CYCLOHEXAPEPTIDES AS SELECTIVE SOMATOSTATIN SST5 RECEPTOR AGONISTS
20210363188 · 2021-11-25
Inventors
- Giles Albert Brown (Cambridge, GB)
- Miles Stuart Congreve (Cambridge, GB)
- Conor Scully (Cambridge, GB)
- Rebecca Paul (Cambridge, GB)
- Andrea Bortolato (Cambridge, GB)
Cpc classification
A61P5/02
HUMAN NECESSITIES
C07K7/64
CHEMISTRY; METALLURGY
A61P35/00
HUMAN NECESSITIES
International classification
Abstract
The disclosures herein relate to novel compounds of formula (1): (1) and salts thereof, wherein W, X, Y, Z, m, n, q, R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5 and R.sup.6 are defined herein, and their use in treating, preventing, ameliorating, controlling or reducing the risk of disorders associated with somatostatin receptors.
##STR00001##
Claims
1. A compound of the formula (1): ##STR00114## or a salt thereof, wherein; W is CH or N; X and Y are CH.sub.2 or O, wherein one of X and Y is CH.sub.2 and the other of X and Y is O; Z is CHR.sup.7, NR.sup.8 or O; m is 1 or 2; n is 0 to 3; each R.sup.1 is independently selected from halo, C.sub.1-C.sub.3 alkyl and C.sub.1-C.sub.3 alkoxy, wherein the C.sub.1-C.sub.3 alkyl and alkoxy groups are optionally substituted with up to 6 fluorine atoms; q is 0 to 2; R.sup.2 is selected from H and C.sub.1-C.sub.3 alkyl optionally substituted with up to 6 fluorine atoms; R.sup.3 is selected from optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.3-C.sub.6 cycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; R.sup.4 is H or optionally substituted C.sub.1-C.sub.3 alkyl, where the C.sub.1-C.sub.3 alkyl group is optionally joined to R.sup.5 to form a ring; R.sup.5 is selected from optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted aryl and optionally substituted heteroaryl, where R.sup.5 is optionally joined to R.sup.4 to form a ring; R.sup.6 is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted O-aryl or optionally substituted O-heteroaryl; R.sup.7 is selected from H, optionally substituted C.sub.1-C.sub.6 alkyl, CONR.sup.10R.sup.11, OCONR.sup.10R.sup.11, OCOR.sup.10, OCOOR.sup.10, COOR.sup.10 or OR.sup.12; R.sup.8 is selected from H, CONR.sup.10R.sup.11 or COOR.sup.10; R.sup.10 and R.sup.11 are independently selected from H, optionally substituted C.sub.1-C.sub.6 alkyl and optionally substituted C.sub.2-C.sub.6 alkyl where any one atom in the C.sub.2-C.sub.6 alkyl group is replaced by a heteroatom selected from N, O and S, or wherein R.sup.10 and R.sup.11 are optionally joined to form a ring; and R.sup.12 is optionally substituted aryl or optionally substituted heteroaryl.
2. The compound according to claim 1 which is a compound of formula (1a): ##STR00115## or a salt thereof, wherein W, X, Y, Z, m, n, q, R.sup.1, R.sup.2, R.sub.3, R.sup.5 and R.sup.6 are as defined in claim 1.
3. The compound according to claim 1 which is a compound of formula (1b): ##STR00116## or a salt thereof, wherein W, Z, m, q, R.sup.1 and R.sup.3 are as defined in claim 1.
4. The compound according to any one of claims 1 to 3, wherein W is CH.
5. The compound according to any one of claims 1 to 3, wherein W is N.
6. The compound according to claim 1 or claim 2, wherein X is O and Y is CH.sub.2.
7. The compound according to any one of claims 1 to 6, wherein Z is CHR.sup.7.
8. The compound according to any one of claims 1 to 7, wherein R.sup.1 is OMe or Me.
9. The compound according to claim 1 or claim 2, wherein R.sup.2 is H.
10. The compound according to any one of claims 1 to 9, wherein R.sup.3 is optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.3-C.sub.6 cycloalkyl or optionally substituted aryl, wherein the optional substituents are selected from halo, C.sub.1-C.sub.3 alkyl and C.sub.1-C.sub.3 alkoxy, wherein the C.sub.1-C.sub.3 alkyl and alkoxy groups are themselves optionally substituted with up to 6 fluorine atoms.
11. The compound according to claim 10, wherein R.sup.3 is optionally substituted phenyl, optionally substituted cyclohexyl, optionally substituted cyclopentyl or optionally substituted cyclobutyl, wherein the optional substituents are selected from chloro, bromo and fluoro.
12. The compound according to claim 1 or claim 2, wherein R.sup.4 is H.
13. The compound according to claim 1 or claim 2, wherein R.sup.5 is optionally substituted aryl or optionally substituted heteroaryl, wherein the optional substituents are selected from halo, C.sub.1-C.sub.3 alkyl and C.sub.1-C.sub.3 alkoxy, wherein the C.sub.1-C.sub.3 alkyl and alkoxy groups are themselves optionally substituted with up to 6 fluorine atoms.
14. The compound according to claim 13, wherein R.sup.5 is optionally substituted phenyl or optionally substituted pyridyl, wherein the optional substituents are selected from chloro, bromo, fluoro and OMe.
15. The compound according to claim 1 or claim 2, wherein R.sup.4 and R.sup.5 are joined together to form a ring.
16. The compound according to claim 15, wherein the ring moiety formed by R.sup.4 and R.sup.5 is selected from the group consisting of: ##STR00117## wherein said ring moieties are optionally substituted with a group or groups selected from halo, C.sub.1-C.sub.3 alkyl and C.sub.1-C.sub.3 alkoxy, wherein the C.sub.1-C.sub.3 alkyl and alkoxy groups are themselves optionally substituted with up to 6 fluorine atoms.
17. The compound according to any one of claims 1 to 16, wherein R.sup.7 is OCONR.sup.10R.sup.11, COOR.sup.10 or OR.sup.12; wherein R.sup.10 and R.sup.11 are C.sub.2-C.sub.6 alkyl where any one atom in the C.sub.2-C.sub.6 alkyl group is replaced by a heteroatom selected from N, O and S, where R.sup.10 and R.sup.11 are optionally via CH.sub.2 to form a ring; and R.sup.12 is pyridyl.
18. The compound according to any one of claims 1 to 3, wherein the moiety formed by Z and m is selected from: ##STR00118##
19. The compound according to claim 1 or claim 2 wherein R.sup.6 is phenyl.
20. The compound according to claim 1 which is selected from the group consisting of: ##STR00119## ##STR00120## ##STR00121## ##STR00122## ##STR00123## ##STR00124## ##STR00125## ##STR00126## ##STR00127## ##STR00128## ##STR00129## ##STR00130## ##STR00131## ##STR00132## ##STR00133## ##STR00134## ##STR00135## ##STR00136## ##STR00137##
21. The compound according to claim 1 which is selected from the group consisting of: ##STR00138## ##STR00139##
22. The compound according to any one of claims 1 to 21 having SST.sub.5 receptor agonist activity.
23. The compound according to claim 22 which exhibits selectivity towards the SST.sub.5 receptor compared to the SST.sub.2 receptor.
24. A pharmaceutical composition comprising a compound as defined in any one of claims 1 to 23 and a pharmaceutically acceptable excipient.
25. The compound or composition according to any one of claims 1 to 24 for use in the treatment of Cushing's Disease, Cushing's Syndrome, Acromegaly, Neuroendocrine tumours (inc. Carcinoid tumours), Thyrotropinomas, Prolactinomas, Non-functioning pituitary adenomas, Nelson's syndrome, Congenital hyperinsulinism, Post-gastric bypass hypoglycaemia, Dumping syndrome, Hyperinsulinemic obesity, Insulinoma, Polycystic kidney disease, Polycystic liver disease, Portal hypertension, Ascites, Pancreatic cancer, Pancreatic fistula, Acute or chronic pancreatitis, Hepatocellular carcinoma, Irritable bowel syndrome/disease or Headache disorders (inc. migraine, cluster headache, tension-type headache).
Description
DETAILED DESCRIPTION OF THE INVENTION
[0045] The invention relates to novel compounds. The invention also relates to the use of novel compounds as agonists of the SST.sub.5 receptor. The invention further relates to the use of novel compounds in the manufacture of medicaments for use as SST.sub.5 receptor agonists or for the treatment of disorders associated with somatostatin receptors. The invention further relates to compounds, compositions and medicaments which are selective SST.sub.5 receptor agonists.
