AU2012204137A1 - Organic compounds - Google Patents

Abstract

1- or 2-substituted (6aR,9aS)-3-(phenylamino)-5-6a,7,8,9,9a-hexahydro-5-methyl cyclopent[4,5]imidazo[ 1,2-a]pyrazolo[4,3-e]pyrimidin-4(JH or 2H)-one compounds of Formula (I), processes for their production, their use as pharmaceuticals and pharmaceutical compositions comprising them.

Description

AUSTRALIA Patents Act COMPLETE SPECIFICATION (ORIGINAL) Class Int. Class Application Number: Lodged: Complete Specification Lodged: Accepted: Published: Priority Related Art: Name of Applicant: Takeda Pharmaceutical Company Limited Actual Inventor(s): Peng Li Address for Service and Correspondence: PHILLIPS ORMONDE FITZPATRICK Patent and Trade Mark Attorneys 367 Collins Street Melbourne 3000 AUSTRALIA Invention Title: ORGANIC COMPOUNDS Our Ref: 945825 POF Code: 474551/483471 The following statement is a full description of this invention, including the best method of performing it known to applicant(s): -1 - ORGANIC COMPOUNDS The present application is a divisional application of Australian patent application number 2008335811, which claims priority from U.S. Provisional application No. 61/012,040, filed December 6, 2007, the contents of which are hereby incorporated by reference. 5 TECHNICAL FIELD [00011 The present invention relates to 1- or 2-substituted (6aR*,9aS*)-3-(phenylamino) 5-6a,7,8,9,9a-hexahydro-5-methyl-cyclopent[4,5]imidazo[1,2-a]pyrazolo[4,3-e]pyrimidin-4(1H or 2H)-one compounds, preferably I- or 2-substituted (6aR,9aS) (phenylamino)-5-6a,7,8,9,9a 10 hexahydro-5-methyl-cyclopent[4,5]imidazo[ I,2-a]pyrazolo[4,3-e]pyrimidin-4(JH or 2H)-one compounds of Formula I as described below, processes for their production, their use as pharmaceuticals and pharmaceutical compositions comprising them. Of particular interest are novel compounds useful as inhibitors of phosphodiesterase I (PDEI), e.g., in the treatment of diseases involving disorders of the dopamine Dl receptor intracellular pathway, such as 15 Parkinson's disease, depression, narcolepsy, damage to cognitive function, e.g., in schizophrenia, or disorders that may be ameliorated through enhanced progesterone-signaling pathway, e.g., female sexual dysfunction. BACKGROUND OF THE INVENTION 20 [00021 Eleven families of phosphodiesterases (PDEs) have been identified but only PDEs in Family I, the Ca"-calmodulin-dependent phosphodiesterases (CaM-PDEs), have been shown to mediate both the calcium and cyclic nucleotide (e.g. cAMP and cGMP) signaling pathways. The three CaM-PDE genes, PDE I A, PDE1B, and PDEI C, are all expressed in central nervous system tissue. PDEI A is expressed throughout the brain with higher levels of expression in the CAI to 25 CA3 layers of the hippocampus and cerebellum and at a low level in the striatum. PDEIA is also expressed in the lung and heart. PDE 11B is predominately expressed in the striatum, dentate gyrus, olfactory tract and cerebellum, and its expression correlates with brain regions having high levels of dopaminergic innervation. Although PDEI B is primarily expressed in the central nervous system, it may be detected in the heart. PDE1C is primarily expressed in olfactory epithelium, 30 cerebellar granule cells, and striatum. PDEIC is also expressed in the heart and vascular smooth muscle. [0003] Cyclic nucleotide phosphodiesterases decrease intracellular cAMP and cGMP signaling by hydrolyzing these cyclic nucleotides to their respective inactive 5'-monophosphates (5'AMP and 5'GMP). CaM-PDEs play a critical role in mediating signal transduction in brain 35 cells, particularly within an area of the brain known as the basal ganglia la or striatum. For example, NMDA-type glutamate receptor activation and/or dopamine D2 receptor activation result in increased intracellular calcium concentrations, leading to activation of effectors such as calmodulin-dependent kinase II (CaMKII) and calcineurin and to activation of CaM-PDEs, resulting in reduced cAMP and cGMP. Dopamine DI receptor 5 activation, on the other hand, leads to activation of nucleotide cyclases, resulting in increased cAMP and cGMP. These cyclic nucleotides in turn activate protein kinase A (PKA; cAMP dependent protein kinase) and/or protein kinase Q (PKG; cGMP-dependent protein kinase) that phosphorylate downstream signal transduction pathway elements such as DARPP-32 (dopamine and cAMP-regulated phosphoprotein) and cAMP responsive element binding 10 protein (CREB). Phosphorylated DARPP-32 in turn inhibits the activity of protein phosphates-I (PP-1), thereby increasing the state of phosphorylation of substrate proteins such as progesterone receptor (PR), leading to induction of physiologic responses. Studies in rodents have suggested that inducing cAMP and cGMP synthesis through activation of dopamine Dl or progesterone receptor enhances progesterone signaling associated with 15 various physiological responses, including the lordosis response associated with receptivity to mating in some rodents. See Mani, et al., Science (2000) 287: 1053. 100041 CaM-PDEs can therefore affect dopamine-regulated and other intracellular signaling pathways in the basal ganglia (striatum), including but not limited to nitric oxide, noradrenergic, neurotensin, CCK, VIP, serotonin, glutamate (e.g., NMDA receptor, AMPA 20 receptor), GABA, acetylcholine, adenosine (e.g., A2A receptor), cannabinoid receptor, natriuretic peptide (e.g., ANP, BNP, CNP), DARPP-32, and endorphin intracellular signaling pathways. 100051 Phosphodiesterase (PDE) activity, in particular, phosphodiesterase 1 (PDEI) activity, functions in brain tissue as a regulator of locomotor activity and learning and 25 memory. PDEI is a therapeutic target for regulation of intracellular signaling pathways, preferably in the nervous system, including but not limited to a dopamine Dl receptor, dopamine D2 receptor, nitric oxide, noradrenergic, neurotensin, CCK, VIP, serotonin, glutamate (e.g., NMDA receptor, AMPA receptor), GABA, acetylcholine, adenosine (e.g., A2A receptor), cannabinoid receptor, natriuretic peptide (e.g., ANP, BNP, CNP) , endorphin 30 intracellular signaling pathway and progesterone signaling pathway. For example, inhibition of PDEIB should act to potentiate the effect of a dopamine DI agonist by protecting cGMP and cAMP from degradation, and should similarly inhibit dopamine D2 receptor signaling pathways, by inhibiting PDEI activity. Chronic elevation in intracellular calcium levels is linked to cell death in numerous disorders, particularly in neurodegerative diseases such as 2.

Alzheimer's, Parkinson's and Huntington's Diseases and in disorders of the circulatory system leading to stroke and myocardial infarction. PDE1 inhibitors are therefore potentially useful in diseases characterized by reduced doparnine Dl receptor signaling activity, such as Parkinson's disease, restless leg syndrome, depression, narcolepsy and cognitive impairment. PDE1 5 inhibitors are also useful in diseases that may be alleviated by the enhancement of progesterone-signaling such as female sexual dysfunction. There is thus a need for compounds that selectively inhibit PDE1 activity, especially PDElB activity. SUMMARY OF THE INVENTION 10 In one preferred embodiment, the present invention provides a compound of formula Q

RA

4 0 N.-R 5 N N N/X-Y Formula Q wherein (i) -X-Y-Z is selected from 4-(5-fluoropyrid-2-yl)benzyl, 4-(6-fluoropyrid-2 15 yl)benzyl, 4-(3-fluoropyrid-2-yl)benzyl, 4-(4-fluoropyrid-2-yl)benzyl; (ii) R4 and R5 are independently H, C 1 -alky or aryl optionally substitute with one or more halo, hydroxy or Ci..alkoxy; in free or salt form. In another preferred embodiment, the present invention provides 1- or 2-substituted 20 (6aR*,9aS*)-3-(phenylamino)-5-6a,7,8,9,9a-hexahydro-5-methyl-cyclopent[4,5]imidazo[ 1,2 a]pyrazolo[4,3-e]pyrimidin-4(lH or 2H)-one compounds, preferably 1- or 2-substituted (6aR,9aS)-3-(phenylamino)-5-6a,7,8,9,9a-hexahydro-5-methyl-cyclopent[4,5]imidazo[ 1,2 a]pyrazolo[4,3-e]pyrimidin-4(lH or 2H)-ones, of formula Q 3 R 4 O N R 5 NI N! N N1 N X-Y Formula Q wherein (i) X is Ci-6alkylene (e.g., methylene, ethylene or prop-2-yn-I-ylene); 5 (ii) Y is a single bond, alkynylene (e.g., -CEC-), arylene (e.g., phenylene) or heteroarylene (e.g., pyridylene); (iii) Z is H, aryl (e.g., phenyl), heteroaryl (e.g., pyridyl, e.g., pyrid-2-yl), halo (e.g., F, Br, Cl), haloCI.

6 alkyl (e.g., trifluoromethyl), -C(O)-R', N(R 2

)(R

3 ), or C3.7cycloalkyl optionally containing at least one atom 10 selected from a group consisting of N or 0 (e.g., cyclopentyl, cyclohexyl, tetrahydro-2H-pyran-4-yl, or morpholinyl); (iv) R' is CI.

6 alkyl, haloC 3 -6alkyl, -OH or -OCi-6alkyl (e.g., -OCH 3 ); (v) R 2 and R 3 are independently H or Ci.

6 alkyl; (vi) R 4 and R 5 are independently H, Ci-6alky or aryl (e.g., phenyl) optionally 15 substituted with one or more halo (e.g., fluorophenyl, e.g., 4 fluorophenyl), hydroxy (e.g., hydroxyphenyl, e.g., 4-hydroxyphenyl or 2 hydroxyphenyl) or Ci.

6 alkoxy; (vii) wherein X, Y and Z are independently and optionally substituted with one or more halo (e.g., F, Cl or Br), CI.

6 alkyl (e.g., methyl), haloCI.

6 alkyl 20 (e.g., trifluoromethyl), for example, Z is heteroaryl, e.g., pyridyl substituted with one or more halo (e.g., 6-fluoropyrid-2-yl, 5-fluoropyrid 2-yl, 6-fluoropyrid-2-yi, 3-fluoropyrid-2-yl, 4-fluoropyrid-2-yl, 4,6 dichloropyrid-2-yl), haloC.

6 alkyl (e.g., 5-trifluoromethylpyrid-2-yl) or CI. 4 6 alkyl (e.g., 5-methylpyrid-2-yl), or Z is aryl, e.g., phenyl, substituted with one or more halo (e.g., 4-fluorophenyl), in free, salt or prodrug form. In a further embodiment, the invention provides a compound of Formula Q, which includes the proviso that when X is an unsubstituted 5 methylene, Y is phenylene or heteroarylene, and Z is aryl, heteroaryl, haloalkyl or cycloalkyl, then Z is substituted with at least one halo (e.g., fluoro, chloro, bromo) or alkyl (e.g., methyl, ethyl) group. [00081 The invention further provides compounds of Formula Q as described above as follows: 10 1.1 Formula Q, wherein X is Ci.6alkylene (e.g., methylene, ethylene or prop 2-yn-I-ylene) optionally substituted with one or more halo (e.g., F, Cl or Br), Ci.

6 alkyl (e.g., methyl), haloC,.

6 alkyl (e.g., trifluoromethyl); 1.2 Formula Q or 1.1, wherein X is methylene or ethylene optionally substituted with one or more halo (e.g., F, Cl or Br), CI.

6 alkyl (e.g., 15 methyl), haloC.

6 alkyl (e.g., trifluoromethyl); 1.3 Formula Q, 1.1 or 1.2, wherein X is methylene optionally substituted with one or more halo (e.g., F, Cl or Br), C 1

.

6 alkyl (e.g., methyl), haloCI- 6 alkyl (e.g., trifluoromethyl); 1.4 Formula Q, 1.1, 1.2 or 1.3, wherein X is methylene substituted with 20 bromo; 1.5 Formula Q, 1.1, 1.2, 1.3 or 1.4, wherein X is ethylene; 1.6 Formula Q or any of 1.1-1.5, wherein X is prop- 2 -yn-I-ylene; 1.7 Formula Q or any of 1.1-1.6, wherein Y is a single bond, arylene (e.g., phenylene) or heteroarylene (e.g., pyridylene) optionally substituted with 25 one or more halo (e.g., F, Cl or Br), CI.

