometry (HPLC ES-MS) was obtained using a Hewlett-Packard1100HPLC equipped with a quaternary pump, a variable wavelength detector, a YMC Pro C182.0 mm X23 mm column, and a Finnigan LCQ ion trap mass spectrometer with electrospray ionization. Elute on HPLC with a gradient of 90% a-95% B for 4 min. Buffer a was 98% water, 2% acetonitrile and 0.02% TFA. Buffer B was 98% acetonitrile, 2% water and 0.018% TFA. Scanning the spectra from 140-1200amu using different ion times depending on the number of ions in the ion source;^bin addition to HPLC ES-MS experiments, HPLC measurements for UV peak detection were also performed, and the barsThe parts are as follows: 50X 4.6mm YMCA C18 column eluted with a linear gradient of a mixture of 10-100% acetonitrile in water containing 0.1% trifluoroacetic acid at a rate of 3 ml/min for 5 minutes with UV detection at 254 nm;^cthe product was purified by RP-HPLC on a C18 column, with gradient elution using water/acetonitrile containing trifluoroacetic acid, which was added so that the trifluoroacetate salt could be isolated by evaporation of the pure product;^das noted, 4-pyridylcarbinol was used in step 2 of Process A-1 instead of the amine;^efor the preparation of 5-amino-2, 3-dihydrobenzofuran, see Mitchell, h.; leblanc, Y.J.org.chem.1994, 59, 682-687.^fReferences for the preparation of known TBS protected alcohol intermediates are: parsons, a.f.; pettifer, r.m.j.chem.soc.perkin trans.1, 1998, 651.

A moiety of the formula

The deprotection is carried out according to the following method: a solution of 3 equivalents of 1.0M TBAF in THF was added to a solution of the protected alcohol in THF (0.05M) at room temperature. The reaction mixture was stirred at room temperature for 1 hour, quenched with water, and then extracted with ethyl acetate.

Examples 83 to 92: preparation of isoquinoline by Process B-1

Method B-1: the dibromo isoquinoline (5.29mg, 0.1mmol) of step 1 of example 1 and M-NH in an 8-mL vial₂(0.2mmol) was heated in 1mL of n-butanol at 90 ℃ for 36 hours. Mixing the above materialsThe mixture was cooled to room temperature and the solvent was evaporated under reduced pressure. To the vial were added 4-mercaptopyridine (23mg, 0.2mmol) and cesium carbonate (67mg, 0.2 mmol). The mixture was heated at 180 ℃ for 1 hour and cooled to room temperature. Methanol (2mL) was added to the vial and the mixture was sonicated for 10 minutes and filtered. The methanol solution of the reaction mixture was collected and evaporated under reduced pressure. The formation of the product was confirmed by LC/MS. The compounds according to the invention of examples 83 to 92 shown in the table below were prepared by method B-1.

The compound prepared by Process B-1:

HPLC-electrospray mass spectrometry (HPLC ES-MS) was obtained with a Hewlett-Packard1100HPLC equipped with a quaternary pump, a variable wavelength detector, a YMC Pro c182.0 mm x 23mm column, and a finnigan lcq ion trap mass spectrometer with electrospray ionization. Elute on HPLC with a gradient of 90% a-95% B for 4 min. Buffer a was 98% water, 2% acetonitrile and 0.02% TFA. Buffer B was 98% acetonitrile, 2% water and 0.018% TFA. Spectra were scanned from 140-1200amu using different ion times depending on the number of ions in the ion source.

Examples 93 to 105: preparation of the novel phthalazine Compounds of the invention by parallel Synthesis

The novel phthalimide compounds 93-105 of the present invention are prepared from 1, 4-dichlorophthalazine (for its preparation see Novartis patent WO 98/35958, 11.02.98) instead of dichloroheterocyclopyridazine and the appropriate bicyclic compound and substituted aniline using method A-1 or A-2 as described above.