[0046] The invention further relates to compounds, compositions and medicaments useful for the treatment of Cushing's Disease, Cushing's Syndrome, Acromegaly, Neuroendocrine tumours (inc. Carcinoid tumours), Thyrotropinomas, Prolactinomas, Non-functioning pituitary adenomas, Nelson's syndrome, Congenital hyperinsulinism, Post-gastric bypass hypoglycaemia, Dumping syndrome, Hyperinsulinemic obesity, Insulinoma, Polycystic kidney disease, Polycystic liver disease, Portal hypertension, Ascites, Pancreatic cancer, Pancreatic fistula, Acute or chronic pancreatitis, Hepatocellular carcinoma, Irritable bowel syndrome/disease, Headache disorders (inc. migraine, cluster headache, tension-type headache), Nesidioblastosis, Neuropathic pain, Hyperalgesia, Causalgia, Acute pain, Burn pain, Atypical facial pain, Back pain, Complex regional pain syndrome I and II, Post-chemotherapy pain, Post-stroke pain, Post-operative pain, Type I diabetes mellitus, Type II diabetes mellitus, Diabetic retinopathy, Diabetic macular edema, Thyroid eye disease, Cystoid macular edema, Diabetic nephropathy, Diabetic neuropathy, Peptic ulcers, Enterocutaneous, Watery diarrhea syndrome, AIDS-related diarrhea, Chemotherapy-induced diarrhea and Gastrointestinal bleeding.
[0047] Compounds of the invention include compounds of the formula (1):
##STR00005##
[0048] or a salt thereof, wherein;
[0049] W is CH or N;
[0050] X and Y are CH.sub.2 or O, wherein one of X and Y is CH.sub.2 and the other of X and Y is O;
[0051] Z is CHR.sup.7, NR.sup.8 or O;
[0052] m is 1 or 2;
[0053] n is 0 to 3;
[0054] each R.sup.1 is independently selected from halo, C.sub.1-C.sub.3 alkyl and C.sub.1-C.sub.3 alkoxy, wherein the C.sub.1-C.sub.3 alkyl and alkoxy groups are optionally substituted with up to 6 fluorine atoms;
[0055] q is 0 to 2;
[0056] R.sup.2 is selected from H and C.sub.1-C.sub.3 alkyl optionally substituted with up to 6 fluorine atoms;
[0057] R.sup.3 is selected from optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.3-C.sub.6 cycloalkyl, optionally substituted aryl and optionally substituted heteroaryl;
[0058] R.sup.4 is H or optionally substituted C.sub.1-C.sub.3 alkyl, where the C.sub.1-C.sub.3 alkyl group is optionally joined to R.sup.5 to form a ring;
[0059] R.sup.5 is selected from optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted aryl and optionally substituted heteroaryl, where R.sup.5 is optionally joined to R.sup.4 to form a ring;
[0060] R.sup.6 is selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted O-aryl or optionally substituted O-heteroaryl;
[0061] R.sup.7 is selected from H, optionally substituted C.sub.1-C.sub.6 alkyl, CONR.sup.10R.sup.11, OCONR.sup.10R.sup.11, OCOR.sup.10, OCOOR.sup.10, COOR.sup.10 or OR.sup.12;
[0062] R.sup.8 is selected from H, CONR.sup.10R.sup.11 or COOR.sup.10;
[0063] R.sup.10 and R.sup.11 are independently selected from H, optionally substituted C.sub.1-C.sub.6 alkyl and optionally substituted C.sub.2-C.sub.6 alkyl where any one atom in the C.sub.2-C.sub.6 alkyl group is replaced by a heteroatom selected from N, O and S, or wherein R.sup.10 and R.sup.11 are optionally joined to form a ring; and
[0064] R.sup.12 is optionally substituted aryl or optionally substituted heteroaryl.
[0065] W can be CH or N. W can be CH. W can be N.
[0066] X and Y can be CH.sub.2 or O, wherein one of X and Y is CH.sub.2 and one of X and Y is O.
[0067] X can be CH.sub.2. X can be O.
[0068] Y can be CH.sub.2. Y can be O.
[0069] Z can be CHR.sup.7, NR.sup.8 or O. Z can be CHR.sup.7. Z can be NR.sup.8. Z can be O.
[0070] m can be 1 or 2. m can be 1. m can be 2.
[0071] n can be 0 to 3. n can be 0. n can be 1. n can be 2. n can be 3.
[0072] Each occurrence of R.sup.1 can be halo, C.sub.1-C.sub.3 alkyl or C.sub.1-C.sub.3 alkoxy, wherein the C.sub.1-C.sub.3 alkyl and alkoxy groups are optionally substituted with up to 6 fluorine atoms. R.sup.1 can be methoxy or methyl. R.sup.1 can be methoxy. R.sup.1 can be methyl. R.sup.1 can be OMe or Me. R.sup.1 can be OMe. R.sup.1 can be Me.
[0073] q can be 0 to 2. q can be 0. q can be 1. q can be 2.
[0074] R.sup.2 can be H or C.sub.1-C.sub.3 alkyl optionally substituted with up to 6 fluorine atoms. R.sup.2 can be hydrogen. R.sup.2 can be H. R.sup.2 can be methyl. R.sup.2 can be Me.
[0075] R.sup.3 can be optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.3-C.sub.6 cycloalkyl, optionally substituted aryl or optionally substituted heteroaryl. R.sup.3 can be optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.3-C.sub.6 cycloalkyl or optionally substituted aryl, wherein the optional substituents are selected from halo, C.sub.1-C.sub.3 alkyl and C.sub.1-C.sub.3 alkoxy, wherein the C.sub.1-C.sub.3 alkyl and alkoxy groups are themselves optionally substituted with up to 6 fluorine atoms. R.sup.3 can be optionally substituted phenyl, optionally substituted cyclohexyl, optionally substituted cyclopentyl or optionally substituted cyclobutyl, wherein the optional substituents are selected from chloro, bromo and fluoro. R.sup.3 can be phenyl, optionally substituted with chloro, bromo or fluoro. R.sup.3 can be phenyl.
[0076] R.sup.4 can be H or optionally substituted C.sub.1-C.sub.3 alkyl, where the C.sub.1-C.sub.3 alkyl group is optionally joined to R.sup.5 to form a ring. R.sup.4 can be hydrogen. R.sup.4 can be H. R.sup.4 can be optionally substituted C.sub.1-C.sub.3 alkyl. R.sup.4 can be C.sub.1-C.sub.3 alkyl. R.sup.4 can be joined to R.sup.5 to form a ring.
[0077] R.sup.5 can be optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted aryl or optionally substituted heteroaryl, where R.sup.5 is optionally joined to R.sup.4 to form a ring. R.sup.5 can be optionally substituted aryl or optionally substituted heteroaryl, wherein the optional substituents are selected from halo, C.sub.1-C.sub.3 alkyl and C.sub.1-C.sub.3 alkoxy, wherein the C.sub.1-C.sub.3 alkyl and alkoxy groups are themselves optionally substituted with up to 6 fluorine atoms. R.sup.5 can be optionally substituted phenyl or optionally substituted pyridyl, wherein the optional substituents are selected from chloro, bromo, fluoro and OMe. R.sup.5 can be phenyl. R.sup.5 can be pyridyl.
[0078] R.sup.5 can be joined to R.sup.4 to form a ring. R.sup.4 and R.sup.5 can be joined to form a ring selected from the group consisting of:
##STR00006##
[0079] wherein said ring moieties are optionally substituted with a group or groups selected from halo, C.sub.1-C.sub.3 alkyl and C.sub.1-C.sub.3 alkoxy, wherein the C.sub.1-C.sub.3 alkyl and alkoxy groups are themselves optionally substituted with up to 6 fluorine atoms.
[0080] R.sup.6 can be optionally substituted aryl, optionally substituted heteroaryl, optionally substituted O-aryl or optionally substituted O-heteroaryl. R.sup.6 can be phenyl.
[0081] R.sup.7 can be H, optionally substituted C.sub.1-C.sub.6 alkyl, CONR.sup.10R.sup.11, OCONR.sup.10R.sup.11, OCOR.sup.10, OCOOR.sup.10, COOR.sup.10 or OR.sup.12. R.sup.7 can be OCONR.sup.10R.sup.11, COOR.sup.10 or OR.sup.12; wherein R.sup.10 and R.sup.11 are C.sub.2-C.sub.6 alkyl where any one atom in the C.sub.2-C.sub.6 alkyl group is replaced by a heteroatom selected from N, O and S, where R.sup.10 and R.sup.11 are optionally via CH.sub.2 to form a ring.
[0082] R.sup.7 can be
##STR00007##
[0083] R.sup.7 can be
##STR00008##
[0084] R.sup.7 can be
##STR00009##
[0085] R.sup.8 can be H, CONR.sup.10R.sup.11 or COOR.sup.10. R.sup.8 can be hydrogen. R.sup.8 can be H.
[0086] R.sup.8 can be
##STR00010##
[0087] R.sup.10 can be H, optionally substituted C.sub.1-C.sub.6 alkyl or optionally substituted C.sub.2-C.sub.6 alkyl where any one atom in the C.sub.2-C.sub.6 alkyl group is replaced by a heteroatom selected from N, O and S. R.sup.10 can be hydrogen. R.sup.10 can be H. R.sup.10 can be —CH.sub.2CH.sub.2NH.sub.2.
[0088] R.sup.11 can be H, optionally substituted C.sub.1-C.sub.6 alkyl or optionally substituted C.sub.2-C.sub.6 alkyl where any one atom in the C.sub.2-C.sub.6 alkyl group is replaced by a heteroatom selected from N, O and S. R.sup.11 can be hydrogen. R.sup.11 can be H. R.sup.11 can be —CH.sub.2CH.sub.2NH.sub.2.
[0089] R.sup.10 and R.sup.11 can be joined to form a ring.