6 alkyl (e.g., methyl), haloC 1 -6alkyl (e.g., trifluoromethyl); 1.8 Formula Q or any of 1.1-1.7, wherein Y is arylene (e.g., phenylene) optionally substituted with one or more halo (e.g., F, Cl or Br), C 1

.

6 alkyl (e.g., methyl), haloC 1

.

6 alkyl (e.g., trifluoromethyl); 30 1.9 Formula Q or any of 1.1-1.8 wherein Y is phenylene optionally substituted with fluoro at the 3 or 5-position of the phenylene ring; 5 1.10 Formula Q or any of 1.1-1.9, wherein Y is phenylene; 1.11 Formula Q or any of 1.1-1.7, wherein Y is heteroarylene (e.g., pyridylene) optionally substituted with one or more halo (e.g., F, Cl or Br), C1.

6 alkyl (e.g., methyl), haloCI.

6 alkyl (e.g., trifluoromethyl); 5 1.12 Formula Q or any of 1.1-1.7, wherein Y is a single bond; 1.13 Formula Q or any of 1.1-1.7, wherein Y is heteroaryl (e.g., pyrid-2-yl); 1.14 Formula Q or any of 1.1-1.13, wherein Z is H, aryl (e.g., phenyl), heteroaryl (e.g., pyridyl, e.g., pyrid-2-yl), halo (e.g., F, Br, Cl), haloCj. 6 alkyl (e.g., trifluoromethyl), -C(O)-R', -N(R 2

)(R

3 ), or C3.

7 cycloalkyl 10 optionally containing at least one atom selected from a group consisting of N or 0 (e.g., cyclopentyl, cyclohexyl, tetrahydro-2H-pyran-4-yl, or morpholinyl), wherein said aryl and heteroaryl are optionally substituted with one or more halo (e.g., F, Cl or Br), C1.

6 alkyl (e.g., methyl), haloCj. 6alkyl (e.g., trifluoromethyl); 15 1.15 Formula Q or any of 1.1-1.14, wherein Z is C 3 .7cycloalkyl optionally containing at least one atom selected from a group consisting of N or 0 (e.g., cyclopentyl, cyclohexyl, tetrahydro-2H-pyran-4-yl, morpholinyl); 1.16 Formula Q or any of 1.1-1.15, wherein Z is cyclopentyl; 1.17 Formula Q or any of 1.1-1.15, wherein Z is tetrahydro-2H-pyran-4-yl; 20 1.18 Formula Q or any of 1.1-1.14, wherein Z is heteroaryl (e.g., pyridyl, e.g., pyrid-2-yl) optionally substituted with one or more halo (e.g., F, Cl or Br), Ci-6alkyl (e.g., methyl), haloCI.6alkyl (e.g., trifluoromethyl); 1.19 Formula Q or any of 1.1 -1.14 or 1.18, wherein Z is pyridyl; 1.20 Formula Q or any of 1.1-1.14 or 1.18, wherein Z is pyrid-2-yl; 25 1.21 Formula Q or any of 1.1-1.14 or 1.18, wherein Z is 3-fluoropyrid-2-yl; 1.22 Formula Q or any of 1.1-1.14 or 1.18, wherein Z is 4-fluoropyrid-2-yl; 1.23 Formula Q or any of 1.1-1.14 or 1.18, wherein Z is 5-fluoropyrid-2-yl; 1.24 Formula Q or any of 1.1-1.14 or 1.18, wherein Z is 6-fluoropyrid-2-yl; 1.25 Formula Q or any of 1.1-1.14 or 1.18, wherein Z is heteroaryl, e.g., 30 pyridyl, optionally substituted with one or more haloCI6alkyl (e.g., 5 trifluoromethylpyrid-2-yl; 6 1.26 Formula Q or any of l.1-l.14 or 1.18, wherein Z is 5 trifl uoromethylpyrid-2-yl; 1.27 Formula Q or any of 1.1-1.14 or 1.18, wherein Z is heteroaryl, e.g., pyridyl, optionally substituted with one or more C 1

.

6 alkyl (e.g., 5 5 methylpyrid-2-yl); 1.28 Formula Q or any of 1.1-1.14 or 1.18, wherein Z is 5-methylpyrid-2-yl; 1.29 Formula Q or any of 1.1-1.14, wherein Z is haloC 1

.

6 alkyl (e.g., trifluoromethyl); 1.30 Formula Q or any of 1.1-1.14 or 1.29, wherein Z is trifluoromethyl; 10 1.31 Formula Q or any of 1.1-1.14 wherein Z is aryl, e.g., phenyl, optionally substituted with one or more halo (e.g., 4-fluorophenyl), haloC 1

.

6 alkyl or

C..

6 alkyl; 1.32 Formula Q or any of 1.1-1.14, wherein Z is aryl (e.g., phenyl) one or more halo (e.g., 4-fluorophenyl), haloC.

6 alkyl or CI-6alkyl; 15 1.33 Formula Q or any of 1.1-1.14 or 1.36, wherein Z is phenyl; 1.34 Formula Q or any of 1.1-1.14 or 1.31 wherein Z is 4-fluorophenyl; 1.35 Formula Q or any of 1.1-1.14, wherein Z is -C(O)-R' and R' is Ci.

6 alky (e.g., methyl), haloC.

6 alkyl (e.g., trifluoromethyl), -OH or -OCI.

6 alkyl (e.g., -OCH 3 ); 20 1.36 Formula Q or any of 1.1-1.14 or 1.29, wherein Z is-C(O)-R' and R' is methyl; 1.37 Formula Q or any of 1.1-1.14 or 1.29, wherein Z is-C(O)-R' and R' is trifluoromethyl; 1.38 Formula Q or any of 1.1-1.14 or 1.29, wherein Z is-C(O)-R' and R' is 25 OH; 1.39 Formula Q or any of 1.1-1.14 or 1.29, wherein Z is-C(O)-R' and R' is OCI.

6 alkyl (e.g., -OCH3); 1.40 Formula Q or any of 1.1-1.14 or 1.29, wherein Z is-C(O)-R' and R' is OCH 3 ; 30 1.41 Formula Q or any of 1.1-1.14, wherein Z is -N(R 2

)(R

3 ); 7 1.42 Formula Q or any of 1.l-1.14 or 1.32, wherein Z is -N(R 2

)(R

3 ), wherein R 2 and R 3 are methyl; 1.43 Formula Q or any of 1.1-1.42 wherein R 4 and R 5 are independently H, C1. 6 alky or aryl (e.g., phenyl) optionally substituted with one or more halo 5 (e.g., fluorophenyl, e.g., 4-fluorophenyl), hydroxy (e.g., hydroxyphenyl, e.g., 4-hydroxyphenyl or 2-hydroxyphenyl) or C 1

.

6 alkoxy; 1.44 Formula Q or any of 1.1-1.42 wherein either R 4 or R 5 is H; 1.45 Formula Q or any of 1.1-1.42 wherein either R 4 or R 5 is aryl (e.g., phenyl) optionally substituted with one or more halo (e.g., fluorophenyl, e.g., 4 10 fluorophenyl), hydroxy (e.g., hydroxyphenyl, e.g., 4-hydroxyphenyl or 2 hydroxyphenyl) or Cg.

6 alkoxy; 1.46 Formula Q or any of 1.1-1.42 wherein either R 4 or R 5 is aryl (e.g., phenyl) optionally substituted with one or more halo (e.g., fluorophenyl, e.g., 4 fluorophenyl), hydroxy (e.g., hydroxyphenyl, e.g., 4-hydroxyphenyl or 2 15 hydroxyphenyl) or CI.

6 alkoxy; 1.47 Formula Q or any of 1.1-1.42 wherein either R 4 or R 5 is phenyl; 1.48 Formula Q or any of 1.1-1.42 wherein either R 4 or R 5 is phenyl substituted with fluoro (e.g., 4-fluorophenyl); 1.49 Formula Q or any of 1.]-l .42 wherein either R 4 or R 5 is phenyl substituted 20 with hydroxy (e.g., 4-hydroxyphenyl) or CI.

6 alkoxy; 1.50 any of the preceding formulae wherein -X-Y-Z is selected from 4-(5 fluoropyrid-2-yl)benzyl, 4 -(6-fluoropyrid-2-yl)benzyl, 4-(3-fluoropyrid-2 yl)benzyl, 4

-(

6 -trifluoromethylpyrid-2-yl)benzyl, 4-(4-fluoropyrid-2 yl)benzyl, 4 -(S-methylpyrid-2-yl)benzyl, 4-(4-fluoropphenyl)benzyl, 25 biphenyl-4-ylmethyl, 4-trifluoromethylphenyl, 4-(4,6-dichloropyrid-2 yl)benzyl, 4-(pyrid-2-yl)benzyl, and 4-(carboxy)benzyl, 4 (methyicarboxy)benzyl; 1.51 any of the preceding formulae wherein R 4 is H and R 5 is phenyl; 1.52 Formula Q or any of 1.1-1.51, wherein R 4 is H and R 5 is 4-fluorophenyl or 30 4-hydroxyphenyl; 8 1.53 any of the preceding formulae wherein the compound is selected from a group consisting of: O HN \Q N )N Br 0 HN N N NN N N -- N N NN F 0 O HN OHN--I N N NNN N N N N N' Nv F F 9N F0 FF 0OHN\' 0OHN\' NN H N N FF N N N F 0 N N -0 HN-9 O HN-' O HN0 NN F . O N j N O N N F F "~F F 0OHN\00 N NNN N1 NNN 0OHN\I N N 0OHN\' NN N N N4N F F F 10 0N HN / N)' N 0 HN N\ N /\ N> N ~ii N/~ N 0 HN \/ N N 0 N \ N"" NN F F F, HN \ / 0 HN \ N N N NF F K F 0HN0 0 N N N N~~N >NF N~k N' N N' F 0 HN-j NN 0 HN-0 N N N/ W" N NF F, N N ]~N KNNN rN/ NN F $NF 0 I N N H-a N~"NN NN H N N;; N N N N N" N~ 0 "' NN ' N/MN 12 HO HNkI/j 0 N N N N N N N NN N N N 5 F N N N N N N N N 'N1 N 3 NN H ON N NNN N/\ F

HN\/

0 HNO/ N -Y HN~ \ /"N N N ' N N- F N F F F - 0 HN\ 0 HN \ /N Ni7 NN N, N , 0 - Q N( NN \ N SIN F N N N N~ N N bI~ N'~ 5 14 0

H

N N J N 0 HN- N" N N/ N 0~ N N N 0 N 47) N N N NN N N:N N / ooo '-N(/ FI N F F F 0 HN-0 ~ OH" NN HN< WN N N /c 15 HO 0 HN I HL ~N

HN

NN N/ N N N N O HN 0 HN o HNo HN NN N HN N6 N N 0 N 7 N 11 OH and' 1 .55 any of the preceding formulae wherein the compound is selected from a 5 group consisting of: NN N / NP F 0 HN\/ NN N Nb N: , N% N F 16 0HN \ "N 0 N \ / N< N N N 'N N -0 '~ F F o H\/ N N N 0 H N\ b~ N> F ~II~F F' o H NJ N ~ 0 HN\ N N N "N Fii N NN NF NN N 0 \ 0 HNC9 /' N\ N x NN F P N 7_ o HN0? 0NN 0 HN\ / N'N' N NIN' b N 0 HN4C9 H_ - F N N N- / N N N NN N F N' P F CI F NI - N N" N FNW'N N / 'zK) N 18 HO 0 HN NLN N N F s N HN N N N N N N OH , and ; 1.56 any of the preceding formulae wherein the compounds inhibit phosphodiesterase-mediated (e.g., PDE 1-mediated, especially PDE I B 5 mediated) hydrolysis of cGMP, e.g., with an IC 50 of less than I PM, preferably less than 200 nM, more preferably less than 100 nM, more preferably less than 50 nM, even more preferably less than 25 nM in an immobilized-metal affinity particle reagent PDE assay, for example, as described in Example 15, 10 in free, salt or prodrug form. In a further embodiment, the invention provides a compound of any of formulae 1.1-1.56, provided that when X is an unsubstituted methylene, Y is phenylene or heteroarylene, and Z is aryl, heteroaryl, haloalkyl or cycloalkyl, then Z is a substituted with at least one halo (e.g., fluoro, chloro, bromo) or alkyl (e.g., methyl, ethyl) group. 15 100091 The invention also provides formula 1 19 N HN N N X-Y z Formula I wherein (i) X is C 1

.