Novel phthalazines prepared by method A-1 or A-2:

Example 106-114: preparation of the salt of example 14

The product from example 14 (1.50g, 3.66mmol) was stirred as a slurry in methanol (20ml) and a solution of toluenesulfonic acid hydrate (0.701g, 3.67mmol) in methanol (5ml plus 5ml wash) was added dropwise rapidly. All materials dissolved within 5 minutes to give a yellow solution. Anhydrous ether (30ml) was added and stirring was continued for 5 minutes until a solid began to precipitate. The resulting mixture was quenched by stirring in an ice/water bath for 45 minutes, then the solid title product was collected by filtration (example 104), washed with diethyl ether, dried in a vacuum oven at 55 ℃ until no solvent was present by NMR analysis (1.5 hours). Other compounds were prepared in a similar manner by using a different acid than toluene sulfonic acid. Scaling up and using less methanol in the first step generally resulted in faster salt precipitation, and the use of a different solvent than that indicated for diethyl ether may help to crystallize out the individual salts. In some cases, methanol is first removed by evaporation in vacuo. Finally drying for 1.5 hours to several days depending on the amount of substance and the specific acid used.

The salt of example 14 was prepared

For HCl, the di-salt was isolated instead of the 1: 1 salt. This is also true if less than 2 equivalents of acid are used.

Bioassay and in vitro test data

KDR determination:

the cytoplasmic kinase domain of KDR kinase was expressed as a 6His fusion protein in Sf9 insect cells. The KDR domain of the fusion protein was purified using a Ni + + chelating column. 96-well ELISA plates were coated overnight at 4 ℃ with 5. mu.g of poly (G1u 4; Tyrl) (Sigma Chemical Co., St Louis, Mo.) in 100. mu.l of HEPES buffer (20mM HEPES, pH7.5, 150mM NaCl, 0.02% thimerosal). Before use, the plates were washed with HEPES, NaCl buffer and blocked with a solution of 1% BSA, 0.1% tween 20 in HEPES, NaCl buffer.

Test compounds were diluted from 4mM to 0.12. mu.M in 100% DMSO at half-log dilutionSerial dilutions were made. These dilutions were further diluted 20-fold in water to obtain compound solutions in 5% DMSO. Mu.l assay buffer (20mM HEPES, pH7.5, 100mM KCl, 10mM MgCl) was used₂，3mM MnCl₂0.05% glycerol, 0.005% Triton X-100, 1mM mercaptoethanol, with or without 3.3. mu.M ATP) loading assay plate 5. mu.l of diluted compound was added to 100. mu.l of the final assay solution. The final concentration was 10. mu.M-0.3 nM (in 0.25% DMSO). The assay was started by adding 10. mu.l (30ng) of KDR kinase domain.

The assay was incubated with test compound or vehicle under gentle agitation for 60 minutes at room temperature, the wells were washed with Phosphotyrosine (PY), and probed with anti-Phosphotyrosine (PY), mAb clone 4G10(Upstate Biotechnology, Lake plate, NY). The PY/anti-PY complex was detected with an anti-mouse IgG/HRP conjugate (Amersham International plc, Buckinghamshire, England). Phosphotyrosine was quantitated by incubation with 100. mu.l of 3, 3 ', 5, 5' -tetramethylbenzidine solution (Kirkegaard and Perry, TMB Microwell 1 Component peroxidase substrate). The color development was stopped by adding 100. mu.l of a 1% HCl based stop solution (Kirkegaard and Perry, TMB 1 Component StopSolution).