[0090] R.sup.12 can be optionally substituted aryl or optionally substituted heteroaryl. R.sup.12 can be pyridyl.
[0091] The moiety formed by Z and m can be selected from:
##STR00011##
[0092] The moiety formed by Z and m can be
##STR00012##
[0093] The moiety formed by Z and m can be
##STR00013##
[0094] Particular compounds of the invention also include compounds of formula (1a):
##STR00014##
[0095] or a salt thereof, wherein W, X, Y, Z, m, n, q, R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5 and R.sup.6 are as defined above.
[0096] Particular compounds of the invention also include compounds of formula (1b):
##STR00015##
[0097] or a salt thereof, wherein W, Z, m, q, R.sup.1 and R.sup.3 are as defined above.
[0098] The compound can be selected from any one of Examples 1 to 79 shown in Table 1.
[0099] Specific examples of compounds include compounds having somatostatin receptor agonist activity.
[0100] Specific examples of compounds include compounds having SST.sub.5 receptor agonist activity.
[0101] Specific examples of compounds include compounds that exhibit selectivity towards the SST.sub.5 receptor compared to the SST.sub.2 receptor.
[0102] The compounds of the invention may be used in a pharmaceutical composition comprising a compound of the invention and a pharmaceutically acceptable excipient.
[0103] The compounds of the invention may be used in medicine.
[0104] The compounds of the invention may be used in the treatment of disorders associated with somatostatin receptors.
[0105] The compounds of the invention may be used in the treatment of disorders associated with the SST.sub.5 receptor.
[0106] The compounds of the invention may be used in the treatment of Cushing's Disease, Cushing's Syndrome, Acromegaly, Neuroendocrine tumours (inc. Carcinoid tumours), Thyrotropinomas, Prolactinomas, Non-functioning pituitary adenomas, Nelson's syndrome, Congenital hyperinsulinism, Post-gastric bypass hypoglycaemia, Dumping syndrome, Hyperinsulinemic obesity, Insulinoma, Polycystic kidney disease, Polycystic liver disease, Portal hypertension, Ascites, Pancreatic cancer, Pancreatic fistula, Acute or chronic pancreatitis, Hepatocellular carcinoma, Irritable bowel syndrome/disease, Headache disorders (inc. migraine, cluster headache, tension-type headache).
Definitions
[0107] In this application, the following definitions apply, unless indicated otherwise.
[0108] The term “alkyl”, “aryl”, “halo”, “alkoxy”, “cycloalkyl” and “heteroaryl” are used in their conventional sense (e.g. as defined in the IUPAC Gold Book) unless indicated otherwise.
[0109] The term “treatment”, in relation to the uses of any of the compounds described herein, including those of the formula (1), formula (1a) or formula (1b), is used to describe any form of intervention where a compound is administered to a subject suffering from, or at risk of suffering from, or potentially at risk of suffering from the disease or disorder in question. Thus, the term “treatment” covers both preventative (prophylactic) treatment and treatment where measurable or detectable symptoms of the disease or disorder are being displayed.
[0110] The term “effective therapeutic amount” as used herein (for example in relation to methods of treatment of a disorder, disease or condition) refers to an amount of the compound which is effective to produce a desired therapeutic effect. For example, if the condition is pain, then the effective therapeutic amount is an amount sufficient to provide a desired level of pain relief. The desired level of pain relief may be, for example, complete removal of the pain or a reduction in the severity of the pain.
[0111] To the extent that any of the compounds described have chiral centres, the present invention extends to all optical isomers of such compounds, whether in the form of racemates or resolved enantiomers. The invention described herein relates to all crystal forms, solvates and hydrates of any of the disclosed compounds however so prepared. To the extent that any of the compounds disclosed herein have acid or basic centres such as carboxylates or amino groups, then all salt forms of said compounds are included herein. In the case of pharmaceutical uses, the salt should be seen as being a pharmaceutically acceptable salt.
[0112] Salts or pharmaceutically acceptable salts that may be mentioned include acid addition salts and base addition salts. Such salts may be formed by conventional means, for example by reaction of a free acid or a free base form of a compound with one or more equivalents of an appropriate acid or base, optionally in a solvent, or in a medium in which the salt is insoluble, followed by removal of said solvent, or said medium, using standard techniques (e.g. in vacuo, by freeze-drying or by filtration). Salts may also be prepared by exchanging a counter-ion of a compound in the form of a salt with another counter-ion, for example using a suitable ion exchange resin.
[0113] Examples of pharmaceutically acceptable salts include acid addition salts derived from mineral acids and organic acids, and salts derived from metals such as sodium, magnesium, potassium and calcium.
[0114] Examples of acid addition salts include acid addition salts formed with acetic, 2,2-dichloroacetic, adipic, alginic, aryl sulfonic acids (e.g. benzenesulfonic, naphthalene-2-sulfonic, naphthalene-1,5-disulfonic and p-toluenesulfonic), ascorbic (e.g. L-ascorbic), L-aspartic, benzoic, 4-acetamidobenzoic, butanoic, (+) camphoric, camphor-sulfonic, (+)-(1S)-camphor-10-sulfonic, capric, caproic, caprylic, cinnamic, citric, cyclamic, dodecylsulfuric, ethane-1,2-disulfonic, ethanesulfonic, 2-hydroxyethanesulfonic, formic, fumaric, galactaric, gentisic, glucoheptonic, gluconic (e.g. D-gluconic), glucuronic (e.g. D-glucuronic), glutamic (e.g. L-glutamic), α-oxoglutaric, glycolic, hippuric, hydrobromic, hydrochloric, hydriodic, isethionic, lactic (e.g. (+)-L-lactic and (±)-DL-lactic), lactobionic, maleic, malic (e.g. (−)-L-malic), malonic, (±)-DL-mandelic, metaphosphoric, methanesulfonic, 1-hydroxy-2-naphthoic, nicotinic, nitric, oleic, orotic, oxalic, palmitic, pamoic, phosphoric, propionic, L-pyroglutamic, salicylic, 4-amino-salicylic, sebacic, stearic, succinic, sulfuric, tannic, tartaric (e.g. (+)-L-tartaric), thiocyanic, undecylenic and valeric acids.
[0115] Also encompassed are any solvates of the compounds and their salts. Preferred solvates are solvates formed by the incorporation into the solid state structure (e.g. crystal structure) of the compounds of the invention of molecules of a non-toxic pharmaceutically acceptable solvent (referred to below as the solvating solvent). Examples of such solvents include water, alcohols (such as ethanol, isopropanol and butanol) and dimethylsulfoxide. Solvates can be prepared by recrystallising the compounds of the invention with a solvent or mixture of solvents containing the solvating solvent. Whether or not a solvate has been formed in any given instance can be determined by subjecting crystals of the compound to analysis using well known and standard techniques such as thermogravimetric analysis (TGA), differential scanning calorimetry (DSC) and X-ray crystallography.
[0116] The solvates can be stoichiometric or non-stoichiometric solvates. Particular solvates may be hydrates, and examples of hydrates include hemihydrates, monohydrates and dihydrates. For a more detailed discussion of solvates and the methods used to make and characterise them, see Bryn et al, Solid-State Chemistry of Drugs, Second Edition, published by SSCI, Inc of West Lafayette, Ind., USA, 1999, ISBN 0-967-06710-3.
[0117] The term “pharmaceutical composition” in the context of this invention means a composition comprising an active agent and comprising additionally one or more pharmaceutically acceptable carriers. The composition may further contain ingredients selected from, for example, diluents, adjuvants, excipients, vehicles, preserving agents, fillers, disintegrating agents, wetting agents, emulsifying agents, suspending agents, sweetening agents, flavouring agents, perfuming agents, antibacterial agents, antifungal agents, lubricating agents and dispersing agents, depending on the nature of the mode of administration and dosage forms. The compositions may take the form, for example, of tablets, dragees, powders, elixirs, syrups, liquid preparations including suspensions, sprays, inhalants, tablets, lozenges, emulsions, solutions, cachets, granules, capsules and suppositories, as well as liquid preparations for injections, including liposome preparations.
[0118] The compounds of the invention may contain one or more isotopic substitutions, and a reference to a particular element includes within its scope all isotopes of the element. For example, a reference to hydrogen includes within its scope .sup.1H, .sup.2H (D), and .sup.3H (T). Similarly, references to carbon and oxygen include within their scope respectively .sup.12C, .sup.13C and .sup.14C and .sup.16O and .sup.18O. In an analogous manner, a reference to a particular functional group also includes within its scope isotopic variations, unless the context indicates otherwise. For example, a reference to an alkyl group such as an ethyl group or an alkoxy group such as a methoxy group also covers variations in which one or more of the hydrogen atoms in the group is in the form of a deuterium or tritium isotope, e.g. as in an ethyl group in which all five hydrogen atoms are in the deuterium isotopic form (a perdeuteroethyl group) or a methoxy group in which all three hydrogen atoms are in the deuterium isotopic form (a trideuteromethoxy group). The isotopes may be radioactive or non-radioactive.
[0119] Therapeutic dosages may be varied depending upon the requirements of the patient, the severity of the condition being treated, and the compound being employed. Determination of the proper dosage for a particular situation is within the skill of the art. Generally, treatment is initiated with the smaller dosages which are less than the optimum dose of the compound. Thereafter the dosage is increased by small increments until the optimum effect under the circumstances is reached. For convenience, the total daily dosage may be divided and administered in portions during the day if desired.
