HK1110313B - Stilbene derivatives and their use for binding and imaging amyloid plaques - Google Patents

Description

Stilbene derivatives and their use for binding and imaging amyloid plaques

The present application is a divisional application of chinese patent application, chinese patent application 02816829.1 filed on 8/27/2002, entitled "stilbene derivatives and their use for binding and imaging amyloid plaques".

Technical Field

The present invention relates to novel biologically active compounds, methods of diagnostic imaging using emittance labeled compounds, and methods of preparing emittance labeled compounds.

Background

Alzheimer's Disease (AD) is a progressive neurodegenerative disorder characterized by cognitive decline, irreversible memory loss, disorientation, and language impairment. Post-mortem examination of AD brain sections demonstrated a large number of Senile Plaques (SP) composed of amyloid- β (A β) peptides and a number of neurofibrillary clusters (NFTs) formed by filaments of highly phosphorylated tau (recent reviews and other references see Ginsberg, S.D. et al, "Molecular Pathology of Alzheimer's Disease and Related Disorders," in cellular Cortex: neuro-generative and agent-Related Changes in structural function of cellular Cortex, Kluwer Academic/Plenum, NY 1999, p.603-654, Vogelberg-Ragag, V.et al, "Cell Biology of genetic and cellular Pathology in Alzheimer's Disease," diseases of Alzheimer's Disease, "P.359, Wilford's Disease, P.372). Family AD (FAD) is caused by multiple mutations in the A Precursor Protein (APP), presenilin 1(PS1) and presenilin 2(PS2) genes (Ginsberg, S.D., et al, "molecular Pathology of Alzheimer's Disease and Related Disorders," InCerea syndrome: Neodygegenous and Age-Related Changes in Structure and Function of Cerea syndrome, KluwerAcademic/plunger, NY (1999), pp.603-654; Vogelberg-Ragagliia, V.et al, "Biology of Tau and cytokine pathway in Alzheimer's Disease," Alzheimer's Disease, P.372, William. P.359, P.1999).

Although the precise mechanism of the underlying AD is not fully understood, all the pathogenic FAD mutations studied in this way resulted in a large increase in more amyloid-producing 42-43 amino acid long forms of a β peptide. Thus, at least in FAD, dysregulation of a β production appears to be sufficient to induce a cascade of events leading to neurodegeneration. In fact, this amyloid cascade hypothesis suggests that extracellular fibrillar A.beta.aggregates formed in the brain may be a key event in the pathogenesis of AD (Selkoe, D.J., "Biology of B-amyloid prefocutor Protein and the mechanism of Alzheimer's Disease," Alzheimer's Disease, Lippincot Williams & Wilkins, Philadelphia, PA (1999), pp.293-310; Selkoe, D.J.; J.Am. Med. Ashoc.283: 1615-.

Various protocols that attempt to inhibit the production of fibers A.beta.in the brain and induce aggregation thereof are currently being evaluated as potential therapies for AD (Skovronsky, D.M. and Lee, V.M., TrendsPharmacol. Sci.21: 161-163 (2000); Vassar, R., et al, Science 286: 735-Chem 741 (1999); Wolfe, M.S. et al, J. Med.Chem.41: 6-9 (1998); Moore, C.L. et al, J. Med.Chem.43: 3434-3442 (2000); Findeis, M.A., Biochimica Acta 1502: 76-84 (2000); Kuner, P., Bohrmann et al, J.biol.Chem.275: 3-1678 (2000)). There is therefore a great interest in developing ligands that specifically bind to fibrillar a β aggregates. Since extracellular SPs are accessible targets, these novel ligands can be used as in vivo diagnostic tools and as probes in studies of AD amyloidogenesis in living patients to visualize progressive deposition of a β.

For this reason, several schemes for the development of fibrillar A β aggregate-specific ligands have been reported (Ashburn, T.T. et al, chem.biol.3: 351-358 (1996); Han, G. et al, J. Am. chem. Soc. 118: 4506-4507 (1996); Klunk, W.E. et al, biol.Psychiatry 35: 627 (1994); Klunk, W.E. et al, Neuroobiol.Aging 16: 541-548 (1995); Klunk, W.E. et al, Society for Neurospora extract 23: 1638 (1997); Mathis, C.A. et al, Proc.XIh Intl. Symp. radior. chem. uppsalm, Uppsala, Sweden: 94-95; Lonrez A. 1997, J. Acad. A. et al, Nat. Zhak.5. J. 1985: 12242, Nat. Zhak. 5. J. 1985, Nat. S. 10. 19847, Nature). The most attractive approach is based on highly conjugated Huangfenning-G (CG) and Congo Red (CR), and the latter has been used for fluorescent staining of SPs and NFTs in post-mortem AD brain sections (Ashburn, T.T. et al, chem.biol. 3: 351-358 (1996); Klunk, W.E. et al, J.Histochem. Cytochem. 37: 1273-1281 (1989)). The inhibition constants (Ki) for binding to fibrillar A β aggregates of CR, CG and the 3 '-bromo-and 3' -iodo-derivatives of CG were 2,800, 370, 300 and 250nM, respectively (Mathis, C.A. et al, Proc. XIIth Intl. Sump. radiopharm. chem., Uppsa1a, Sweden: 94-95 (1997)). These compounds have been shown to bind selectively to A β (1-40) peptide aggregates in vitro and to fibrous A β deposits in AD brain sections (Mathis, C.A. et al, Proc. XIIth Intl. Symp. radiopharmam. chem., Uppsala, Sweden: 94-95 (1997)).