The optical density was determined spectrophotometrically at 450nm in a 96-well plate reader SpetraMax 250(Molecular Devices). Background (no ATP in the assay) OD values were subtracted from all ODs and percent inhibition was calculated according to the following formula:

% inhibition ═ [ (OD (vehicle control) -OD (with compound)) × 100]/[ OD (vehicle control) -OD (no ATP added) ]

Determination of IC Using Compound concentration versus percent inhibition Using least squares analysis₅₀The value is obtained. In this assay IC₅₀Compounds of ≦ 100nM include examples 1, 2, 4, 6, 8, 9, 10, 11, 12, 13, 14, 16, 17, 18, 19, 20, 22, 23, 24, 34, 37, 38, 39, 40, 42, 43, 44, 47, 49, 51, 52, 53, 54, 56, 57, 59, 60, 62, 63, 65, 66, 68, 69, 70, 71, 72, 73, 74, 7578, 82B, 82C, 82D, 85, 88, 93, 96, 97, 98, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, and 112. IC (integrated circuit)₅₀Compounds with values of 100nM-1000nM include the compounds of examples 3, 5, 7, 21, 27, 28, 35, 36, 45, 46, 48, 50, 55, 58, 61, 64, 67, 76, 79, 82A, 89, 95, 99, and 100. IC (integrated circuit)₅₀Compounds with values > 1000nM include the compounds of examples 26, 29, 30, 31, 32, 33, 41, 77, 80, 81, and 94. The compounds of the examples not listed can be assumed to have weak activity, their IC₅₀The value is greater than 1. mu.M.

Cell mechanistic assay-inhibition of 3T3KDR phosphorylation

NIH3T3 cells expressing the full-length KDR receptor were grown in DMEM (Life Technologies, inc., Grand Island, NY) supplemented with 10% neonatal calf serum, low glucose, 25mM/L sodium pyruvate, pyridoxine hydrochloride and 0.2mg/ml G418(Life Technologies, inc., Grand Island, NY). Cells were placed in collagen I-coated T75 flasks (Becton Dickinson Labware, Bedford, Mass.) in humidified 5% CO₂Maintained at 37 ℃ under an atmosphere.

15000 cells were plated into DMEM growth medium in each well of collagen I-coated 96-well plates. After 6 hours, the cells were washed and the medium was replaced with DMEM without serum. After overnight culture to quiesce the cells, the medium was replaced with Dulbecco's phosphate buffered saline (life technologies, inc., Grand Island, NY) containing 0.1% bovine albumin (sigma chemical co., St Louis, MO). After adding different concentrations (0-300nM) of test compound to the cells (1% final concentration in DMSO), the cells were incubated at room temperature for 30 min. The cells were then treated with VEGF (30ng/ml) for 10 min at room temperature. After VEGF stimulation, the buffer was removed and the cells were washed free by adding 150. mu.l of extraction buffer (50mM Tris, pH7.8, supplemented with 10% glycerol, 50mM BGP, 2mM EDTA, 10mM NaF, 0.5mM NaVO)₄And 0.3% TX-100) cells were lysed at 4 ℃ for 30 minutes.

To evaluate receptor phosphorylation, 100 microliters of each cell lysate was added to wells of ELISA plates pre-coated with 300ng of antibody C20(Santa Cruz Biotechnology, inc., Santa Cruz, CA). After incubation for 60 min, the plates were washed and the phosphotyrosine of the bound KDR was probed with anti-phosphotyrosine mAb clone 4G10(Upstate Biotechnology, Lake plain, NY). Plates were washed and each well was incubated with anti-mouse IgG/HRP conjugate (Amersham International plc, Buckinghamshire, England) for 60 minutes. The wells were washed and phosphotyrosine was quantitated by adding 100. mu.l of 3, 3 ', 5, 5' -tetramethylbenzidine solution (Kirkegaard and Perry, TMB Microwell 1 Component peroxidase substrate) to each well. The color development was stopped by adding 100. mu.l of a 1% HCl based stop solution (Kirkegaard and Perry, TMB 1 Component StopSolution).