[0120] The magnitude of an effective dose of a compound will, of course, vary with the nature of the severity of the condition to be treated and with the particular compound and its route of administration. The selection of appropriate dosages is within the ability of one of ordinary skill in this art, without undue burden. In general, the daily dose range may be from about 10 μg to about 30 mg per kg body weight of a human and non-human animal, preferably from about 50 μg to about 30 mg per kg of body weight of a human and non-human animal, for example from about 50 μg to about 10 mg per kg of body weight of a human and non-human animal, for example from about 100 μg to about 30 mg per kg of body weight of a human and non-human animal, for example from about 100 μg to about 10 mg per kg of body weight of a human and non-human animal and most preferably from about 100 μg to about 1 mg per kg of body weight of a human and non-human animal.
Pharmaceutical Formulations
[0121] While it is possible for the active compound to be administered alone, it is preferable to present it as a pharmaceutical composition (e.g. formulation).
[0122] Accordingly, in another embodiment of the invention, there is provided a pharmaceutical composition comprising at least one compound of the formula (1) as defined above together with at least one pharmaceutically acceptable excipient.
[0123] The composition may be a composition suitable for injection. The injection may be intra-venous (IV) or subcutaneous. The composition may be supplied in a sterile buffer solution or as a solid which can be suspended or dissolved in sterile buffer for injection.
[0124] The pharmaceutically acceptable excipient(s) can be selected from, for example, carriers (e.g. a solid, liquid or semi-solid carrier), adjuvants, diluents (e.g solid diluents such as fillers or bulking agents; and liquid diluents such as solvents and co-solvents), granulating agents, binders, flow aids, coating agents, release-controlling agents (e.g. release retarding or delaying polymers or waxes), binding agents, disintegrants, buffering agents, lubricants, preservatives, anti-fungal and antibacterial agents, antioxidants, buffering agents, tonicity-adjusting agents, thickening agents, flavouring agents, sweeteners, pigments, plasticizers, taste masking agents, stabilisers or any other excipients conventionally used in pharmaceutical compositions.
[0125] The term “pharmaceutically acceptable” as used herein means compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of a subject (e.g. a human subject) without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. Each excipient must also be “acceptable” in the sense of being compatible with the other ingredients of the formulation.
[0126] Pharmaceutical compositions containing compounds of the formula (1) can be formulated in accordance with known techniques, see for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., USA.
[0127] Suitable formulations typically contain 0-20% (w/w) buffers, 0-50% (w/w) cosolvents, and/or 0-99% (w/w) Water for Injection (WFI) (depending on dose and if freeze dried). Formulations for intramuscular depots may also contain 0-99% (w/w) oils.
[0128] The compounds of the formula (1) will generally be presented in unit dosage form and, as such, will typically contain sufficient compound to provide a desired level of biological activity. For example, a formulation may contain from 1 nanogram to 2 grams of active ingredient, e.g. from 1 nanogram to 2 milligrams of active ingredient. Within these ranges, particular sub-ranges of compound are 0.1 milligrams to 2 grams of active ingredient (more usually from 10 milligrams to 1 gram, e.g. 50 milligrams to 500 milligrams), or 1 microgram to 20 milligrams (for example 1 microgram to 10 milligrams, e.g. 0.1 milligrams to 2 milligrams of active ingredient).
[0129] The active compound will be administered to a patient in need thereof (for example a human or animal patient) in an amount sufficient to achieve the desired therapeutic effect (effective amount). The precise amounts of compound administered may be determined by a supervising physician in accordance with standard procedures.
EXAMPLES
[0130] The invention will now be illustrated, but not limited, by reference to the specific embodiments described in the following examples.
Examples 1 to 79
[0131] The compounds of Examples 1 to 79 shown in Table 1 below have been prepared. Their NMR and LCMS properties and the methods used to prepare them are set out in Table 2 and 3. The starting materials for each of the Examples are commercial unless indicated.
TABLE-US-00001 TABLE 1
General Procedures
[0132] Where no preparative routes are included, the relevant intermediate is commercially available. Commercial reagents were utilized without further purification. Room temperature (rt) refers to approximately 20-27° C. .sup.1H NMR spectra were recorded at 600 MHz on a Bruker instrument. Chemical shift values are expressed in parts per million (ppm), i.e. (δ)-values. The following abbreviations are used for the multiplicity of the NMR signals: s=singlet, br=broad, d=doublet, t=triplet, q=quartet, quint=quintet, td=triplet of doublets, tt=triplet of triplets, qd=quartet of doublets, ddd=doublet of doublet of doublets, ddt=doublet of doublet of triplets, m=multiplet. Coupling constants are listed as J values, measured in Hz. NMR and mass spectroscopy results were corrected to account for background peaks. Chromatography refers to column chromatography performed using 60-120 mesh silica gel and executed under nitrogen pressure (flash chromatography) conditions.
Analytical Methods
[0133] LCMS analysis of compounds was performed under electrospray conditions.
LCMS Method A
[0134] Water Acquity UPLC-H Class, Column: Waters BEH C18 1.7 μm, 2.1×50 mm. Gradient [time (min)/solvent B (%)]: 0.0/10, 0.5/10, 2.5/90, 3.0/90. (Solvent A—0.025% HCOOH in water; Solvent B—0.025% HCOOH in MeCN); Injection volume 0.1 μL (may vary); UV detection 210 nm; Column temperature 30° C.; 0.3 mL/min.
Analytical Method B
[0135] MS ion determined using LCMS method below under electrospray conditions, HPLC retention time (R.sub.T) determined using HPLC method below, purity >95% by HPLC unless indicated.
[0136] LCMS: Water Acquity UPLC-H Class, Column: Waters BEH C18 1.7 μm, 2.1×50 mm. Gradient [time (min)/solvent B (%)]: 0.0/10, 0.5/10, 2.5/90, 3.0/90. (Solvent A—0.025% HCOOH in water; Solvent B—0.025% HCOOH in MeCN); Injection volume 0.1 μL (may vary); UV detection 210 nm; Column temperature 30° C.; 0.3 mL/min.
[0137] HPLC: Agilent Technologies 1200, Water SunFire C18, 3.5 μm, 4.6×150 mm, 30° C., 1 mL/min; mobile phase A: 0.025% TFA in water, mobile phase B: MeCN. a: Gradient, 5-75% (MeCN:H2O, 15 min), b: Gradient, 10-90% (MeCN:H2O, 15 min); c: 20-90% (MeCN:H2O, 15 min).
Analytical Method C
[0138] MS ion determined using LCMS method below under electrospray conditions, HPLC retention time (R.sub.T) determined using HPLC method below, purity >95% by HPLC unless indicated.
[0139] LCMS: Agilent 1200 HPLC&6410B Triple Quad, Column: Xbridge C18 3.5 um 2.1*30 mm. Gradient [time (min)/solvent B (%)]: 0.0/10, 0.9/80, 1.5/90, 8.5/5, 1.51/10. (Solvent A=1 mL of TFA in 1000 mL Water; Solvent B=1 mL of TFA in 1000 mL of MeCN); Injection volume 5 μL (may vary); UV detection 220 nm 254 nm 210 nm; Column temperature 25° C.; 1.0 mL/min.
[0140] HPLC: Agilent Technologies 1200, Column: Gemini-NX C18 5 um 110 A 150*4.6 mm. Gradient [time (min)/solvent B (%)]: 0.0/30, 20/60, 20.1/90, 23/90. (Solvent A=1 mL of TFA in 1000 mL Water; Solvent B=1 mL of TFA in 1000 mL of MeCN); Injection volume 5 μL (may vary); UV detection 220 nm 254 nm; Column temperature 25° C.; 1.0 mL/min
Analytical Method D
[0141] Instrument: Thermo Scientific Orbitrap Fusion; Column: Phenomenex Kinetex Biphenyl 100 Å, 2.6 μm, 2.1×50 mm; Gradient [time (min)/solvent B in A (%)]: 0.00/10, 0.30/10, 0.40/60, 1.10/90, 1.70/90, 1.75/10, 1.99/10, 2.00/10; Solvents: Solvent A=0.1% formic acid in water; Solvent B=0.1% formic acid in acetonitrile; Injection volume 5 μL; Column temperature 25° C.; Flow rate 0.8 mL/min.
Synthesis of Intermediates
Synthesis of Fmoc-D-Trp(4-Me)
[0142] ##STR00095##
Procedure
N-Boc-4-methylindole (1)
[0143] 4-Methylindole (5.0 g, 38 mmol) and dimethylaminopyridine (1.04 g, 8.54 mmol) were dissolved in acetonitrile (60 mL) and then a solution of di-tert-butyl dicarbonate (14.0 g, 64 mmol) in acetonitrile (10 mL) was added slowly. The reaction was stirred for 18 h at room temperature under a N.sub.2 atmosphere then concentrated. The residue was diluted with EtOAc (100 mL), washed with 1N HCl (100 mL×3), saturated NaHCO.sub.3 (100 mL×3), water (100 mL×1) and brine (100 mL×1) and then dried (MgSO4). After filtration, the solvents were removed to afford compound 1 (8.3 g, quantitative yield), which was used for next step without further purification.