4 alkylene (e.g., methylene, ethylene or prop-2-yn-I-ylene); 5 (ii) Y is a single bond, alkynylene (e.g., -C=C-), arylene (e.g., phenylene) or heteroarylene (e.g., pyridylene); (iii) Z is H, aryl (e.g., phenyl), heteroaryl (e.g., pyrid-2-yl), halo (e.g., F, Br, Cl), haloC 1 4alkyl (e.g., trifluoromethyl), -C(O)-R', -N(R 2

)(R

3 ), or C 3 . 7 cycloalkyl optionally containing at least one atom selected from a group 10 consisting of N or 0 (e.g., cyclopentyl, cyclohexyl, tetrahydro-2H-pyran 4-yI, or morpholinyl); (iv) R' is C 1 4alkyl, haloC.4alkyl; (v) R 2 and R 3 are independently H or Ci.4alkyl, (vi) wherein X, Y and Z are independently and optionally substituted with halo 15 (e.g., F, Cl or Br), for example, Z is pyrid-2-yl substituted with fluoro (e.g., 6-fluoro-pyrid-2-yl), in free, salt or prodrug form, including its enantiomers, diastereoisomers and racemates, provided that when X is an unsubstituted methylene, Y is phenylene or heteroarylene, and Z is aryl, heteroaryl, haloalkyl or cycloalkyl, then Z is a substituted with at least one 20 halo (e.g., fluoro, chloro, bromo) or alkyl (e.g., methyl, ethyl) group. [00101 Preferably, (6aR,9aS)-3-(phenylamino)-5-6a,7,8,9,9a-hexahydro-5 methyl-cyclopent[4,5]imidazo[I,2-a]pyrazolo[4,3-e]pyrimidin-4(2H)-ones is 2 20 substituted, e.g., -X-Y-Z is substituted on the 2-position of the pyrazolo ring of Formula I, for example: OHN N-X N N N y-Z Formula II 5 100111 More preferably, when Y is phenylene and Z is substituted at the para position of the phenyl ring. [0012] The invention further provides compounds of Formula I as follows: 1.57 Formula I, wherein X is C 1 4 alkylene (e.g., methylene, ethylene or prop-2 yn- 1 -ylene) optionally substituted with halo; 10 1.58 Formula I or 1.57, wherein X is methylene or ethylene optionally substituted with halo; 1.59 Formula I, 1.57 or 1.58, wherein X is methylene optionally substituted with halo; 1.60 Formula 1, 1.57, 1.58 or 1.59, wherein X is methylene substituted with 15 bromo; 1.61 Formula I, 1.57, 1.58, 1.59 or 1.60, wherein X is ethylene; 1.62 Formula I or any of 1.57-1.61, wherein X is C 1 prop-2-yn-I-ylene; 1.63 Formula I or any of 1.57-1.62, wherein Y is a single bond, arylene (e.g., phenylene) or heteroarylene (e.g., pyridylene) optionally substituted with 20 halo; 1.64 Formula I or any of 1.57-1.63, wherein Y is arylene (e.g., phenylene) optionally substituted with halo; 21 1.65 Formula I or any of 1.57-1.64 wherein Y is phenylene optionally substituted with fluoro at the 3 or 5-position of the phenylene ring; 1.66 Formula I or any of 1.57-1.65, wherein Y is phenylene; 1.67 Formula I or any of 1.57-1.63, wherein Y is heteroarylene (e.g., 5 pyridylene) optionally substituted with halo; 1.68 Formula I or any of 1.57-1.63, wherein Y is a single bond; 1.69 Formula I or any of 1.57-1.63, wherein Y is heteroaryl (e.g., pyrid-2-yl); 1.70 Formula I or any of 1.57-1.69, wherein Z is H, aryl (e.g., phenyl), heteroaryl (e.g., pyrid-2-yl), halo (e.g., F, Br, Cl), haloCAalkyl (e.g., 10 trifluoromethyl), -C(O)-R', -N(R2)(R 3 ), or C 3

.

7 cycloalkyl optionally containing at least one atom selected from a group consisting of N or 0 (e.g., cyclopentyl, cyclohexyl, tetrahydro-2H-pyran-4-yl, or morpholinyl), optionally substituted with halo; 1.71 Formula I or any of 1.57-1.70, wherein Z is C 3

.

7 cycloalkyl optionally 15 containing at least one atom selected from a group consisting of N or 0 (e.g., cyclopentyl, cyclohexyl, tetrahydro-2H-pyran-4-yl, morpholinyl); 1.72 Formula I or any of 1.57-1.71, wherein Z is cyclopentyl; 1.73 Formula I or any of 1.57-1.71, wherein Z is tetrahydro-2H-pyran-4-yl; 1.74 Formula I or any of 1.57-1.70, wherein Z is heteroaryl (e.g., pyrid-2-yl) 20 optionally substituted with halo; 1.75 Formula I or any of 1.57-1.70 or 1.74, wherein Z is pyrid-2-yl; 1.76 Formula I or any of 1.57-1.70 or 1.74, wherein Z is 5-fluoro-pyrid-2-yl or 6-fluoro-pyrid-2-yl; 1.77 Formula I or any of 1.57-1.70, wherein Z is haloC1 4 alkyl (e.g., 25 trifluoromethyl); 1.78 Formula I or any of 1.57-1.70 or 1.77, wherein Z is trifluoromethyl; 1.79 Formula I or any of 1.57-1.70, wherein Z is -C(0)-R' and k' is C 1 4alky (e.g., methyl) or haloCloalkyl (e.g., trifluoromethyl); 1.80 Formula I or any of 1.57-1.70 or 1.79, wherein Z is-C(O)-R' and R' is 30 methyl; 22 1.81 Formula I or any of 1.57-1.70 or 1.79, wherein Z is-C(O)-R' and R' is trifluoromethyl; 1.82 Formula I or any of 1.57-1.70, wherein Z is aryl (e.g., phenyl) optionally substituted with halo (e.g., fluoro); 5 1.83 Formula I or any of 1.57-1.70 or 1.82, wherein Z is phenyl; 1.84 Formula I or any of 1.57-1.70, wherein Z is -N(R 2

)(R

3 ); 1.85 Formula I or any of 1.57-1.70 or 1.84, wherein Z is -N(R 2

)(R

3 ), wherein

R

2 and R 3 are methyl; 1.86 any of the preceding formulae wherein the compound is selected from a 10 group consisting of: O HN \' N Br O HN N N N N N N N N N F o O H N 0 HN- / F F 0 F N O HN O HN N O N 23 NH HN N N N OF F NN N F 0F O HN-- O HN-- / N F NN N NNN,. F O HN O HN N N N Nj 2NN NN N 24 O HN \ O HN N N N N N N N F and F F 1.87 any of the preceding formulae wherein the compounds inhibit phosphodiesterase-mediated (e.g., PDE 1-mediated, especially PDE lB 5 mediated) hydrolysis of cGMP, e.g., with an IC 50 of less than 1 gM, preferably less than 25 nM in an immobilized-metal affinity particle reagent PDE assay, for example, as described in Example 15, in free, salt or prodrug form, including its enantiomers, diastereoisomers and racernates, provided that when X is an unsubstituted methylene, Y is phenylene or 10 heteroarylene, and Z is aryl, heteroaryl, haloalkyl or cycloalkyl, then Z is a substituted with at least one halo (e.g., fluoro, chloro, bromo) or alkyl (e.g., methyl, ethyl) group. [0010] Preferably, the compound of the present invention is: 25 O HN N N Br N F O HN 1- N N-\ F F F or 0OHN\2 N, N NN N N F 5 in free, salt or prodrug form. 10013] In another aspect of the invention, there is provided a Compound of Formula (X): 26 0 RR N N N N / N N R 5 n R2 R4 R3 Formula (X) wherein 5 (i) RI is H or C 1

.

6 alkyl (e.g., methyl); (ii) R4 is H or C 1

.

6 alkyl and R 2 and R 3 are, independently, H or C 2 . 6 alkyl optionally substituted with halo or hydroxyl (e.g., R 2 and

R

3 are both methyl, or R 2 is H and R 3 is ethyl, isopropyl or hydroxyethyl), aryl, heteroaryl, (optionally hetero)arylalkoxy, 10 or (optionally hetero)arylCi- 6 alkyl; or

R

2 is H and R 3 and R 4 together form a di-, tri- or tetramethylene bridge (pref. wherein the R 3 and R 4 together have the cis configuration, e.g., where the carbons carrying R3 and R 4 have 15 the R and S configurations, respectively); (iii) R 5 is a substituted heteroarylCI.

6 alkyl, e.g., substituted with C1. 6 haloalkyl;

R

5 is -D-E-F, wherein: D is CI.6alkylene (e.g., methylene, ethylene or prop-2-yn-l 20 ylene); E is a single bond, alkynylene (e.g., -C=C-), arylene (e.g., phenylene) or heteroarylene (e.g., pyridylene); 27 F is H, aryl (e.g., phenyl), heteroaryl (e.g., pyridyl, e.g., pyrid 2-yl), halo (e.g., F, Br, Cl), haloCI.

6 alkyl (e.g., trifluoromethyl), -C(O)-Ris, -N(R 16 )(R,7), or C3.7cycloalkyl optionally containing at least one atom selected from a 5 group consisting of N or 0 (e.g., cyclopentyl, cyclohexyl, tetrahydro-2H-pyran-4-yl, or morpholinyl); wherein D, E and F are independently and optionally substituted with one or more halo (e.g., F, Cl or Br), C 1 . 6 alkyl (e.g., methyl), haloCI.

6 alkyl (e.g., trifluoromethyl), 10 for example, Z is heteroaryl, e.g., pyridyl substituted with one or more halo (e.g., 6-fluoropyrid-2-yl, 5-fluoropyrid-2 yl, 6-fluoropyrid-2-yl, 3-fluoropyrid-2-yl, 4-fluoropyrid-2 yl, 4,6-dichloropyrid-2-yl), haloC 1

.

6 alkyl (e.g., 5 trifluoromethylpyrid-2-yl) or CI.

6 alkyl (e.g., 5-methylpyrid 15 2-yl), or Z is aryl, e.g., phenyl, substituted with one or more halo (e.g., 4-fluorophenyl); or

R

5 is attached to one of the nitrogens on the pyrazolo portion of Formula (X) and is a moiety of Formula A 20

R

12 R11 Re Y R10 R9 Formula A wherein X, Y and Z are, independently, N or C, and R 8 , R 9 , RI and R 12 are independently H or halogen (e.g., Cl or F), and Rio 25 is halogen, alkyl, cycloalkyl, haloalkyl (e.g., trifluoromethyl), 28 aryl (e.g., phenyl), heteroaryl (e.g., pyridyl (for example pyrid 2-yl), or thiadiazolyl (e.g., 1,2,3-thiadiazol-4-yl)), diazolyl, triazolyl, tetrazolyl, arylcarbonyl (e.g., benzoyl), alkylsulfonyl (e.g., methylsulfonyl), heteroarylcarbonyl, or alkoxycarbonyl; 5 provided that when X, Y, or Z is nitrogen, Rg, R 9 , or R 10 , respectively, is not present; and (iv) R6 is H, alkyl, aryl, heteroaryl, arylalkyl (e.g., benzyl), arylamino (e.g., phenylamino), heterarylamino, N,N-dialkylamino,

N,N

diarylamino, or N-aryl-N-(arylakyl)amino (e.g., N-phenyl-N 10 (1,1 '-biphen-4-ylmethyl)amino); or

R

6 is -N(R, 8 )(Rig) wherein R 18 and Rig are independently H, C 1 . 6 alky or aryl (e.g., phenyl) wherein said aryl is optionally substituted with one or more halo (e.g., fluorophenyl, e.g., 4 15 fluorophenyl) or hydroxy (e.g., hydroxyphenyl, e.g., 4 hydroxyphenyl or 2-hydroxyphenyl); (v) n = 0 or 1; (vi) when n=1, A is -C(R 3

R[

4 )-; (vii) wherein R 13 and R 14 , are, independently, H or C 1

-

6 alkyl, aryl, 20 heteroaryl, (optionally hetero)arylalkoxy or (optionally hetero)arylalkyl; (viii) R 15 is Ci.

6 alkyl, haloC 1 .6alkyl, -OH or -OC 1

.

6 alkyl (e.g., -OCH 3 ); (ix) R 16 and R 17 are independently H or Ca.