Optical Density (OD) was determined spectrophotometrically at 450nm in a 96-well plate reader (SpetraMax 250, Molecular Devices). Background (no VEGF added) OD values were subtracted from all ODs and percent inhibition was calculated according to the following formula:

% inhibition ═ [ (OD (VEGF control) -OD (with compound)) × 100]/[ OD (VEGF control) -OD (without VEGF added) ]

Using compound concentration versus percent inhibition, the IC of certain example compounds was determined using least squares analysis₅₀In this assay IC₅₀Compounds of ≦ 20nM include the compounds of examples 2, 6, 10, 11, 14, 23, 96, 101, 102, 103, 104, 105. IC (integrated circuit)₅₀Compounds with values of 20nM to 50nM include the compounds of examples 1, 4, 8, 9, 12, 13, 17, 24, 93, 98. IC (integrated circuit)₅₀Compounds with values of 50nM to 400nM include the compounds of examples 97, 99, and 100.

An angiogenesis model:

preparation of Matrigel plugs and in vivo phase:(collagen biological Products, Bedford, MA) is a basement membrane extracted from murine tumors, which is composed mainly of laminin, collagen IV and heparan sulfate proteoglycans. It is provided as a sterile liquid at 4 ℃, but rapidly forms a solid gel at 37 ℃.

Liquid Matrigel was mixed with SK-MEL2 human tumor cells transfected with a plasmid containing the murine VEGF gene with a selectable marker at 4 ℃. Tumor cells were grown in vitro under selection and the cells were mixed with cold liquid Matrigel at 2X 10⁶Mixed at a ratio of 0.5 ml. 0.5ml was implanted subcutaneously near the midline of the abdomen with a 25 gauge needle. Starting on the day of implantation, test compounds were administered orally as solutions in ethanol/Cremaphor EL/saline (12.5%: 75%) once daily at doses of 30, 100, and 300 mg/kg. 12 days after implantation, the mice were euthanized and Matrigel pellets were collected for analysis of hemoglobin content.

And (3) hemoglobin measurement: the Matrigel pellets were placed in 4 volumes (w/v) of lysis buffer (20mM Tris pH7.5, 1mM EGTA, 1mM EDTA, 1% Triton X-100[ EM Science, Gibbstown, N.J. ], and a complete, EDTA-free protease inhibitor cocktail [ Mannheim, Germany ]) at 4 ℃ and homogenized at 4 ℃. The homogenate was incubated on ice for 30 minutes with shaking and centrifuged at 14 kXg for 30 minutes at 4 ℃. The supernatant was transferred to a chilled microfuge tube and stored at 4 ℃ for hemoglobin determination.

Mouse hemoglobin (Sigma Chemical co., st. louis, MO) was suspended at a concentration of 5mg/ml in autoclaved water (BioWhittaker, Inc, walker, MD.). Standard curves were generated from 500. mu.g/ml to 30. mu.g/ml in lysis buffer (see above). Standard curve and lysate samples were added in duplicate, 5 μ l/well to polystyrene 96-well plates. TMB substrate was resuspended in 50ml of acetic acid solution at room temperature using Sigma Plasma hemoglobin kit (Sigma Chemical co., st. To each well 100 microliters of substrate was added followed by 100 microliters/well hydrogen peroxide solution at room temperature. The plates were incubated at room temperature for 10 minutes.

Optical density was determined spectrophotometrically at 600nm in a 96-well plate reader SpetraMax 250 microplate spectrophotometer System (Molecular Devices, Sunnyvale, Calif.). Background lysis buffer readings were subtracted from all wells.

The total sample hemoglobin content was calculated according to the following formula:

total hemoglobin (sample lysate volume) x (hemoglobin concentration)

The average total hemoglobin of the Matrigel samples without cells was subtracted from the total hemoglobin of each cell-containing Matrigel sample. Percent inhibition was calculated according to the following formula:

% inhibition (mean total hemoglobin drug-treated tumor lysate) x 100/(mean total hemoglobin untreated tumor lysate)

The compound of example 8 administered orally at doses of 100 and 300mg/kg showed significant activity in this assay, with total hemoglobin content of Matrigel samples from the animals administered being suppressed by > 60% relative to vehicle control animals. No other example compounds were tested in this model.

Other embodiments of the invention will be apparent to those skilled in the art from consideration of the specification or practice of the invention disclosed herein. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the invention being indicated by the following claims.