N-Boc-3-bromo-4-methylindole (2)
[0144] N-Bromosuccinimide (8.3 g, 47 mmol) was added all at once to a solution of N-Boc-indole (8.3 g, 38 mmol) in THF (100 mL) and the reaction was stirred for 18 h at room temperature. The reaction mixture was concentrated and the residue was diluted with EtOAc (100 mL), washed with saturated aqueous sodium metabisulfite (100 mL×3), saturated NaHCO.sub.3 aq (100 mL×3) and brine. The organic phase was dried (MgSO4) and concentrated to afford compound 2 (8.7 g, yield 74%) as an yellow oil.
(S)-Methyl 2-(tert-butoxycarbonylamino)-3-iodopropanoate (3)
[0145] A mixture of triphenylphosphine (131 g, 0.50 mol) and imidazole (34 g, 0.50 mol) in DCM (600 mL) was cooled to 0° C. and iodide (127 g, 0.50 mol) was added in small portions over 0.5 h. The cooling bath was removed and the mixture was stirred for 0.5 h. After the mixture was re-cooled to 0° C., a solution of compound 2 (73 g, 0.33 mol) in DCM (300 mL) was added dropwise. Upon completion, the cooling bath was removed and the mixture was allowed to warm to room temperature and stirred for 1.5 h. The mixture was filtered and the filtrate was concentrated to remove most of the solvents. MTBE (400 mL) was added to the residue and the mixture was filtered to remove triphenylphosphine oxide. The filtrate was concentrated and the residue was purified by flash column chromatography to afford 3 (74.0 g, 68% yield) as a yellow solid.
(R)-tert-Butyl 3-(2-(tert-butoxycarbonylamino)-3-methoxy-3-oxopropyl)-4-methyl-1H-indole-1-carboxylate (4)
[0146] A solution of compound 3 (7.5 g, 26 mmol) and iodine (0.5 g) in DMF (30 mL) was added to a suspension of zinc (4.5 g, 77 mmol) in DMF (50 mL). The mixture was stirred at 30° C. under nitrogen for 30 min, then cooled to room temperature. Compound 2 (8.0 g, 25.8 mol), S-Phos (200 mg) and Pd(dba).sub.2 (400 mgl) was added. The reaction mixture was stirred at 50° C. under nitrogen overnight and then cooled to room temperature. Brine (500 mL) was added and the resulting mixture was extracted with EtOAc (300 mL×3). The organics were combined, washed with brine and concentrated. The residue was purified by flash column chromatography (petroleum ether/ethyl acetate=100:1 to 40:1) to afford 4 as a viscous oil (3.3 g, 30% yield).
D-Trp(4-Me)-OH (5)
[0147] To a solution of compound 4 (3.3 g, 7.6 mmol) in a mixture of water/methanol (30 mL, 2:1) was added lithium hydroxide hydrate (1.3 g, 30 mmol). The reaction mixture was stirred at room temperature for 3 h and then concentrated to remove most of methanol. The residue was extracted with EtOAc (30 mL×3), washed with brine (20 mL×2), dried and concentrated to afford acid.
[0148] The above acid was dissolved in DCM (20 mL), and TFA (5 mL) was added. The reaction mixture was stirred for 1 h and then concentrated to afford 5 as a TFA salt, which was used in the next step without further purification.
Fmoc-D-Trp(4-Me)
[0149] Compound 5 (7.6 mmol) was dissolved in a mixture of acetone (100 mL) and saturated NaHCO.sub.3 aq (100 mL). Fmoc-OSu (2.5 g, 7.5 mmol) was added. The reaction mixture was stirred 18 h at room temperature. The reaction was diluted with H.sub.2O (100 mL), washed with hexane (100 mL×2). The aqueous phase was acidified with 1N HCl to pH 3 and extracted with EtOAc (100 mL×3). The combined EtOAc phases were washed with 1N HCl (100 mL×3) and brine (100 mL×1) and then dried over MgSO4. After filtration, the solvents was removed by concentration, the reside was purified by flash column chromatography (DCM/MeOH=100:1 to 20:1) to afford Fmoc-D-Trp(4-Me) (2.3 g, 68% yield form compound 4) as a white solid.
[0150] LCMS (Method A): m/z 441.7 [M+H].sup.+ (ES+)
[0151] The following compounds were synthesized using the same method:
TABLE-US-00002 Compound Structure Molecular Formula ESI Fmoc-D-Trp(5-Me)
Synthesis of Fmoc-Tyr(O-cyclohexylmethyl)
[0152] ##STR00099##
Procedure
Fmoc-L-Tyr(O-cyclohexylmethyl)-OMe (1)
[0153] A mixture of Fmoc-Tyr-OMe (1.5 g, 3.6 mmol), cyclohexylmethanol (500 mg, 4.4 mmol) and triphenylphosphine (1.13 g, 4.3 mol) in THF (100 mL) was cooled to 0° C. and DEAD (800 mg, 4.3 mol) was added. The mixture was allowed to warm to room temperature and stirred overnight. The reaction mixture was concentrated and the residue was purified by flash column chromatography to afford compound 1 (0.9 g, 48% yield).
Fmoc-L-Tyr(O-cyclohexylmethyl)
[0154] To a solution of compound 1 (0.9 g, 1.7 mmol) in water/methanol (30 mL, 10:1) was added lithium hydroxide hydrate (100 mg, 2.4 mmol). The reaction mixture was stirred at 0° C. for 3 h and then acidified with 1N HCl to pH 3. Then the mixture was extracted with EtOAc (30 mL×3), the extracts were washed with brine (20 mL×2), dried and concentrated. The reside was purified by flash column chromatography (DCM/MeOH=100:1 to 20:1) to afford Fmoc-L-Tyr(O-cyclohexylmethyl) (630 mg, 74% yield) as a white solid.
[0155] LCMS (Method A): m/z 500.9 [M+H].sup.+ (ES+),
[0156] The following compound was synthesized using the same method:
TABLE-US-00003 Compound Structure Molecular Formula ESI Fmoc-L-Tyr(O-4- chlorobenzyl)
Synthesis of Cyclohexapeptide Examples 1-79
[0157] Standard Fmoc solid phase peptide synthesis (SPPS) was used to synthesize the linear peptides which were then cleaved from the resin and cyclized to give the cyclohexapeptide. This linear synthesis can start at any position in the 6-mer sequence and then cyclisation can be carried out to give the final cyclohexapeptides, two methods are outlined below.
Method a
Peptide Synthesis
[0158] 1) To the vessel containing CTC-resin (0.4 mmol) and Fmoc-trans-4-HYP-OH (282.7 mg, 0.8 mmol, 2.0 eq.) under bubbled N.sub.2 (g) was added DCM (10 mL). [0159] 2) DIEA (263.9 μL, 1.6 mmol, 4.0 eq.) was added dropwise and the reaction stirred for 2 h. [0160] 3) MeOH (0.8 mL) was added and the reaction stirred for 30 min. [0161] 4) The resin was drained and washed with DMF (×5). [0162] 5) Bis(p-nitrophenyl) carbonate (730.0 mg, 2.4 mmol, 6.0 eq.) and DIEA (0.8 mL, 4.8 mmol, 12.0 eq.) were added to the vessel, and the reaction continued for 4 h. After the reaction was complete, the resin was washed with DMF (×5). [0163] 6) N-Boc-ethylenediamine (770.0 mg, 4.8 mmol, 12.0 eq.) and DIEA (0.8 mL, 4.8 mmol, 12.0 eq.) were added respectively to the resin in DMF (10 mL). The reaction was mixed overnight at room temperature. [0164] 7) The resin was drained and washed with DMF (×5). [0165] 8) A solution of piperidine in DMF (piperidine: DMF, 1:4) was added and the reaction continued for 30 min. [0166] 9) The resin was drained and washed with DMF (×5). [0167] 10) Fmoc-amino acid solution (0.8 mmol, 2.0 eq.) was added to the resin and mixed for 30 seconds, followed by adding HATU (2.0 eq.) and DIEA (4.0 eq.). N.sub.2 (g) was bubbled through the mixture for 1 h. [0168] 11) The resin was drained and washed with DMF (×5). [0169] 12) Repeat steps 8-11 for next amino acid coupling.
Peptide Cleavage, Cyclization and Purification
[0170] 1) To the flask containing the side chain protected peptide at rt was added cleavage buffer (1% TFA in DCM, 10 mL) and the resulting mixture stirred for 0.5 h. [0171] 2) The reaction mixture was filtered and the filtrate diluted with anhydrous DCM to 1 mM. DIEA was added to adjust the pH to 8. To the solution was added TBTU (385.2 mg, 1.2 mmol, 3.0 eq.) and HOBT (162.9 mg, 1.2 mmol, 3.0 eq.) and the resulting solution reacted for 3-4 h. [0172] 3) Upon completion, the reaction mixture was washed with 1 N HCl (200 mL) and the organic phase dried under vacuum to obtain the crude peptide, which was further treated with TFA:H.sub.2O:TIPS, 95:2.5:2.5 for 10 min-1 h. [0173] 4) The crude peptide was precipitated out by addition of methyl tert-butyl ether. [0174] 5) The crude peptide was purified by HPLC to give the final product. Prep-HPLC Conditions: Instrument: Gilson 281. Solvent: A—0.1% TFA in H2O, B—acetonitrile, Column: Luna C18 (200×25 mm; 10 μm) and Gemini C18 (150*30 mm; 5 μm) in series. Gradient [time (min)/solvent B (%)]: 0.0/25, 60.0/55, 60.1/90, 70/90, 70.1/10 for example 17.