6 alkyl; in free, salt or prodrug form. 25 [00141 In a further embodiment of this aspect of the invention, the Compound of Formula (X) is: 29 0 HN N N N N OH in free, salt or prodrug from. 100151 More preferably, the Compound of the Invention is selected from formula 5 1.54, in free, salt or prodrug form. More preferably, the Compound of the Invention is selected from formula 1.55, in free, salt or prodrug form. [00161 If not otherwise specified or clear from context, the following terms herein have the following meanings: (a) "Alkyl" as used herein is a saturated or unsaturated hydrocarbon moiety, 10 preferably saturated, preferably having one to six carbon atoms, which may be linear or branched, and may be optionally mono-, di- or tri substituted, e.g., with halogen (e.g., chloro or fluoro), hydroxy, or carboxy. (b) "Cycloalkyl" as used herein is a saturated or unsaturated nonaromatic 15 hydrocarbon moiety, preferably saturated, preferably comprising three to nine carbon atoms, at least some of which form a nonaromatic mono- or bicyclic, or bridged cyclic structure, and which may be optionally substituted, e.g., with halogen (e.g., chloro or fluoro), hydroxy, or. carboxy. 20 (c) "Aryl" as used herein is a mono or bicyclic aromatic hydrocarbon, preferably phenyl, optionally substituted, e.g., with alkyl (e.g., methyl), halogen (e.g., chloro or fluoro), haloalkyl (e.g., trifluoromethyl), hydroxy, carboxy, or an additional aryl or heteroaryl (e.g., biphenyl or pyridylphenyl). 30 (d) "Heteroaryl" as used herein is an aromatic moiety wherein one or more of the atoms making up the aromatic ring is sulfur or nitrogen rather than carbon, e.g., pyridyl or thiadiazolyl, which may be optionally substituted, e.g., with alkyl, halogen, haloalkyl, hydroxy or carboxy. 5 (e) For ease of reference, the atoms on the pyrazolo-pyrimidine core of the Compounds of the Invention are numbered in accordance with the numbering depicted in Formula I, unless otherwise noted. (f) Wherein Y is phenylene, the numbering is as follows: X 2 3 z 5Z 10 (g) It is intended that wherein the substituents end in "ene", for example, alkylene, phenylene or arylalkylene, said substitutents are intended to bridge or be connected to two other substituents. Therefore, methylene is intended to be -CH 2 - and phenylene intended to be -C 6

H

4 - and arylalkylene is intended to be -C 6

H

4

-CH

2 - or - CH 2

-C

6

H

4 -. 15 100171 Compounds of the Invention may exist in free or salt form, e.g., as acid addition salts. In this specification unless otherwise indicated, language such as "Compounds of the Invention" is to be understood as embracing all the novel compounds disclosed herein, e.g., I - or 2-substituted (6aR*,9aS*)-3-(phenylamino)-5-6a,7,8,9,9a hexahydro-5-methyl-cyclopent[4,5]imidazo[1, 2 -ajpyrazolo[4,3-e]pyrimidin-4(JH or 211) 20 one compounds, preferably 1- or 2-substituted (6aR,9aS) (phenylamino)-5-6a,7,8,9,9a hexahydro-5-methyl-cyclopent[4,5]imidazo[1, 2 -a]pyrazolo[4,3-ejpyrimidin-4(JH or 2H) one , the Compounds of Formula Q, e.g., any of 1.1-1.56, Compound of Formula (X), or Formula 1, e.g., any of 1.57-1.87, any of these compounds in any form, for example free or acid addition salt form, or where the compounds contain acidic substituents, in base 25 addition salt form. The Compounds of the Invention are intended for use as pharmaceuticals, therefore pharmaceutically acceptable salts are preferred. Salts which are unsuitable for pharmaceutical uses may be useful, for example, for the isolation or 31 purification of free Compounds of the Invention or their pharmaceutically acceptable salts, are therefore also included. 100181 Compounds of the Invention may in some cases also exist in prodrug form. A prodrug form is compound which converts in the body to a Compound of the 5 Invention. For example when the Compounds of the Invention contain hydroxy or carboxy substituents, these substituents may form physiologically hydrolysable and acceptable esters. As used herein, "physiologically hydrolysable and acceptable ester" means esters of Compounds of the Invention which are hydrolysable under physiological conditions to yield acids (in the case of Compounds of the Invention which have hydroxy 10 substituents) or alcohols (in the case of Compounds of the Invention which have carboxy substituents) which are themselves physiologically tolerable at doses to be administered. As will be appreciated the term thus embraces conventional pharmaceutical prodrug forms. [00191 The invention also provides methods of making the Compounds of the 15 Invention and methods of using the Compounds of the Invention for treatment of diseases and disorders as set forth below (especially treatment of diseases characterized by reduced dopamine Dl receptor signaling activity, such as Parkinson's disease, Tourette's Syndrome, Autism, fragile X syndrome, ADHD, restless leg syndrome, depression, cognitive impairment of schizophrenia, narcolepsy and diseases that may be 20 alleviated by the enhancement of progesterone-signaling such as female sexual dysfunction). DETAILED DESCRIPTION OF THE INVENTION Methods of Making Compounds of the Invention 100201 The compounds of the Invention, e.g., a (6aR*,9aS*)-3-(phenylamino)-5 25 6a,7,8,9,9a-hexahydro-5-methyl-cyclopent[4,5]imidazo[1,2-a]pyrazolo[4,3-e]pyrimidin 4(JH or 2H)-one compounds, preferably 1- or 2-substituted (6aR,9aS) (phenylamino)-5 6a,7,8,9,9a-hexahydro-5-methyl-cyclopent[4,5]imidazo[1,2-a]pyrazolo[4,3-e]pyrimidin 4(IH or 21)-one, a Compound of formula Q or I, e.g., any of 1.1 -.56 or 1.5701.87, or a Compound of Formula (X), and their pharmaceutically acceptable salts may be made 30 using the methods as described and exemplified herein and by methods similar thereto and by methods known in the chemical art. Such methods include, but not limited to, 32 those described below. If not commercially available, starting materials for these processes may be made by procedures, which are selected from the chemical art using techniques which are similar or analogous to the synthesis of known compounds. All references cited herein are hereby incorporated in their entirety by reference. 5 (00211 The Compounds of the Invention include their enantiomers, diastereoisomers and racemates, as well as their polymorphs, hydrates, solvates and complexes. Some individual compounds within the scope of this invention may contain double bonds. Representations of double bonds in this invention are meant to include both the E and the Z isomer of the double bond. In addition, some compounds within the 10 scope of this invention may contain one or more asymmetric centers. This invention includes the use of any of the optically pure stereoisomers as well as any combination of stereoisomers. 10022] Melting points are uncorrected and (dec) indicates decomposition. Temperature are given in degrees Celsius (*C); unless otherwise stated, operations are 15 carried out at room or ambient temperature, that is, at a temperature in the range of 18-25 *C. Chromatography means flash chromatography on silica gel; thin layer chromatography (TLC) is carried out on silica gel plates. NMR data is in the delta values of major diagnostic protons, given in parts per million (ppm) relative to tetramethylsilane (TMS) as an internal standard. Conventional abbreviations for signal shape are used. 20 Coupling constants (J) are given in Hz. For mass spectra (MS), the lowest mass major ion is reported for molecules where isotope splitting results in multiple mass spectral peaks Solvent mixture compositions are given as volume percentages or volume ratios. In cases where the NMR spectra are complex, only diagnostic signals are reported. [00231 Terms and abbreviations: 25 BuLi = n-butyllithium Bu'OH = tert-butyl alcohol, CAN = ammonium cerium (IV) nitrate, DIPEA = diisopropylethylamine, DMF = N,N-dimethylforamide, 30 DMSO = dimethyl sulfoxide, Et 2 O = diethyl ether, 33 EtOAc = ethyl acetate, equiv. = equivalent(s), h = hour(s), HPLC =high performance liquid chromatography, 5 LDA = lithium diisopropylamide MeOH = methanol, NBS = N-bromosuccinimide NCS = N-chlorosuccinimide NaHCO 3 = sodium bicarbonate, 10

NH

4 0H = ammonium hydroxide, Pd 2 (dba) 3 = tris[dibenzylideneacetone]dipalladium(0) PMB = p-methoxybenzyl, POC1 3 = phosphorous oxychloride, SOC1 2 = thionyl chloride, i5 TFA = trifluoroacetic acid, THF = tetrahedrofuran. [00241 The synthetic methods in this invention are illustrated below. The significances for the R groups are as set forth above for formula I unless otherwise 20 indicated. [0025] In an aspect of the invention, intermediate compounds of formula lIb can be synthesized by reacting a compound of formula Ila with a dicarboxylic acid, acetic anhydride and acetic acid mixing with heat for about 3 hours and then cooled: 0 HN NH 2 + HO O + (CH 3

CO)

2 0 9cOH N O 0 OH 900H~ 25 (Ila) (11b) wherein R' is methyl. 34 [0026] Intermediate Ic can be prepared by for example reacting a compound of Ilb with for example a chlorinating compound such as POCl 3 , sometimes with small amounts of water and heated for about 4 hours and then cooled 0 0 0 "C, dropwise N O N O + POCla + H 2 0 then 80 *C, - 4 h 0 N CI H H (llb) (lic) 5 [0027] Intermediate HId may be formed by reacting a compound of Ic with for example a P'-L in a solvent such as DMF and a base such as K 2 C0 3 at room temperature or with heating: O 0 REN K 2

CO

3 REN R I + P 1 -L K2I R O N Cl DMF ON CI H Pi 10 (lIc) (lid) wherein Pl is a protective group [e.g.,p-methoxybenzyl group (PMB)]; L is a leaving group such as a halogen, mesylate, or tosylate. 100281 Intermediate Ie may be prepared by reacting a compound of Ild with 15 hydrazine or hydrazine hydrate in a solvent such as methanol and refluxed for about 4 hours and then cooled: 0 0 RNMeOH R 1 . R + N H 2 N H 2 'H 2 0 _ _ _ _ _, N O N C1 reflux, 4h OlN N,

P

1 p1 H (lid) (Ilie) 35 [00291 Intermediate I1f can be synthesized by reacting a compound of I1c with hydrazine or hydrazine hydrate in a solvent such as methoxyethanol and refluxed for about 30 min and then cooled: o 0 R 1 MeOCH2CH2OH R N N | + NH 2

NH

2 N O N Cl reflux, 30 min O' N N'NH2 H H H (lic) (lf) 5 [00301 Intermediate Hg (wherein R' 3 phenyl), can be synthesized by reacting a compound of Ile with for example an aryl isothiocyanate or isocyanate in a solvent such as DMF and heated at I 10 *C for about 2 days and then cooled: o O HN-R 4 R' H N + R4-N=C=S DMF R1 N NH 0 N .NH 110 OC O"N ~ p1 N' 1 (lie) (lig) 10 100311 Intermediate ih may be synthesized from a compound of 1ig by removing the protective group P1 with an appropriate method. For example, if PI is a p methoxybenzyl group, then it can be removed with AIC1 3 at room temperature or with TFA under heated conditions. o HN-R 4 0 HN-R 4 'N deprotection R , 0 NNH N , NH p 1 t 15 (0g) (I1h) Intermediate Ih may also be prepared directly from a compound of Hf using the similar methods, but the yields are relatively low. 36 0 0 HN-R 4 R N

R

4 -N=C=O or R4-N=C=S

R

1 ,N N ,NH2 N -N O I N$ DMF, heating O N (Ilf) (1Ih) 100321 Intermediate II-I can be prepared by for example reacting a compound of IIh with for example a chlorinating compound such as POCl 3 .The reaction may be 5 carried out at atmospheric pressure and refluxed for about 2 days, or heated at 150-200 *C for about 10 min in a sealed vial with a microwave instrument. O HN-R 13 0 HN-R 1 3 R!,N reflux, 48h R, N NNH + or 200 *C, 5-10 min Cl'N N blH H sealed vial in microwave (1lh) (Il 10 [0033] Intermediate IIJ can be prepared by reacting a compound of II-I with an amino alcohol, e.g., (IR,2R)-1-amino-2-cyclopentanol, in a solvent such as DMF. The reaction may be heated overnight and then cooled. The reaction mixture may be purified by chromatography to obtain compound IIJ: 0

HN-R

4 o HN-R 4

H

2 N OH DMF, 1100C Ri.N NH N NH + HN iN N Cl N N OH (11-1) (LiJ) 15 [0034] Intermediate IIK can be formed by reacting a compound of IIJ with for example a dehydrating/halogenating agent such as SOC1 2 in a solvent such as CH 2

CI

2 at room temperature overnight or heated at 35 *C for about 4 hours, and then cooled to obtain cyclized compound (IIK). 37 0 HN-R 4 1 HN-R 4 R' SOC1 2 N - NH NH. HN N N'