Claims

1.A compound having the general formula or a pharmaceutically acceptable salt thereof

Wherein

R¹And R²Together form a bridging group of the structure

Wherein the attachment is via a terminal carbon atom;

and wherein

m is 0 or 1;

G¹is a substituent independently selected from the group consisting of:

a halogen;

·(C₁-C₅) An alkyl group;

halogen substituted (C)₁-C₅) An alkyl group;

hydroxy-substituted (C)₁-C₅) An alkyl group;

·-OR⁶；

halo (C)₁-C₅) An alkoxy group;

·-OCOR⁶；

·-COR⁶；

·-CO₂R⁶；

·-CON(R⁶)₂；

·-CH₂OR³(ii) a And

·-OCON(R⁶)₂；

R³is H or (C)₁-C₅) An alkyl group;

R⁶is independently selected from

H; and

·(C₁-C₅) An alkyl group;

R⁴is H;

x is NH;

y is (C)₁-C₅) An alkylene group;

z is CR⁴；

p is 0;

q is 1 or 2;

G³is an amide of the formula

·-CON(R⁶)₂；

A and D independently represent N or CH;

b and E independently represent N or CH;

l represents N or CH;

with the following conditions:

a) in the ring containing A, B, D, E, and L, the total number of N atoms is 1; and

b) when L represents CH, and any G³When it is a monovalent substituent, at least one of A and D is an N atom;

j is a phenyl ring;

q' represents a substituent G on the ring J⁴And is 0, 1, 2 or 3; and is

G⁴Is a monovalent or divalent group selected from:

·-N(R⁶)₂；

a halogen;

·(C₁-C₅) An alkyl group;

halogen substituted (C)₁-C₅) An alkyl group;

·-OR⁶(ii) a And

a bivalent bridging group forming a fused ring attached to and connecting adjacent rings J, said bridging group having the following structure:

a)

wherein

Each T²Independently represents N or CH;

T³represents S or O; wherein

Via the terminal atom T²And T³To ring J;

b)

wherein each T⁵And T⁶Independently represent O, S, C (R)⁴)₂Or CF₂(ii) a Wherein

Via the terminal atom T⁵To ring J;

with the following conditions:

i) comprising T⁵And T⁶The bridging group of atoms may contain up to two heteroatoms O or S; and

ii) in a solvent system comprising T⁵And T⁶In the bridging group of the atom, when a T⁵And a T⁶Is an O atom, or two T⁶When is an O atom, said O atoms are separated by at least one carbon atom;

further conditions are:

at G¹、G²、G³And G⁴In (2), when two radicals R are present³Or R⁶Are each alkyl and, when located on the same N atom, may be bonded via a bond, O, S, or NR³Linked to form a nitrogen-containing heterocycle having 5-7 ring atoms.

2. A pharmaceutical composition comprising a compound of claim 1 and a pharmaceutically acceptable carrier.

3. Use of a compound of claim 1 in the manufacture of a medicament for treating a mammal suffering from a condition characterized by abnormal angiogenesis or hyper-osmotic processes.

4. The use of claim 3, wherein the disorder is tumor growth; retinopathy; rheumatoid arthritis; psoriasis; or bullous diseases associated with epidermal blister formation.

5. The use of claim 4, wherein the retinopathy is selected from the group consisting of diabetic retinopathy, ischemic retinal-vein occlusion, retinopathy of prematurity, and age-related macular degeneration.

6. The use of claim 4, wherein the bullous disease is selected from the group consisting of bullous pemphigoid, erythema multiforme, and dermatitis herpetiformis.

7. A compound selected from the group consisting of:

4- ({1- [ (4-chlorophenyl) amino ] -4-isoquinolinyl } methyl) -2-pyridinecarboxamide; and

4- ({1- [ (4-chlorophenyl) amino ] -4-isoquinolinyl } methyl) -N-methyl-2-pyridinecarboxamide.