Method b
Peptide Synthesis
[0175] 1) The CTC-resin (1 eq.) was swelled in DMF (10×) for 2 h, washed with DMF (10×) three times, DCM (10×) three times and DMF (10×) three times. [0176] 2) Fmoc-protected amino acid (3 eq.) was dissolved in DMF, and then DIC (2.9 eq.), HOBt (2.9 eq.), and NMM (5.6 eq.) was added. The resulting mixture was shaken for 1 min and transferred to the resin. [0177] 3) The reaction was shaken for 1-2 h and monitored by Kaiser method. [0178] 4) After coupling, the resin was washed with DMF. [0179] 5) The Fmoc group was removed using a solution of piperidine in DMF (piperidine: DMF, 1:4), twice (2 min, 8 min). [0180] 6) The resins were carefully washed with DMF (10×) three times, DCM (10×) three times and DMF (10×) three times.
[0181] Note: Fmoc-phenylglycine was transformed into its activated HOBt-ester [fresh prepared from Fmoc-Phg (3.0 eq.) and HOBt (3.0 eq.) in the presence of DIC (3.0 eq., 0° C., 20 min) before solid phase synthesis.
Peptide Cleavage, Cyclization and Purification
[0182] 1) The full-length, linear peptides were cleaved from the resin by swelling and shaking the peptide-resin for 4 h in a 1:1 (v:v) TFE/DCM (10 vol/g of dried resin). [0183] 2) The cleaved solution was filtered and the drained resin was washed with additional DCM (5 vol/g of initial dried peptide-resin) to fully extract the cleaved peptide from the resin. Solvents in the combined filtrates were evaporated and the residue dried. [0184] 3) The crude peptides were dissolved in a mixture of DMF/DCM (30 vol/g, ⅕), cooled to 0° C. and DIC (5 eq.) and HOBt (5 eq.) were added. Upon completion, the reaction was washed with aq. sat. NaCO.sub.3 (2×) and brine (1×). The organic layer was dried, filtered, and concentrated to afford crude cyclopeptides. [0185] 4) Crude cyclopeptides were deprotected by stirring in a mixture of TFA/DCM (50 vol/g, 1/1) for 1 h. The products were precipitated from cold diethyl ether and filtered to afford crude peptides, which were purified by prep-HPLC to afford cyclic hexapeptides. Prep-HPLC Conditions: Instrument: Water Auto-P 2545/2489/515. Solvent: A—0.1% TFA in Water, B—MeCN, Column: SunFire Prep C18 OBD, 5 μm, 30×100 mm. Gradient [time (min)/solvent B (%)]: 0.0/30, 1.0/30, 9.0/45, 9.5/95, 13.0/95. UV detection 210 nm & MS detection; Column temperature 25° C.; 30 mL/min.
TABLE-US-00004 TABLE 2 HRMS and LCMS properties and the methods used to prepare and purify peptides represented by Examples 1-79 Synthetic Analytical Example Method HRMS (Method D) Method LCMS/HPLC 1 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1073.5 [M + H].sup.+, C.sub.60H.sub.70O.sub.9N.sub.10 537.2658; Found 537.2672 R.sub.T = 6.35 min (b) 2 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1087.9 [M + H].sup.+, C.sub.61H.sub.72O.sub.9N.sub.10 544.2736; Found 544.2750 R.sub.T = 8.32 min (b) 3 a (HESI/FT) m/z: [M + 2H].sup.3+ Calcd for C m/z 1103.3 [M + H].sup.+, C.sub.61H.sub.74O.sub.9N.sub.11 368.18848; Found 368.1884; R.sub.T = 11.07 min 3 a m/z: [M + 2H].sup.2+ Calcd for C.sub.61H.sub.73O.sub.9N.sub.11 551.77909; Found 551.7786; m/z: [M + H] Calcd for C.sub.6H.sub.72O.sub.9N.sub.11 1102.5509; Found 1102.5507 4 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1087.9 [M + H].sup.+, C.sub.61H.sub.72O.sub.9N.sub.10 544.2736; Found 544.2750 R.sub.T = 8.10 min (b) 5 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1103.78 [M + H].sup.+, C.sub.61H.sub.72O.sub.10N.sub.10 552.2711; Found 552.2724 R.sub.T = 7.85 min (b) 6 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1108.82 [M + H].sup.+, C.sub.60H.sub.69O.sub.9N.sub.10Cl 554.2463; Found 554.2477 R.sub.T = 7.56 min (b) 7 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1108.82 [M + 2H].sup.+, C.sub.60H.sub.69O.sub.9N.sub.10Cl 554.2463; Found 554.2477 R.sub.T = 7.62 min (b) 8 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 554.9 [M + 2H].sup.2+, C.sub.60H.sub.69O.sub.9N.sub.10Cl 554.2463; Found 554.2477 R.sub.T = 7.76 min (b) 9 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1104.14 [M + H].sup.+, C.sub.61H.sub.72O.sub.10N.sub.10 552.2711; Found 552.2724 R.sub.T = 8.87 min (a) 10 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1152.74 [M + H].sup.+, C.sub.60H.sub.69O.sub.9N.sub.10Br 576.2211; Found 576.2225 R.sub.T = 9.49 min (a) 11 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1096.8 [M + Na].sup.+, C.sub.59H.sub.69O.sub.9N.sub.11 537.7634; Found 537.7634 R.sub.T = 6.62 min (b) 12 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1103.5 [M + H].sup.+, C.sub.61H.sub.72O.sub.10N.sub.10 552.2711; Found 552.2711 R.sub.T = 7.54 min (b) 13 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1087.8 [M + H].sup.+, C.sub.61H.sub.72O.sub.9N.sub.10 544.2736; Found 544.2750 R.sub.T = 8.11 min (b) 14 a (HESI/FT) m/z: [M + H].sup.2+ Calcd for C m/z 552.7 [M + 2H].sup.2+, C.sub.60H.sub.73O.sub.9N.sub.12 368.52023; Found 368.5201; R.sub.T = 7.08 min m/z: [M + H].sup.2+ Calcd for C.sub.60H.sub.72O.sub.9N.sub.12 552.27671; Found 552.2763; m/z: [M + H].sup.+ Calcd for C.sub.60H.sub.71O.sub.9N.sub.12 1103.54615; Found 1103.5452 15 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for C m/z 1102.2 [M + H].sup.+, C.sub.6H.sub.73O.sub.9N.sub.11 551.7791; Found 551.7791 R.sub.T = 9.83 min 16 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for C m/z 1103.4 [M + H].sup.+, C.sub.60H.sub.72O.sub.9N.sub.12 552.2767; Found 552.2781 R.sub.T = 9.74 min 17 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for C m/z 1113.5 [M + H].sup.+, C.sub.63H.sub.74O.sub.9N.sub.10 557.28146; Found 557.2812; R.sub.T = 10.89 min m/z: [M + H].sup.+ Calcd for C.sub.63H.sub.73O.sub.9N.sub.10 1113.55565; Found 1113.5544 18 a (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for C m/z 557.3 + [M + 2H].sup.2+, C.sub.8H.sub.74O.sub.9N.sub.18 557.2815; Found 557.2828 R.sub.T = 10.67 min 19 a (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for C m/z 557.3 [M + 2H].sup.2+, C.sub.83H.sub.74O.sub.9N.sub.10 557.2815; Found 557.2828 R.sub.T = 10.37 min 20 a (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for C m/z 1093.4 [M + H].sup.+, C.sub.83H.sub.70O.sub.8N.sub.18 547.2684; Found 547.2684 R.sub.T = 13.42 min 21 a (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for C m/z 1113.3 [M + H].sup.+, C.sub.63H.sub.72O.sub.9N.sub.10Na 1135.54; Found 11135.5378; R.sub.T = 9.60 min m/z: [M + H].sup.+ Calcd for C.sub.63H.sub.73O.sub.9N.sub.10 1113.5557; Found 1113.5556 22 a (HESI/FT) m/z: [M + 2H].sup.+ Calcd for C m/z 1093.4 [M + H].sup.