CH

2

C

2 or MeOH, r.t. N N .*OH (IIJ) (IlK) 100351 Compound Ia and lb may be formed by reacting a compound of Ilk with for example a Z-Y-X-L in a solvent such as DMF and a base such as K 2 C0 3 at room 5 temperature or with heating: o HN-R 4 Z-Y-X-L 0 HN-R 4 0 HN-R 4 NH K 2 C:, R >N-X + RN N N DN MF NN N -Z N5 N b 0 'z (1K) (a)(b) wherein all the substitutents are as defined previously; L is a leaving group such as a 10 halogen, mesylate, or tosylate. 100361 X-Y-Z may also be introduced earlier by for example reacting IIg with Z Y-X-L and then perform similar procedure as described above to form compound Ia and Ib, as long as Z-Y-X will not be cleaved off in the P' deprotection step. 15 100371 The third synthetic route is also developed for the preparation of Compound Ia. [0038] Intermediate IVa may be formed by for example reacting a compound of Ie with POC1 3 and DMF: 38 0 0 RN 0 Cto r.t. R 1, N'H2 + POC1 3 + DMF N NH 0 N N'H 0 N N' H l 1 (lie) (Iva) wherein R' is methyl. 10039] Intermediate IVb may be formed by reacting a compound of IVa with for example a Z-Y-X-L in a solvent such as DMF and a base such as K 2

CO

3 at room 5 temperature or with heating: 0 0 O~ NH + Z-Y-X-L K2CO3 R N-X p1 1 (IVa) (IVb) [00401 Intermediate IVe may be synthesized from a compound of IVb by 10 removing the protective group P' with an appropriate method. For example, if P' is a PMB group, then it can be removed with CAN at room temperature: R 0 O RN

-

deprotection RN O N N' 'Y-Z O N--- 1 yH (lVb) (IVc) [00411 Intermediate IVd can be prepared by reacting a compound of IVc with for 15 example a chlorinating compound such as POCl 3 and refluxed for about 2 days, or heated at 150~200 *C for about 10 min in a sealed vial with a microwave instrument and then cooled: 39 0 O R N reflux, 48h R . N-X + POdl 3 O N N' Y-or 200 *C, 5-10 min N X H sealed vial in microwave Cl (IVc) (lVd) [0042] Intermediate IVe can be formed by reacting a compound of IVd with an 5 amino alcohol under basic condition in a solvent such as DMF and heated overnight then cooled: 0 0 R,.. RN O H 2 N OHRN N-X N +H baseHN N N Y-Z CI> N N NX - HNIA -X 'Y-Z DMF, 110 0 C -1OH (lVd) (IVe) 10 100431 Compound IVf may be formed by reacting a compound of IVe with for example a dehydrating/halogenating agent such as SOC1 2 in a solvent such as CH 2 Cl 2 at room temperature overnight or heated at 35 *C for about 4 hours, and then cooled. 0 0 SN-X SOC1 2 RN N-X N'Y-Z

Y

HN N y-Z CH 2 Cl 2 or MeOH, r.t. N N N Y-Z (lVe) (IVt) 15 100441 Compound IVg may be formed by reacting a compound of IVf with for example a halogenating agent such as NCS and a base such as LDA in a solvent such as THF at low temperature for several hours. 40 0 0 x R'. -N R) N-X NCS, NBS or 1 2 RkN N-X N N N Y-Z LDA or n-BuLi N N N Y-Z (IVOft 100451 Compound la may be formed by reacting a compound of IVg with for example an amine such as aniline in the present of a catalyst such as Pd 2 (dba) 3 in a 5 solvent such as dioxane at 100 *C overnight. R'' aniline N ' N N N-X cat., heating N N N Y-Z 0 b (IVg) (la) 100461 The invention thus provides methods of making Compounds of Formula 1, for example, comprising 10 (i) reacting a ( 6 aR, 9 aS)- 3 -(phenylamino)-5-6a,7,8,9,9a-hexahydro-5-methyl cyclopent[4,5]imidazo[1,2-alpyrazolo[4,3-e]pyrimidin-4(JH or 211)-one with a compound of formula Z-Y-X-L wherein L is a leaving group, e.g., halogen, mesylate, or tosylate, X, Y and Z are as defined above in Formula I, e.g., under basic conditions, for example wherein the (6aR,9aS)-3-(phenylamino)-5 15 6a,7,8,9,9a-hexahydro-5-methyl-cyclopent[4,5]imidazo[1,2-a]pyrazolo(4,3 e]pyrimidin-4(2H)-one is a compound of Formula IIK: 41 0 HN 0 Nl ' NH (IIK); or (ii) cyclization of a compound of Formula V R O HN NH HN N X-Y YOH Z 5 wherein, X, Y and Z are as defined above with reference to Formula 1, e.g., using a dehydrating/halogentating agent, for example thionyl chloride; and isolating the Compound of the Invention thus obtained. [00471 Similarly, the invention also provides methods of making Compounds of 10 Formula Q, for example, comprising: (i) reacting a (6aR,9aS)-3-(phenylamino)-5-6a,7,8,9,9a-hexahydro-5-methyl cyclopent[4,5]imidazo[ I,2-a]pyrazolo[4,3-ejpyrimidin-4(IH or 2H)-one with a compound of formula Z-Y-X-L wherein L is a leaving group, e.g., halogen, mesylate, or tosylate, X, Y and Z are as defined above in Formula Q, e.g., 15 under basic conditions, for example wherein the (6aR,9aS)-3-(phenylamino)-5 6a,7,8,9,9a-hexahydro-5-methyl-cyclopent[4,5]imidazo[1,2-a]pyrazolo[4,3 e]pyrimidin-4(2H)-one is a compound of Formula QK: 42 R4 O 'N.- R NH Nv N (QK); or (ii) cyclization of a compound of Formula QV R 4 0 AN-R 5 R

N.---

NH HN NN N X 5 wherein, X, Y and Z are as defined above with reference to Formula Q, e.g., using a dehydrating/halogentating agent, for example thionyl chloride. 43 [00481 Methods of using Compounds of the Invention 10049] The Compounds of the Invention are useful in the treatment of diseases characterized by disruption of or damage to cAMP and cGMP mediated pathways, e.g., as a result of increased expression of PDE 1 or decreased expression of cAMP and cGMP 5 due to inhibition or reduced levels of inducers of cyclic nucleotide synthesis, such as dopamine and nitric oxide (NO). By preventing the degradation of cAMP and cGMP by PDE1B, thereby increasing intracellular levels of cAMP and cGMP, the Compounds of the Invention potentiate the activity of cyclic nucleotide synthesis inducers. 10 [00501 The invention provides methods of treatment of any one or more of the following conditions: (i) Neurodegenerative diseases, including Parkinson's disease, restless leg, tremors, dyskinesias, Huntington's disease, Alzheimer's disease, and drug-induced movement disorders; 15 (ii) Mental disorders, including depression, attention deficit disorder, attention deficit hyperactivity disorder, bipolar illness, anxiety, sleep disorders, e.g., narcolepsy, cognitive'impairment, dementia, Tourette's syndrome, autism, fragile X syndrome, psychostimulant withdrawal, and drug addiction; 20 (iii) Circulatory and cardiovascular disorders, including cerebrovascular disease, stroke, congestive heart disease, hypertension, pulmonary hypertension, and sexual dysfunction; (iv) Respiratory and inflammatory disorders, including asthma, chronic obstructive pulmonary disease, and allergic rhinitis, as well as 25 autoimmune and inflammatory diseases; (v) Any disease or condition characterized by low levels of cAMP and/or cGMP (or inhibition of cAMP and/or cGMP signaling pathways) in cells expressing PDEl; and/or (vi) Any disease or condition characterized by reduced dopamine Dl 30 receptor signaling activity, 44 comprising administering an effective amount of a Compound of the Invention, e.g., a compound according to any of Formula I or 1.57-1.87, to a human or animal patient in need thereof. 100511 Similarly, the invention provides methods of treatment of any one or 5 more of the following conditions: (i) Neurodegenerative diseases, including Parkinson's disease, restless leg, tremors, dyskinesias, Huntington's disease, Alzheimer's disease, and drug-induced movement disorders; (ii) Mental disorders, including depression, attention deficit disorder, 10 attention deficit hyperactivity disorder, bipolar illness, anxiety, sleep disorders, e.g., narcolepsy, cognitive impairment, dementia, Tourette's syndrome, autism, fragile X syndrome, psychostimulant withdrawal, and drug addiction; (iii) Circulatory and cardiovascular disorders, including cerebrovascular 15 disease, stroke, congestive heart disease, hypertension, pulmonary hypertension, and sexual dysfunction; (iv) Respiratory and inflammatory disorders, including asthma, chronic obstructive pulmonary disease, and allergic rhinitis, as well as autoimmune and inflammatory diseases; 20 (v) Any disease or condition characterized by low levels of cAMP and/or cGMP (or inhibition of cAMP and/or cGMP signaling pathways) in cells expressing PDE I; and/or (vi) Any disease or condition characterized by reduced dopamine DI receptor signaling activity, 25 comprising administering an effective amount of a Compound of the Invention, e.g., a compound according to any of Formula Q or 1.1-1.56, or Formula (X), to a human or animal patient in need thereof. 100521 In an especially preferred embodiment, the invention provides methods of 30 treatment or prophylaxis for narcolepsy. In this embodiment, PDE I Inhibitors may be used as a sole therapeutic agent, but may also be used in combination or for co 45 administration with other active agents. Thus, the invention further comprises a method of treating narcolepsy comprising administering simultaneously, sequentially, or contemporaneously administering therapeutically effective amounts of (i) a PDE 1 Inhibitor, e.g., a compound according to any of Formula I or 1.57 5 1.87, and (ii) a compound to promote wakefulness or regulate sleep, e.g., selected from (a) central nervous system stimulants-amphetamines and amphetamine like compounds, e.g., methylphenidate, dextroamphetamine, methamphetamine, and pemoline; (b) modafinil, (c) antidepressants, e.g., tricyclics (including 10 imipramine, desipramine, clomipramine, and protriptyline) and selective serotonin reuptake inhibitors (including fluoxetine and sertraline); and/or (d) gamma hydroxybutyrate (GHB). in free or pharmaceutically acceptable salt form, to a human or animal patient in need thereof. 15 10053] The invention also comprises a method of treating narcolepsy comprising administering simultaneously, sequentially, or contemporaneously administering therapeutically effective amounts of (i) a PDE I Inhibitor, e.g., a compound according to any of Formula Q or 1.1 20 1.56, or Formula (X) and (ii) a compound to promote wakefulness or regulate sleep, e.g., selected from (a) central nervous system stimulants-amphetamines and amphetamine like compounds, e.g., methylphenidate, dextroamphetamine, methamphetamine, and pemoline; (b) modafinil, (c) antidepressants, e.g., tricyclics (including 25 imipramine, desipramine, clomipramine, and protriptyline) and selective serotonin reuptake inhibitors (including fluoxetine and sertraline); and/or (d) gamma hydroxybutyrate (GHB). in free or pharmaceutically acceptable salt form, to a human or animal patient in need thereof 30 100541 In another embodiment, the invention provides methods of treatment or 46 prophylaxis of a condition which may be alleviated by the enhancement of the progesterone signaling comprising administering an effective amount of a Compound of the Invention, e.g., a compound according to any of Formula 1.57-1.87 or Formula I, to a human or animal patient in need thereof. In still another embodiment, the invention 5 further provides methods of treatment or prophylaxis of a condition which may be alleviated by the enhancement of the progesterone signaling comprising administering an effective amount of a Compound of the Invention, e.g., a compound according to any of Formula Q or 1.1-1.56, or Formula (X) to a human or animal patient in need thereof. Disease or condition that may be ameliorated by enhancement of progesterone signaling 10 include, but are not limited to, female sexual dysfunction, secondary amenorrhea (e.g., exercise amenorrhoea, anovulation, menopause, menopausal symptoms, hypothyroidism), pre-menstrual syndrome, premature labor, infertility, for example infertility due to repeated miscarriage, irregular menstrual cycles, abnormal uterine bleeding, osteoporosis, autoimmmune disease, multiple sclerosis, prostate enlargement, 15 prostate cancer, and hypothyroidism. For example, by enhancing progesterone signaling, the PDE I inhibitors may be used to encourage egg implantation through effects on the lining of uterus, and to help maintain pregnancy in women who are prone to miscarriage due to immune response to pregnancy or low progesterone function. The novel PDE I inhibitors, e.g., as described herein, may also be useful to enhance the effectiveness of 20 hormone replacement therapy, e.g., administered in combination with estrogen/estradiol/estriol and/or progesterone/progestins in postmenopausal women, and estrogen-induced endometrial hyperplasia and carcinoma. The methods of the invention are also useful for animal breeding, for example to induce sexual receptivity and/or estrus in a nonhuman female mammal to be bred. 25 100551 In this embodiment, PDE I Inhibitors may be used in the foregoing methods of treatment or prophylaxis as a sole therapeutic agent, but may also be used in combination or for co-administration with other active agents, for example in conjunction with hormone replacement therapy. Thus, the invention further comprises a method of 30 treating disorders that may be ameliorated by enhancement of progesterone signaling comprising administering simultaneously, sequentially, or contemporaneously 47 administering therapeutically effective amounts of (i) a PDE I Inhibitor, e.g., a compound according to any of Formula 1.57-1.87 or Formula I, and (ii) a hormone, e.g., selected from estrogen and estrogen analogues (e.g., 5 estradiol, estriol, estradiol esters) and progesterone and progesterone analogues (e.g., progestins) in free or pharmaceutically acceptable salt form, to a human or animal patient in need thereof. 100561 The invention also comprises a method of treating disorders that may be 10 ameliorated by enhancement of progesterone signaling comprising administering simultaneously, sequentially, or contemporaneously administering therapeutically effective amounts of (i) a PDE I Inhibitor, e.g., a compound according to any of Formula Q, e.g., 1.1 1.56, or Formula (X) and 15 (ii) a hormone, e.g., selected from estrogen and estrogen analogues (e.g., estradiol, estriol, estradiol esters) and progesterone and progesterone analogues (e.g., progestins) in free or pharmaceutically acceptable salt form, to a human or animal patient in need thereof. 20 [00571 The invention also provides a method for enhancing or potentiating dopamine Dl intracellular signaling activity in a cell or tissue comprising contacting said cell or tissue with an amount of a Compound of the Invention, e.g., Formula Q, I, e.g., 1.1-1.56 or 1.57-1.87 , or Formula (X), sufficient to inhibit PDE I B activity. 25 100581 The invention also provides a method for enhancing or potentiating progesterone signaling activity in a cell or tissue comprising contacting said cell or tissue with an amount of a Compound of the Invention, e.g., Formula Q, I, e.g., 1.1-1.56 or 1.57-1.87 , or Formula (X), sufficient to inhibit PDE I B activity. 30 [0059 The invention also provides a method for treating a PDE 1-related, especially PDElB-related disorder, a dopamine Dl receptor intracellular signaling pathway 48 disorder, or disorders that may be alleviated by the enhancement of the progesterone signaling pathway in a patient in need thereof comprising administering to the patient an effective amount of a Compound of the Invention, e.g., Formula Q, I, e.g., 1.1-1.56 or 1.57-1.87 , or Formula (X), that inhibits PDEI B, wherein PDEIB activity modulates 5 phosphorylation of DARPP-32 and/or the GluRI AMPA receptor. [00601 The present invention also provides (i) a Compound of the Invention, e.g., Formula Q, I, e.g., 1.1-1.56 or 1.57-1.87 , or Formula (X), for use as a pharmaceutical, for example 10 for use in any method or in the treatment of any disease or condition as hereinbefore set forth, (ii) the use of a Compound of the Invention, e.g., Formula Q, 1, e.g., 1.1 1.56 or 1.57-1.87, or Formula (X), in the manufacture of a medicament for treating any disease or condition as hereinbefore set forth, 1 5 (iii) a pharmaceutical composition comprising a Compound of the Invention, e.g., Formula Q, I, e.g., 1.1-1.56 or 1.57-1.87 or Formula (X), in combination or association with a pharmaceutically acceptable diluent or carrier, and (iv) a pharmaceutical composition comprising a Compound of the 20 Invention, e.g., Formula Q, I, e.g., 1.1-1.56 or 1.57-1.87, or Formula (X), in combination or association with a pharmaceutically acceptable diluent or carrier for use in the treatment of any disease or condition as hereinbefore set forth. 25 100611 Therefore, the invention provides use of a Compound of the Invention, e.g., Formula Q, I, e.g., 1.1-1.56 or 1.57-1.87, or Formula (X), in free or pharmaceutically acceptable salt or prodrug form, or a Compound of the Invention in a pharmaceutical composition form, for the manufacture of a medicament for the treatment or prophylactic treatment of the following diseases: Parkinson's disease, restless leg, tremors, 30 dyskinesias, Huntington's disease, Alzheimer's disease, and drug-induced movement disorders; depression, attention deficit disorder, attention deficit hyperactivity disorder, 49 bipolar illness, anxiety, sleep disorder, narcolepsy, cognitive impairment, dementia, Tourette's syndrome, autism, fragile X syndrome, psychostimulant withdrawal, and/or drug addiction; cerebrovascular disease, stroke, congestive heart disease, hypertension, pulmonary hypertension, and/or sexual dysfunction; asthma, chronic obstructive 5 pulmonary disease, and/or allergic rhinitis, as well as autoimmune and inflammatory diseases; and/or female sexual dysfunction, exercise amenorrhoea, anovulation, menopause, menopausal symptoms, hypothyroidism, pre-menstrual syndrome, premature labor, infertility, irregular menstrual cycles, abnormal uterine bleeding, osteoporosis, multiple sclerosis, prostate enlargement, prostate cancer, hypothyroidism, estrogen 10 induced endometrial hyperplasia or carcinoma; and/or any disease or condition characterized by low levels of cAMP and/or cGMP (or inhibition of cAMP and/or cGMP signaling pathways) in cells expressing PDEI, and/or by reduced dopamine Dl receptor signaling activity; and/or any disease or condition that may be ameliorated by the enhancement of progesterone signaling; comprising administering an effective amount of 15 a Compound of the Invention, or a pharmaceutical composition comprising a Compound of the Invention, , e.g., Formula Q, I, e.g., 1.1-1.56 or 1.57-1.87, or Formula (X), to a patient in need of such treatment. [00621 The words "treatment" and "treating" are to be understood accordingly as 20 embracing prophylaxis and treatment or amelioration of symptoms of disease as well as treatment of the cause of the disease 100631 Compounds of the Invention are in particular useful for the treatment of Parkinson's disease, narcolepsy and female sexual dysfunction. 25 100641 Compounds of the Invention, e.g., Formula Q, I, e.g., 1.1-1.56 or 1.57 1.87, or Formula (X), may be used as a sole therapeutic agent, but may also be used in combination or for co-administration with other active agents. For example, as Compounds of the Invention potentiate the activity of DI agonists, such as dopamine, 30 they may be simultaneously, sequentially, or contemporaneously administered with conventional dopaminergic medications, such as levodopa and levodopa adjuncts 50 (carbidopa, COMT inhibitors, MAO-B inhibitors), dopamine agonists, and anticholinergics, e.g., in the treatment of a patient having Parkinson's disease. In addition, the novel PDE 1 inhibitors, e.g., as described herein, may also be administered in combination with estrogen/estradiol/estriol and/or progesterone/progestins to enhance 5 the effectiveness of hormone replacement therapy or treatment of estrogen-induced endometrial hyperplasia or carcinoma. [0065] Dosages employed in practicing the present invention will of course vary depending, e.g. on the particular disease or condition to be treated, the particular 10 Compound of the Invention used, the mode of administration, and the therapy desired. Compounds of the Invention may be administered by any suitable route, including orally, parenterally, transdermally, or by inhalation, but are preferably administered orally. In general, satisfactory results, e.g. for the treatment of diseases as hereinbefore set forth are indicated to be obtained on oral administration at dosages of the order from about 0.0 1 to 15 2.0 mg/kg. In larger mammals, for example humans, an indicated daily dosage for oral administration will accordingly be in the range of from about 0.75 to 150 mg, conveniently administered once, or in divided doses 2 to 4 times, daily or in sustained release form. Unit dosage forms for oral administration thus for example may comprise from about 0.2 to 75 or 150 mg, e.g. from about 0.2 or 2.0 to 50, 75 or 100 mg of a 20 Compound of the Invention, together with a pharmaceutically acceptable diluent or carrier therefor. 10066] Pharmaceutical compositions comprising Compounds of the Invention may be prepared using conventional diluents or excipients and techniques known in the galenic art. Thus oral dosage forms may include tablets, capsules, solutions, suspensions 25 and the like. EXAMPLES 30 EXAMPLE 1 51 (6aR,9aS)-5,6a,7,8,9,9a-hexahydro-5-methyl-3-(phenylamino)-2-((tetrahydro-2H pyran-4-yl)methyl)-cyclopent[4,5] imidazo[ 1,2-al pyrazolo[4,3-e pyrimidin-4(2H)-one ~JN NN N 5 [00671 (a) 7-(4-Methoxybenzyl)-5-methyl-3-(phenylamino)-IH-pyrazolo[3,4 d]pyrimidine-4,6(5H,7H)-dione Phenyl isothiocyanate (3.9 mL, 32.7 mmol) is added to a suspension of 6 hydrazinyl- I -(4-methoxybenzyl)-3-methylpyrimidine-2,4(1 H,3 H)-dione (0.45 g, 1.6 10 mmol) in DMF (12 mL). The reaction mixture is heated at 120 *C for 40 hours, and then evaporated to remove solvent under reduced pressure. The residue is washed with hexanes, and then treated with MeOH (125 mL), and stored at -15 *C for 2 days to give a crystalline solid. The solid is recrystallized from CH 3 0H-EtOAc to afford 2.5 g of product (Yield: 61%). 'H NMR (400 MHz, DMSO-d 6 ) 13.21 (s, 3H), 3.73 (s, 3H), 5.01 15 (s, 2H), 6.88-7.36 (m, 9H). MS (FAB) m/z 378.3 [M+H]*. [00681 (b) 5-Methyl-3-(phenylamino)-1H-pyrazolo[3,4-d]pyrimidine-4,6(5H,7H) dione To a solution of 7-(4-Methoxybenzyl)-5-methyl-3-(phenylamino)- 1 H 20 pyrazolo[3,4-d]pyrimidine-4,6(5H,7H)-dione (6.6 g, 17.5 mmol) in CH 2

C

2 (200 mL) is added TFA (30 mL) slowly at room temperature, followed by adding trifluoromethanesulfonic acid (10 mL) dropwise. After the reaction mixture is stirred at r.t. for 2 hours, solvent is removed under reduced pressure, and then 200 mL of IN NaOH is added with cooling. The mixture is extracted with ethyl acetate five times, dried 25 over Na 2

SO

4 , and then filtered. The filtrate is evaporated to dryness to give 4.3 g of crude product as white solids (Yield: 96%). MS (ESI) m/z 258.1 [M+H)*. 52 [0069] (c) 6-Chloro-5-methyl-3-(phenylamino)- I H-pyrazolo[3,4-d]pyrimidin 4(5H)-one 5-methyl-3-(phenylamino)- I H-pyrazolo[3,4-d]pyrimidine-4,6(5H,7H1)-dione (4.3 g, 16.7 mmol) is refluxed in POC1 3 (120 mL) for 2 days, and then POC1 3 is removed 5 under reduced pressure. The residue is suspended in 100 mL of water and carefully adjusted to pH = 1-2 with 7% NH 4 0H upon cooling. The mixture is then extracted with

CH

2 C1 2 and MeOH (10:1, v/v) five times. The combined organic phase is washed with water, dried over Na 2

SO

4 , and then evaporated under reduced vacuum to give 3.3 g of crude product (Yield: 72%). MS (ESI) m/z 276.1 [M+H]*. 10 [0070] (d) 6-((1 R,2R)-2-hydroxycyclopentylamino)-5-methyl-3-(phenylamino) 2H-pyrazolo[3,4-d]pyrimidin-4(5H)-one A solution of 6-chloro-5-methyl-3-(phenylamino)-I I-pyrazolo[3,4-d]pyrimidin 4(5H)-one (3.3 g, 12 mmol), (IR,2R)-2-aminocyclopentanol hydrochloride(2 g, 14.4 15 mmol) and DIPEA (4.6 mL, 27 mmol) in DMF (25 mL) is heated at 120 "C for 5 hours. Solvent is removed under reduced pressure. The residue is dissolved in CH 2 Cl 2 and MeOH (10:1, v/v), and then washed with water three times. The solution is dried over MgSO 4 and evaporated to dryness to give 3.5 g of crude product. MS (ESI) m/z 341.2 [M+H]+. 20 [00711 (e) (6aR,9aS)-5,6a,7,8,9,9a-hexahydro-5-methyl-3-(phenylamino) cyclopent[4,5]imidazo[ 1,2-a]pyrazolo[4,3-e]pyri midin-4(2H-)-one 2.0 M solution of thionyl chloride in CH 2