+, C.sub.83H.sub.70O.sub.8N.sub.10 547.2684; Found 547.2684 R.sub.T = 10.37 min 23 a (HESI/FT) m/z: [M + 2H].sup.+ Calcd for C m/z 1093.4 [M + H].sup.+, C.sub.83H.sub.70O.sub.8N.sub.10 547.2684; Found 547.2684 R.sub.T = 10.27 min 24 a (HESI/FT) m/z: [M + H].sup.+ Calcd for C m/z 1093.3 [M + H].sup.+, C.sub.83H.sub.70O.sub.8N.sub.10 547.2684; Found 547.2697 R.sub.T = 10.65 min 25 a (HESI/FT) m/z: [M + H].sup.+ Calcd for C m/z 1114.4 [M + H].sup.+ , C.sub.62H.sub.72O.sub.9N.sub.111114.5509; Found R.sub.T = 9.56 min 1136.5330 m/z: [M + H].sup.+ Calcd for C.sub.62H.sub.72O.sub.9N.sub.11 1114.5509; Found 1114.5508 26 b (HESI/FT) m/z: [M + H].sup.2+ Calcd for C m/z 1114.3 [M + H].sup.+ , C.sub.62H.sub.73O.sub.9N.sub.11 557.77909; Found 557.7789; R.sub.T = 9.42 min m/z: [M + H].sup.+ Calcd for C.sub.62H.sub.72O.sub.9N.sub.11 1114.5509; Found 1114.5502 27 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1102.8 [M + Na].sup.+ , C.sub.59H.sub.75O.sub.9N.sub.11 540.7869; Found 540.7883 R.sub.T = 8.45 min (b) 28 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1102.8 [M + H].sup.+ , C.sub.62H.sub.74O.sub.9N.sub.10 551.2815; Found 551.2828 R.sub.T = 8.25 min (b) 29 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1102.0 [M + H].sup.+, C.sub.62H.sub.74O.sub.9N.sub.10 551.2815; Found 551.2815 R.sub.T = 8.55 min (b) 30 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1136.8 [M + H].sup.+, C.sub.61H.sub.72O.sub.9N.sub.11Cl 568.7596; Found 568.7609 R.sub.T = 8.80 min (b) 31 b (HESI/FT) m/z: [M + H].sup.2+ Calcd for B m/z 1028.9 [M + H].sup.+, C.sub.59H.sub.66O.sub.8N.sub.9 1028.50289; Found R.sub.T = 10.72 min (b) 1028.5022; m/z: [M + H].sup.2+ Calcd for C.sub.59H.sub.67O.sub.8N.sub.9 514.75508; Found 514.7549 32 b (HESI/FT) m/z:[M + H].sup.+ Calcd for B m/z 1041.8 [M + H].sup.+ , C.sub.61H.sub.69O.sub.8N.sub.8 1041.5233; Found 1041.5233 R.sub.T = 11.26 min (b) 33 a (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for m/z 1103.3 [M + H].sup.+ , C881-17209N12 552.2767; Found 552.2767 C R.sub.T = 9.17 min 34 a (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for C m/z 1103.3 [M + H].sup.+ , C881-17209N12 552.2767; Found 552.2767 R.sub.T = 9.91 min 35 a (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for C m/z 1114.4 [M + H].sup.+ , C.sub.62H.sub.73O.sub.9N.sub.11 557.7791; Found 557.7791 R.sub.T = 10.93 min 36 a (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for C m/z 1114.4 [M + H].sup.+ , C821-17309N11 557.7791; Found 557.7804 R.sub.T = 10.00 min 37 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1052.1 [M + H].sup.+ , C.sub.57H.sub.71O.sub.9N.sub.11 526.764; Found 526.2757 R.sub.T = 7.44 min (b) 38 b (HESI/FT) m/z:[M + 2H].sup.2+ Calcd for B m/z 1101.9 [M + H].sup.+, C.sub.62H.sub.74O.sub.9N.sub.10 551.2815; Found 551.2815 R.sub.T = 7.04 min (c) 39 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1155.0 [M + Na].sup.+ , C.sub.62H.sub.75O.sub.10N.sub.11 566.7844; Found 566.7857 R.sub.T = 6.23 min (c) 40 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1159.0 [M + Na].sup.+ , C.sub.61H.sub.72O.sub.9N.sub.11Cl 568.7596; Found 568.7609 R.sub.T = 7.45 min (b) 41 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1136.0 [M + H].sup.+ , C.sub.62H.sub.73O.sub.9N.sub.10Cl 568.2620; Found 568.2620 R.sub.T = 8.09 min (b) 42 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1157.9 [M + Na].sup.+, C.sub.62H.sub.73O.sub.9N.sub.10Cl 568.2612; Found 568.2634 R.sub.T = 8.17 min (b) 43 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1131.9 [M + H].sup.+, C.sub.63H.sub.76O.sub.10N.sub.10 566.2868; Found 566.2881 R.sub.T = 6.92 min (c) 44 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1120.0 [M + H].sup.+, C.sub.63H.sub.80O.sub.9N.sub.10 560.30494; Found 560.3056 R.sub.T = 7.57 min (c) 45 b (HESI/FT) m/z: [M + H].sup.+ Calcd for B m/z 1028.2 [M + H].sup.+, C.sub.60H.sub.67O.sub.8N.sub.8 1027.5076; Found 1027.5078 m/z: [3M + H].sup.2+ Found 1541 46 b (HESI/FT) m/z: [M + H].sup.+ Calcd for B m/z 1058.0 [M + H].sup.+, C.sub.61H.sub.69O.sub.9N.sub.8 1057.5182; Found 1057.5239 R.sub.T = 10.38 min (b) 47 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1171.9 [M + H].sup.+, C.sub.61H.sub.71O.sub.9N.sub.11Cl.sub.2 585.7401; Found 585.7408 R.sub.T = 8.12 min (b) 48 (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1158.9 [M + H].sup.+, C.sub.61H.sub.72O.sub.9N.sub.11Cl 568.7596; Found 568.7603 R.sub.T = 8.20 min (b) 49 b (HESI/FT) m/z: [M + 2H].sup.2+Calcd for B m/z 1095.1 [M + H].sup.+, C.sub.60H.sub.77O.sub.9N.sub.11 547.7947; Found 547.7954 R.sub.T = 7.25 min (b) 50 b (HESI/FT) m/z: [M + H].sup.+ Calcd for B m/z 1103.0 [M + H].sup.+, C.sub.59H.sub.74O.sub.9N.sub.11 1080.5666; Found 1080.5665 R.sub.T = 6.87 min (b) 51 b (HESI/FT) m/z: [M + H].sup.+ Calcd for B m/z 1136.9 [M + H].sup.+, C.sub.61H.sub.72O.sub.9N.sub.11Cl 568.7596; Found 568.7603 R.sub.T = 8.12 min (b) 52 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1110.1 [M + H].sup.+, C.sub.60H.sub.78O.sub.9N.sub.12 555.3002; Found 555.3022 R.sub.T = 7.27 min (b) 53 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1131.1 [M + H].sup.+, C611-17909N11 554.8026; Found 554.8026 R.sub.T = 7.33 min (b) 54 b (HESI/FT) m/z: [M + 2H].sup.+ Calcd for B m/z 1131.1 [M + H].sup.+, C.sub.61H.sub.79O.sub.9N.sub.11 554.8026; Found 554.8039 R.sub.T = 8.03 min (b) 55 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1132.0 [M + H].sup.+, C.sub.63H.sub.76O.sub.10N.sub.10 566.2868; Found 566.2881 R.sub.T = 7.71 min (b) 56 b (HESI/FT) m/z: [M + 2H].sup.2 +Calcd for B m/z 1144.0 [M + H].sup.+, C.sub.64H.sub.79O.sub.10N.sub.10 572.2868.; Found 572.2867 R.sub.T = 7.49 min (b) 57 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1139.0 [M + Na].sup.+, C.sub.62H.sub.76O.sub.9N.sub.11 558.7869; Found 558.7883 R.sub.T = 8.02 min (b) 58 b (HESI/FT) m/z: [M + 2H].sup.+ Calcd for m/z 1137.9 [M + H].sup.+, C.sub.60H.sub.71O.sub.9N.sub.12Cl 569.2572; Found 569.2586 B R.sub.T = 6.35 min (b) 59 b (HESI/FT) m/z: [M + H].sup.+ Calcd for B m/z 1137.9 [M + H].sup.+, C.sub.60H.sub.71O.sub.9N.sub.12Cl 569.2572; Found 569.2570 R.sub.T = 7.43 min (b) 60 b (HESI/FT) m/z: [M + H].sup.+ Calcd for B m/z 1116.9 [M + H].sup.+, C.sub.62H.sub.75O.sub.9N.sub.11 558.7869; Found 558.7869 R.sub.T = 7.03 min (b) 61 b (HESI/FT) m/z: [M + H].sup.+ Calcd for B m/z 1029.0 +M+30H+, C.sub.59H.sub.66O.sub.8N.sub.9 1028.5029; Found 1028.5056 R.sub.T = 10.48 min (b) 62 b (HESI/FT) m/z: [M + H].sup.+ Calcd for B m/z 1028.0 [M + H].sup.+, C.sub.60H.sub.67O.sub.8N.sub.8 1027.5076; Found 1027.5103 R.sub.T = 10.32 min (b) 63 b (HESI/FT) m/z: [M + H].sup.