CI

2 (7.5 mL, 15.4 mmol) is added dropwise to a solution of 6-((l R,2R)-2-hydroxycyclopentylamino)-5-methyl-3 25 (phenylamino)-2H-pyrazolo[3,4-d]pyrimidin-4(5H)-one (3.5 g, 10.3 mmol) in CH 2 Cl2 (30 mL) at room temperature. After the completion of the addition, the reaction mixture is stirred at r.t. for 2h. Solvent and excess SOCl 2 are removed under reduced pressure. The residue is suspended in water (100 mL), and then carefully neutralized with 7% ammonium hydroxide (5 mL) to pH = 6.5 - 7. The mixture is extracted with CH 2 Cl 2 and 30 MeOH (10:1, v/v) five times. The combined organic phase is washed with water, dried 53 over Na 2

SO

4 , and then evaporated under reduced vacuum to give 2.9 g of crude product. MS (ESI) m/z 323.2 [M+H]*. 100721 (f) (6aR,9aS)-5,6a,7,8,9,9a-hexahydro-5-methyl-3-(phenylamino)-2 5 ((tetrahydro-2H-pyran-4-yl)methyl)-cyclopent[4,5]imidazo[ 1,2-a]pyrazolo[4,3 e]pyrimidin-4(2H)-one A mixture of (6aR,9aS)-5,6a,7,8,9,9a-hexahydro-5-methyl-3-(phenylamino) cyclopent[4,5]imidazo[1, 2 -alpyrazolo[4,3-e]pyrimidin-4(2H)-one (50 mg, 0.155 mmol), 4-(iodomethyl)-tetrahydro-2H-pyran (70 mg, 0.310 mmol), and Cs 2

CO

3 (10 1 mg, 0.310 10 mmol) in DMF (I mL) is heated in microwave at 140 "C for 30 min. After cooling, the mixture is filtered through a 0.45 pm microfilter, and the filtrate is purified by a semi preparative HPLC to give pure product as white powder. MS (ESI) m/z 421.2 [M+H]*. EXAMPLE 2 15 ( 6 aR, 9 aS)-5,6a,7,8,9,9a-hexahydro-5-methyl-3-(phenylamino)-1-((tetrahydro-2H pyran- 4 -yl)methyl)-cyclopent[4,5]imidazo[1,2-alpyrazolo[4,3-e]pyrimidin-4(1H)-one 0 HN- NN N (R ) (s) 20 100731 A mixture of (6aR,9aS)-5,6a,7,8,9,9a-hexahydro-5-methyl-3 (phenylamino)-cyclopent[4,5]imidazo[I, 2 -a]pyrazolo[4,3-e]pyrimidin-4(2H)-one (50 mg, 0.155 mmol), 4-(iodomethyl)-tetrahydro-2H-pyran (70 mg, 0.310 mmol), and Cs 2

CO

3 (101 mg, 0.310 mmol) in DMF (I mL) is heated in microwave at 140 *C for 30 min. After cooling, the mixture is filtered through a 0.45 pim microfilter, and the filtrate is 25 purified by a semi-preparative HPLC to give pure product as white powder. MS (ESI) m/z 421.2 [M+H]*. 54 EXAMPLE 3

(

6 aR, 9 aS)-5, 6 a,7,8,9,9a-hexahydro-5-methyl-3-(phenylamino)-1-(3-(dimethylamino) 2 -methylpropyl)-cyclopent[4,5]imidazoll,2-a pyrazolo[4,3-ejpyrimidin-4(lH)-one 5 O HN- G N N

N

100741 The synthesis method is analogous to example 2 wherein 3-chloro-N,N,2 trimethylpropan- 1-amine is added instead of 4 -(iodomethyl)-tetrahydro-2H-pyran. MS 10 (ESI) m/z 422.3 [M+H]*. EXAMPLE 4

(

6 aR, 9 aS)-5,6a,7,8,9,9a-hexahydro-5-methyl-3-(phenylamino)-1 (cyclopentylmethyl)-cyclopent[4,5]imidazoI1,2-a pyrazolo[4,3-el pyrimidin-4(lH) 15 one o HN N N 10075] The synthesis method is analogous to example 2 wherein cyclopentylmethyl iodide is added instead of 4 -(iodomethyl)-tetrahydro-2H-pyran. MS 20 (ESI) m/z 405.2 [M+H]*. 55 EXAMPLE 5 (6aR,9aS)-5,6a,7,8,9,9a-hexahydro-5-methyl-3-(phenylamino)-2 (cyclopentylmethyl)-cyclopent[4,5]imidazo[1,2-a pyrazolo[4,3-el pyrimidin-4(2H) one 5 0 HN N NN 100761 The synthesis method is analogous to example I wherein cyclopentylmethyl iodide is added instead of 4 -(iodomethyl)-tetrahydro-2H-pyran. MS 10 (ESI) m/z 405.2 [M+H]*. EXAMPLE 6

(

6 aR, 9 aS)-5, 6 a,7,8,9,9a-hxahydro-5-methyl-3-(phenylamino)-1 -(3-phenylprop-2 ynyl)-cyclopent[4,5]imidazo[l,2-a]pyrazolo[4,3-elpyrimidin-4(lH)-one 15 O HN- NN N [00771 The synthesis method is analogous to example 2 wherein (3-bromoprop-l ynyl)benzene is added instead of 4 -(iodomethyl)-tetrahydro-2H-pyran. MS (ESI) m/z 20 437.2 [M+H]*. EXAMPLE 7 56 (6aR,9aS)-S,6a,7,8,9,9a-hexahydro-5-methyl-3-(phenyla mino)-1 -(4-acetyiphenetbyl) cyclopentl4,5lim idazoll ,2-al pyrazolol4,3-el pyrimidin-4(I H)-one 0 100781 The synthesis method is analogous to example 2 wherein I -(4-(2 chloroethyl)phenyl)ethanone is added instead of 4-(iodomethyl)-tetrahydro-2H-pyran. MS (ESI) m/z 469.1 (M+H]f. 10 EXAMPLE 8 (6aR,9aS)-5,6a,7,8,9,9a- hcxa hyd ro-5- methyl-3-(p henylam ino)-2-(4-acety Iphen ethyl) cyclopentl4,5]imidazoll1,2-ajpyrazoloj4,3-ejpyrimidin-4(2H)-one 0 HNjii N 0 15 [00791 The synthesis method is analogous to example I wherein 1-(4-(2 chloroethyl)phenyl)ethanone is added instead of 4-(iodomethyl)-tetrahydro-2H-pyran. MS (ESI) m/z 469.2 [M+H]+. 20 EXAMPLE 9 (6aR,9aS)-S,6a,7,8,9,9a-hexa hyd ro-5-metliyl-3-(p heny lam ino)-1 -(3 -fluo ro-4-(2,2,2. trifluoroacetyI)benzyI)-cyclopent[4,51imidazol1 ,2-alpyrazolo 14,3-el pyrimidin-4(1H) one 57 o HN NN N 'N F F F F 0 100801 The synthesis method is analogous to example 2 wherein 1-(4 5 (bromomethyl)-2-fluorophenyl)-2,2,2-trifluoroethanone is added instead of 4 (iodomethyl)-tetrahydro-2H-pyran. MS (ESI) m/z 527.2 [M+H]*. EXAMPLE 10

(

6 aR, 9 aS)-5,6a,7,8,9,9a-hexahydro-5-methyl-3-(phenylamino)-2-(3-fluoro-4-(2,2,2 10 trifluoroacetyl)benzyl)-cyclopent[4,5]imidazo[i,2-alpyrazolol4,3-elpyrimidin-4(2H) one 0 HN- N N F F F 0 15 [00811 The synthesis method is analogous to example I wherein 1-(4 (bromomethyl)-2-fluorophenyl)-2,2,2-trifluoroethanone is added instead of 4 (iodomethyl)-tetrahydro-2H-pyran. MS (ESI) m/z 527.2 [M+H]. EXAMPLE 11 58

(

6 aR, 9 aS)-5,6a,7,8,9,9a-hexahydro-5-methyl-3-(phenylamino)-I -(4 (trifluoromethyl)phenethyl)-cyclopent[4,51imidazo I ,2-alpyrazolo[4,3-elpyrimidin 4 (1H)-one 0 HN N NN F 5 F F 100821 The synthesis method is analogous to example 2 wherein 1-(2 bromoethyl)-4-(trifluoromethyl)benzene is added instead of 4 -(iodomethyl)-tetrahydro 2H-pyran. MS (ESI) m/z 495.2 [M+H]*. 10 EXAMPLE 12

(

6 aR, 9 aS)-5, 6 a,7,8,9,9a-hexahydro-5-methyl-3-(phenylamino)-2-(4 (trifluoromethy1)phenethyl)-cyclopent[4,51imidazo1, 2 -aIpyrazolo[4,3-ejpyrimidin 4(2H)-one 15 0 HN N F 10083] The synthesis method is analogous to example I wherein I -(2 bromoethyl)-4-(trifluoromethyl)benzene is added instead of 4 -(iodomethyl)-tetrahydro 20 2H-pyran. MS (ESI) m/z 495.2 [M+H]*. 59 EXAMPLE 13

(

6 aR, 9 aS)-5,6a,7,8,9,9a-hexahydro-5-methyl-3-(phenylamino)-2-(bromo(4-(5 fluoropyridin-2-yl)phenyl)methyl)-cyclopent[4,5]imidazo[1,2-ajpyrazolo[4,3 5 ejpyrimidin-4(2H)-one 0 HN- NN Br N F [00841 A mixture of (6aR,9aS)-5,6a,7,8,9,9a-hexahydro-5-methyl-3 10 (phenylamino)-cyclopent[4,5]imidazo[ 1, 2 -alpyrazolo[4,3-e]pyrimidin-4(2H)-one (35 mg, 0.109 mmol), 2 -(4-(dibromomethyl)phenyl)-5-fluoropyridine, and K 2

CO

3 (15 mg, 0.109 mmol) in DMF (3 mL) is stirred at room temperature overnight. The mixture is filtered through a 0.45 prm microfilter, and the filtrate is purified by a semi-preparative HPLC to give product as white powder. MS (ESI) m/z 585.9 [M+H]*. 15 EXAMPLE 14 (6aR,9aS)-5,6a,7,8,9,9a-hexahydro-5-methyl-3-(phenylamino)-2-((4-(6 fluoropyridin-2-yl)phenyl)methyl)-cyclopent[4,5]imidazo[1,2-alpyrazolo[4,3 ejpyrimidin-4(2H)-one 20 60 0 HN\' _N NN N NIP F 100851 This compound may be made using similar method as in example 13 wherein 2-(4-(bromomethyl)phenyl)-6-fluoropyridine may be used instead of 2-(4 (dibromomethyl)phenyl)-5-fluoropyridine. 5 EXAMPLE 15 Measurement of PDE1B inhibition in vitro using IMAP Phosphodiesterase Assay Kit [00861 Phosphodiesterase IB (PDEI B) is a calcium/calmodulin dependent 10 phosphodiesterase enzyme that converts cyclic guanosine monophosphate (cGMP) to 5' guanosine monophosphate (5'-GMP). PDEIB can also convert a modified cGMP substrate, such as the fluorescent molecule cGMP-fluorescein, to the corresponding GMP-fluorescein. The generation of GMP-fluorescein from cGMP-fluorescein can be quantitated, using, for example, the IMAP (Molecular Devices, Sunnyvale, CA) 15 immobilized-metal affinity particle reagent. [00871 Briefly, the IMAP reagent binds with high affinity to the free 5'-phosphate that is found in GMP-fluorescein and not in cGMP-fluorescein. The resulting GMP fluorescein - IMAP complex is large relative to cGMP-fluorescein. Small fluorophores that are bound up in a large, slowly tumbling, complex can be distinguished from 20 unbound fluorophores, because the photons emitted as they fluoresce retain the same polarity as the photons used to excite the fluorescence. 100881 In the phosphodiesterase assay, cGMP-fluorescein, which cannot be bound to IMAP, and therefore retains little fluorescence polarization, is converted to GMP fluorescein, which, when bound to IMAP, yields a large increase in fluorescence 61 polarization (Amp). Inhibition of phosphodiesterase, therefore, is detected as a decrease in Amp. [0089] Enzyme assay Materials: All chemicals are available from Sigma-Aldrich (St. Louis, MO) except for 5 IMAP reagents (reaction buffer, binding buffer, FL-QMP and IMAP beads), which are available from Molecular Devices (Sunnyvale, CA). Assay: 3',5'-cyclic-nucleotide-specific bovine brain phosphodiesterase (Sigma, St. Louis, MO) is reconstituted with 50% glycerol to 2.5 U/ml. One unit of enzyme will hydrolyze 1.0 Vmole of 3',5'-cAMP to 5'-AMP per min at pH 7.5 at 30*C. One part 10 enzyme is added to 1999 parts reaction buffer (30 pM CaCI 2 , 10 U/ml of calmodulin (Sigma P2277), 10mM Tris-HCl pH 7.2, 10mM MgC 2 , 0.1% BSA, 0.05% NaN 3 ) to yield a final concentration of 1.25mU/ml. 99 pl of diluted enzyme solution is added into each well in a flat bottom 96-well polystyrene plate to which 1 pI of test compound dissolved in 100% DMSO is added. The compounds are mixed and pre-incubated with 15 the enzyme for 10 min at room temperature. 100901 The FL-GMP conversion reaction is initiated by combining 4 parts enzyme and inhibitor mix with I part substrate solution (0.225 pM) in a 384-well microtiter plate. The reaction is incubated in dark at room temperature for 15 min. The reaction is halted by addition of 60 pl of binding reagent (1:400 dilution of IMAP beads 20 in binding buffer supplemented with 1:1800 dilution of antifoam) to each well of the 384 well plate. The plate is incubated at room temperature for I hour to allow IMAP binding to proceed to completion, and then placed in an Envision multimode microplate reader (PerkinElmer, Shelton, CT) to measure the fluorescence polarization (Amp). 100911 A decrease in GMP concentration, measured as decreased Amp, is 25 indicative of inhibition of PDE activity. IC 50 values are determined by measuring enzyme activity in the presence of 8 to 16 concentrations of compound ranging from 0.0037 nM to 80,000 nM and then plotting drug concentration versus AmP, which allows

IC

5 0 values to be estimated using nonlinear regression software (XLFit; IDBS, Cambridge, MA). 30 [0092] The Compounds of the Invention are selected and tested in an assay as described or similarly described herein for PDE1 inhibitory activity, which compounds 62 generally have IC 50 values of less than IipM, e.g., the Compounds of formulae 1.54 and 1.55 generally have ICSO values of less than 250 nM. Example 16 5 PDE1 inhibitor effect on sexual response in female rats 10093] The effect of PDE1 inhibitors on Lordosis Response in female rats is measured as described in Mani, et al., Science (2000) 287: 1053. Ovariectomized and cannulated wild-type rats are primed with 2 pg estrogen followed 24 hours later by intracerebroventricular (icv) injection of progesterone (2 pig), PDEI inhibitors of the 10 present invention (0.1mg, 1.0mg or 2.5mg) or sesame oil vehicle (control). The rats are tested for lordosis response in the presence of male rats. Lordosis response is quantified by the lordosis quotient (LQ = number of lordosis/10 mounts x 100). It will be observed that the LQ for estrogen-primed female rats receiving compounds of the present invention, at 0.1 mg, will generally be similar to estrogen-primed rats receiving 15 progesterone and higher than for estrogen-primed rats receiving vehicle. 63

Claims (20)

1. A 1- or 2-substituted ( 6 aR*, 9 aS*)-3-(phenylamino)-5-6a,7,8,9,9a-hexahydro-5 5 methyl-cyclopent(4,5]imidazo[ 1, 2 -a]pyrazolo[4,3-e]pyrimidin-4(IH or 2H)-one, in free, salt or prodrug form.

2. The compound according to claim 1, wherein said compound is a Compound of formula Q R 4 O N.--R5 N 10 w Formula Q wherein (i) X is C 1 -alkylene (e.g., methylene, ethylene or prop-2-yn-1-ylene); (ii) Y is a single bond, alkynylene (e.g., -C=C-), arylene (e.g., phenylene) or 15 heteroarylene (e.g., pyridylene); (iii) Z is H, aryl (e.g., phenyl), heteroaryl (e.g., pyridyl, e.g., pyrid-2-yl), halo (e.g., F, Br, Cl), haloC 1 . 6 alkyl (e.g., trifluoromethyl), -C(O)-R', N(R2)(R ), or C3-7cycloalkyl optionally containing at least one atom selected from a group consisting of N or 0 (e.g., cyclopentyl, cyclohexyl, 20 tetrahydro-2H-pyran-4-yl, or morpholinyl); (iv) R' is C i. 6 alkyl, haloC 1 . 6 alkyl, -OH or -OC 1 . 6 alkyl (e.g., -OCH 3 ); (v) R 2 and R 3 are independently H or Ci. 6 alkyl; 64 (vi) R 4 and R 5 are independently H, CI 6 alky or aryl (e.g., phenyl) optionally substituted with one or more halo (e.g., fluorophenyl, e.g., 4 fluorophenyl), hydroxy (e.g., hydroxyphenyl, e.g., 4-hydroxyphenyl or 2 hydroxyphenyl) or CI- 6 alkoxy; 5 (vii) wherein X, Y and Z are independently and optionally substituted with one or more halo (e.g., F, Cl or Br), Ci. 6 alkyl (e.g., methyl), haloC. 6 alkyl (e.g., trifluoromethyl), for example, Z is heteroaryl, e.g., pyridyl substituted with one or more halo (e.g., 6-fluoropyrid-2-yl,

5-fluoropyrid 2 -yl, 6-fluoropyrid-2-yl, 3-fluoropyrid-2-yl, 4-fluoropyrid-2-yl, 4,6 10 dichloropyrid-2-yl), haloCI. 6 alkyl (e.g., 5-trifluoromethylpyrid-2-yl) or C1. 6 alkyl (e.g., 5-methylpyrid-2-yl), or Z is aryl, e.g., phenyl, substituted with one or more halo (e.g., 4-fluorophenyl), in free, salt or prodrug form, provided that when X is an unsubstituted methylene, Y is phenylene or heteroarylene, and Z is aryl, heteroaryl, haloalkyl or cycloalkyl, then Z is 15 substituted with at least one halo (e.g., fluoro, chloro, bromo) or alkyl (e.g., methyl, ethyl) group. 3. The compound according to claim 1 or 2, wherein said compound is a Compound of formula I 20 0 HN-O NN N ~ X Y' HN SN N X-Y z 65 Formula I wherein (i) X is C. 4 alkylene; (ii) Y is a single bond, alkynylene, arylene or heteroarylene; 5 (iii) Z is H, aryl, heteroaryl, halo, haloCI.4alkyl, -C(O)-R', -N(R 2 )(R 3 ), or C 3 . 7cycloalkyl optionally containing at least one atom selected from a group consisting of N or 0; (iv) R' is Ci.4alkyl, haloCI. 4 alkyl; (v) R2 and R 3 are independently H or Ci4alkyl, 10 (vi) wherein X, Y and Z are independently and optionally substituted with halo, in free, salt or prodrug form, including its enantiomers, diastereoisomers and racemates, provided that when X is an unsubstituted methylene, Y is phenylene or heteroarylene, and Z is aryl, heteroaryl, haloalkyl or cycloalkyl, then Z is a substituted with at least one 15 halo (e.g., fluoro, chloro, bromo) or alkyl (e.g., methyl, ethyl) group. 4. A compound according to claim 1, 2 or 3, wherein said compound is formula II: HN N- N QN N N y-Z 20 5. A compound according to any of claims 1-4, selected from the compounds of formula 1.53. 66

6. A compound according to any of claims 1-5, selected from the compounds of formula 1.54.

7. A compound according to any of claims 1-6, selected from the compounds of formula 5 1.55

8. A compound according to any of claims 1-4, selected from the compounds of formula 1.86. 10 9. A compound according to any of claims 1-5, wherein said compound is O HN \z Br N N N (R) (S) N F

10. A compound according to claim 1, wherein said compound is O HN (E (R) (S) F F O F 15

11. A compound according to claim 1, wherein said compound is 67 O HN \ N N F

12. A pharmaceutical composition comprising a compound according to any of claims 1 11 in admixture with a pharmaceutically acceptable diluent or carrier. 5 13. A method of treating any of the following conditions: Parkinson's disease, restless leg, tremors, dyskinesias, Huntington's disease, Alzheimer's disease, and drug induced movement disorders; depression, attention deficit disorder, attention deficit hyperactivity disorder, bipolar illness, anxiety, sleep disorder, narcolepsy, cognitive impairment, dementia, Tourette's syndrome, autism, fragile X syndrome, 10 psychostimulant withdrawal, and/or drug addiction; cerebrovascular disease, stroke, congestive heart disease, hypertension, pulmonary hypertension, and/or sexual dysfunction; asthma, chronic obstructive pulmonary disease, and/or allergic rhinitis, as well as autoimmune and inflammatory diseases; and/or female sexual dysfunction, exercise amenorrhoea, anovulation, menopause, menopausal symptoms, 15 hypothyroidism, pre-menstrual syndrome, premature labor, infertility, irregular menstrual cycles, abnormal uterine bleeding, osteoporosis, multiple sclerosis, prostate enlargement, prostate cancer, hypothyroidism, estrogen-induced endometrial hyperplasia or carcinoma; and/or any disease or condition characterized by low levels of cAMP and/or cGMP (or inhibition of cAMP and/or cGMP signaling pathways) in 20 cells expressing PDEI, and/or by reduced dopamine Dl receptor signaling activity; and/or any disease or condition that may be ameliorated by the enhancement of progesterone signaling; comprising administering an effective amount of a compound according to any of claims I-11, or a pharmaceutical composition according to claim 12, to a patient in need of such treatment. 68

14. The method of claim 13, wherein the condition is Parkinson's disease.

15. The method of claim 13, wherein the condition is cognitive impairment. 5

16. The method of claim 13, wherein the condition is narcolepsy.

17. The method of claim 16 further comprising administering a compound or compounds selected from central nervous system stimulants, modafinil, antidepressants, and 10 gamma hydroxybutyrate, to a patient in need thereof.

18. The method of claim 13, wherein said condition is female sexual dysfunction.

19. The method of claim 18, further comprising administering a compound or compounds 15 selected from a group consisting of estradiol, estriol, estradiol esters, progesterone and progestins to a patient in need thereof.

20. A method of making a compound according to any of claims 2-11, comprising reacting a ( 6 aR, 9 aS)-3-(phenylamino)-5-6a,7,8,9,9a-hexahydro-5-methyl 20 cyclopent[4,5]imidazo[l, 2 -ajpyrazolo[4,3-e]pyrimidin-4(2H)-one with a compound of formula Z-Y-X-L wherein L is a leaving group and X, Y and Z are as defined in claim 2, and isolating the compound according to any of claims 2-11 thus obtained.

21. The method of claim 20, wherein the ( 6 aR,9aS)-3-(phenylamino)-5-6a,7,8,9,9a 25 hexahydro-5-methyl-cyclopent[4,5]imidazo[1, 2 -a]pyrazolo[4,3-e]pyrimidin-4(2H) one is a compound of Formula QK: 4 0 'N'R NH N' N N 69 QK

22. A method of making a compound according to any of claims 1-11, comprising cyclizing a compound of Formula QV RI O HN__ NH HN N N Y_ 8 OH Z'x~ 5 with a dehydrating/halogenating agent.

23. The method according to claim 22, wherein said agent is SOCl 2 .

24. Use of the Compound according to any of claims 1-Il or a pharmaceutical 10 composition according to claim 12 for the manufacture of a medicament for the treatment or prophylactic treatment of the following diseases: Parkinson's disease, restless leg, tremors, dyskinesias, Huntington's disease, Alzheimer's disease, and drug-induced movement disorders; depression, attention deficit disorder, attention deficit hyperactivity disorder, bipolar illness, anxiety, sleep disorder, narcolepsy, 15 cognitive impairment, dementia, Tourette's syndrome, autism, fragile X syndrome, psychostimulant withdrawal, and/or drug addiction; cerebrovascular disease, stroke, congestive heart disease, hypertension, pulmonary hypertension, and/or sexual dysfunction; asthma, chronic obstructive pulmonary disease, and/or allergic rhinitis, as well as autoimmune and inflammatory diseases; and/or female sexual dysfunction, 20 exercise amenorrhoea, anovulation, menopause, menopausal symptoms, hypothyroidism, pre-menstrual syndrome, premature labor, infertility, irregular menstrual cycles, abnormal uterine bleeding, osteoporosis, multiple sclerosis, prostate enlargement, prostate cancer, hypothyroidism, estrogen-induced endometrial hyperplasia or carcinoma; and/or any disease or condition characterized by low levels 25 of cAMP and/or cGMP (or inhibition of cAMP and/or cGMP signaling pathways) in cells expressing PDE1, and/or by reduced dopamine DI receptor signaling activity; 70