+ Calcd for B m/z 1080.9 [M + Na].sup.+, C.sub.60H.sub.68O.sub.9N.sub.9 1058.5135; Found 1058.5175 R.sub.T = 8.52 min (c) 64 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1104.9 [M + H].sup.+, C.sub.60H.sub.71O.sub.10N.sub.11 552.7687; Found 552.7687 R.sub.T = 7.35 min (b) 65 b (HESI/FT) m/z: [M + 2H].sup.2+Calcd for B m/z 514.8 [M + H].sup.+, C.sub.59H.sub.68O.sub.7N.sub.10 514.26307; Found 514.2651 R.sub.T = 8.31 min (b) 66 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1174.7 [M + H].sup.+, C.sub.65H.sub.78O.sub.11N.sub.10 587.2920; Found 587.2940 R.sub.T = 7.58 min (b) 67 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for m/z 1188.9 [M + H].sup.+, C.sub.60H.sub.71O.sub.9N.sub.11 544.7713; Found 544.7713 R.sub.T = 6.69 min (b) 68 b (HESI/FT) m/z: [M + H].sup.2+ Calcd for B m/z 1058.9 [[M + H].sup.+, C.sub.60H.sub.68O.sub.9N.sub.9 1058.5135; Found 1058.5135 R.sub.T = 10.71min (b) 69 b ND B m/z 1067.9 [M + 2H].sup.2+, R.sub.T = 8.61 min (b) 70 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 545.5 [M + 2H].sup.2+, C.sub.57H.sub.69O.sub.9N.sub.11F.sub.2 544.7618; Found 544.7618 R.sub.T = 7.16 min (b) 71 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1110.0 [M + 2H].sup.2+, C.sub.60H.sub.78O.sub.9N.sub.12 555.3002; Found 555.3002 R.sub.T = 7.03 min (b) 72 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 559.1 [M + 2H].sup.2+, C.sub.62H.sub.73O.sub.9N.sub.11 557.7791; Found 557.7804 R.sub.T = 7.92 min (b) 73 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 514.3 [M + 2H].sup.2+, C.sub.60H.sub.69O.sub.7N.sub.9 513.7655; Found 513.7675 R.sub.T = 8.48 min (b) 74 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1168.0 [M + Na].sup.+, C.sub.64H.sub.78O.sub.10N.sub.10 573.2946; Found 573.2966 R.sub.T = 8.02 min (b) 75 b (HESI/FT) m/z: [M + H].sup.+ Calcd for B m/z 1057. [M + H].sup.+, C.sub.61H.sub.69O.sub.9N.sub.8 1057.5182; Found 1057.5182 R.sub.T =10.79 min (b) 76 b (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for B m/z 1118.9 [M + H].sup.+, C.sub.62H.sub.74O.sub.10N.sub.10 559.2789; Found 559.2809 R.sub.T = 8.40 min (b) 77 b ND B m/z 1144.0 [M + H].sup.+, R.sub.T = 7.44 min (b) 78 b (HESI/FT) rn/z: [M + 2H].sup.2+ Calcd for B m/z 577.7 [M + 2H].sup.2+, C661-17809N10 577.2971; Found 577.2985 R.sub.T = 7.75 min (b) 79 a (HESI/FT) m/z: [M + 2H].sup.2+ Calcd for C m/z 1132.3 [M + H].sup.+, C.sub.63H.sub.73O.sub.9N.sub.10F 566.2695; Found 566.2785 R.sub.T = 8.75 min ND—Not determined
TABLE-US-00005 TABLE 3 .sup.1H NMR and .sup.13C NMR analysis for selected Examples Exa mple Structure .sup.1H NMR .sup.13C NMR 3
Biological Activity
[0186] A Hit Hunter® cAMP XS+ assay was used to monitor activation of the SST.sub.2 and SST.sub.5 via the Gi G protein. Cells were pre-stimulated with forskolin to elicit a cAMP response and the potency and efficacy of somatostatin agonists to reduce cAMP was measured. Briefly cells (10,000 cells/well) were incubated (37° C.; 30 min) with compound (10 μM-10 fM) in media (2:1 HBSS/10 mM Hepes:cAMP XS+ Ab reagent) in the presence of EC.sub.80 forskolin (20 μM SST.sub.2 and 15 μM SST.sub.5). Signal was detected through incubation with 20 μL cAMP XS+ ED/CL lysis cocktail (60 min; RT)
[0187] followed by incubation with 20 μL cAMP XS+ EA reagent (180 min; RT). Microplates were read with a PerkinElmer Envision™ instrument for chemiluminescent signal detection. Compound activity was normalised to the activity of somatostatin 28 (basal 0%, 100%=signal achieved with 50 nM somatostatin 28) and data analyzed using Dotmatics Browser using a 4-parameter logistic fit to determine potency (pEC.sub.50) and efficacy (%).
TABLE-US-00006 SST.sub.5/2 Example Number SST.sub.2 pEC.sub.50 SST.sub.2 (EMax) SST.sub.5 pEC.sub.50 SST.sub.5 (EMax) Selectivity Somatostatin-28 9.6 100.4 10.3 101.0 4.6 Pasireotide 9.0 100.3 9.6 101.5 3.8 Octreotide 10.5 98.5 7.8 97.0 0.0 Example 1 5.5 62.0 7.6 98.6 130.7 Example 2 6.4 96.4 7.7 104.8 21.9 Example 3 6.7 98.4 9.1 100.0 236.7 Example 4 6.6 102.9 7.9 105.0 23.6 Example 5 6.5 97.6 8.0 98.7 33.8 Example 6 7.3 97.9 8.9 97.9 41.9 Example 7 8.0 99.2 9.3 101.5 19.3 Example 8 7.5 101.2 8.9 102.0 22.7 Example 9 7.3 101.5 8.6 106.0 23.3 Example 10 7.7 100.8 9.3 98.0 38.9 Example 11 6.7 98.0 8.4 99.4 48.2 Example 12 7.7 97.6 9.1 98.1 26.1 Example 13 6.7 97.6 8.6 98.7 87.2 Example 14 6.8 99.6 10.1 96.7 1876.8 Example 15 <5.0 55.5 7.7 100.7 531.3 Example 16 <5.0 79.5 8.4 100.5 2729.5 Example 17 6.4 97.3 9.3 103.8 789.4 Example 18 8.3 28.2 7.9 100.1 0.3 Example 19 7.0 75.5 7.7 99.8 5.5 Example 20 5.5 98.0 8.2 99.6 418.4 Example 21 6.4 97.3 8.7 101.1 220.3 Example 22 7.1 100.4 9.3 101.7 142.6 Example 23 6.2 96.6 8.4 100.7 148.7 Example 24 <5 46.8 7.0 101.7 110.6 Example 25 6.7 100.3 9.9 102.4 1351.8 Example 26 6.3 66.6 9.9 99.1 4067.3 Example 27 6.4 90.7 8.6 99.4 159.6 Example 28 8.1 99.6 10.1 100.4 101.0 Example 29 6.1 89.6 9.0 100.0 663.6 Example 30 6.6 100.1 9.0 101.4 214.2 Example 31 6.7 99.7 10.2 88.7 3441.5 Example 32 7.0 98.8 8.9 99.9 66.8 Example 33 <5.0 13.4 7.4 107.2 279.1 Example 34 5.4 49.2 9.0 101.9 3271.8 Example 35 <5.0 27.8 9.0 107.2 10046.0 Example 36 6.3 63.4 8.9 101.9 384.3 Example 37 5.2 75.8 8.7 107.5 2927.5 Example 38 5.6 82.7 7.9 103.1 171.5 Example 39 6.4 100.3 9.4 106.6 1104.6 Example 40 6.5 91.2 8.6 105.7 126.6 Example 41 8.1 102.3 9.8 104.0 51.0 Example 42 7.9 99.3 9.6 104.0 45.1 Example 43 7.5 101.3 9.5 106.7 94.3 Example 44 6.1 75.3 9.0 105.1 823.8 Example 45 6.8 100.5 9.4 100.1 430.7 Example 46 7.4 98.6 10.1 101.5 527.6 Example 47 6.7 100.1 9.1 108.0 226.4 Example 48 8.7 100.3 10.7 102.8 106.6 Example 49 6.3 98.9 8.5 103.3 143.3 Example 50 6.2 95.4 7.8 103.3 41.1 Example 51 8.3 100.3 10.1 106.4 64.3 Example 52 6.4 96.8 9.8 107.4 2108.2 Example 53 <5.0 50.9 7.5 102.7 293.1 Example 54 6.2 96.2 9.3 104.0 1083.5 Example 55 <5 85.9 8.0 106.4 1120.0 Example 56 6.5 98.1 9.7 104.3 1648.2 Example 57 6.7 96.5 9.3 102.1 404.6 Example 58 <5.0 42.9 7.8 103.0 695.7 Example 59 6.3 95.7 9.2 106.2 738.9 Example 60 6.9 99.8 9.0 104.3 129.4 Example 61 <5.0 23.4 7.5 103.3 291.4 Example 62 6.2 99.4 8.5 101.8 224.3 Example 63 7.2 102.4 8.6 103.8 24.4 Example 64 7.0 100.3 7.8 103.8 5.5 Example 65 6.0 91.8 9.4 99.4 2293.7 Example 66 <5.0 51.1 8.9 102.7 7454.5 Example 67 6.4 98.3 7.8 103.7 25.7 Example 68 6.6 99.1 9.5 98.8 791.6 Example 69 <5.0 69.0 7.1 102.7 124.3 Example 70 <5.0 77.9 8.3 102.2 1815.8 Example 71 <5.0 52.8 7.2 103.2 171.4 Example 72 <5.0 14.0 8.2 101.3 1621.3 Example 73 6.4 99.2 9.2 94.9 566.7 Example 77 5.4 39.3 8.2 103.6 681.8 Example 78 5.5 76.2 8.9 105.5 2726.7