Amyloid disorders are characterized by the accumulation of various insoluble fibrous proteins within the tissues of a patient. Amyloid deposits are formed by aggregating amyloid proteins, followed by further binding of the aggregates and/or amyloid proteins. The formation and aggregation of aggregates of β -amyloid (a β) peptide in the brain is a key factor in the presentation and development of AD. Fibrillar aggregates of these amyloid proteins, A β_1-40And Abeta_1-42The major metabolic peptides derived from amyloid precursor protein found in senile plaques and cerebrovascular amyloid deposits in AD patients (Xia, W. et al, J.Proc. Natl. Acad. Sci.U.S.A. 97: 9299-9304 (2000)). Prevention and reversal of AB plaque formation has long been the target for treatment of this disease (Selkoe, D., J.JAMA283: 1615-1617 (2000); Wolfe, M.S. et al, J.Med. chem. 41: 6-9 (1998); Skovronsky, D.M. and Lee, V.M., Trends Pharmacol. Sc. (Sc); Selkoe, D.D., J.JAMA283: 1615-1617; Wolfe, M.S. et al, J.Med. chem. 41: 6-9(1998)i.21：161-163(2000))。

In addition to the role of amyloid deposits in alzheimer's disease, the presence of amyloid deposits has been shown in the following diseases: mediterranean fever, Muckle-Wells syndrome, idiopathic myeloma, amyloid polyneuropathy, amyloid cardiomyopathy, systemic senile amyloid disease, amyloid polyneuropathy, hereditary cerebral hemorrhage with amyloidosis, Down's syndrome, scrapie, Creutzfeldt-Jacob disease, kuru, gerstmann-Straussler-Scheinker syndrome, bone marrow cancer of the thyroid gland, isolated atrial amyloid, beta-amyloid in dialysis patients₂Beta in microglobulin amyloid, inclusion body myositis, muscle wasting diseases₂Amyloid deposits and insulinoma of type II diabetes.

Therefore, a simple and non-invasive method for detecting and quantifying amyloid deposits in patients is urgently sought. Currently, the detection of amyloid deposits involves histological analysis of biopsy or autopsy material. Both of these methods have drawbacks. For example, autopsy can only be used for post mortem diagnosis.

The imaging agent may be based on two isotopes.^99mTc(T_1/26 hours; 140KeV) and¹²³I(T_1/213 hours; 159KeV) is commonly used for Single Photon Emission Computed Tomography (SPECT), and¹¹C(T_1/220 minutes; 511KeV) and¹⁸F(T_1/2110 minutes; 511KeV) is commonly used for Positron Emission Tomography (PET).

Since amyloid deposits have many of the same physical properties (e.g., density and moisture content) as normal tissue, these deposits are difficult to directly image in vivo. Attempts to image amyloid deposits using Magnetic Resonance Imaging (MRI) and computer-assisted tomography (CAT) have been disappointing and have detected amyloid deposits only under certain favorable conditions. Furthermore, attempts to label amyloid deposits with antibodies, serum amyloid P protein, or other probe molecules have provided some selectivity for peripheral tissues, but provide aberration within the tissue.

Potential ligands for detecting Α β aggregates in living brains must cross the entire blood brain barrier. The use of relatively small molecule (compared to congo red) ligands can therefore improve brain uptake and increase lipophilicity. Highly conjugated thioflavins (S and T) are commonly used as dyes for staining A.beta.aggregates in AD brain (Elhaddaoui, A. et al, biospectropy 1: 351-356 (1995)). These compounds are predominantly benzothiazole-based, having a relatively small molecular size.

A non-invasive technique for imaging and quantifying amyloid deposits in a patient would be useful. In addition, it would be useful to provide compounds that inhibit amyloid aggregation to form amyloid deposits and a method of detecting the ability of a compound to inhibit amyloid aggregation.

Summary of The Invention

The present invention provides novel compounds of formula I, II, III, IV or V.

The invention also provides diagnostic compositions comprising an emittance labeled compound of formula I, II, III, IV or V and a pharmaceutically acceptable carrier or diluent.

The invention also provides a method of imaging amyloid deposits comprising introducing into a patient a detectable amount of a labeled compound of formula I, II, III, IV, or V, or a pharmaceutically acceptable salt, ester, amide, or prodrug thereof.

The invention also provides a method of inhibiting amyloid aggregation comprising administering to a mammal an amyloid inhibiting amount of a compound of formula I, II, III, IV or V or a pharmaceutically acceptable salt, ester, amide or prodrug.

Another aspect of the invention relates to processes and intermediates for synthesizing the amyloid inhibiting and imaging compounds of formula I, II, III, IV or V described herein.

Brief Description of Drawings

Figures 1, 3, 4 and 5 depict representative compounds of the invention and binding data for these compounds.

FIG. 2 depicts binding data for compounds of the invention.

Detailed Description

A first aspect of the invention relates to compounds of formula I:

or a pharmaceutically acceptable salt thereof, wherein:

R⁵is hydrogen or C_1-4An alkyl group;

in each case, R¹、R²And R³Independently selected from the group consisting of: hydrogen, halogen, C_1-4Alkyl, cyano, carboxy (C)_1-5) Alkyl, trifluoromethyl, nitro, methylamino, dimethylamino, halo (C)_1-4) Alkyl and formyl;

R⁴selected from the group consisting of:

a.C_1-4an alkylthio group is a group of one or more,

b. halogen (C)_1-4) An alkoxy group,

c. carboxy (C)_1-5) An alkyl group, a carboxyl group,

d. a hydroxyl group(s),

e.C_1-4an alkoxy group,

f.NR⁶R⁷wherein

R⁶And R⁷Is hydrogen, halogen (C)_1-4) Alkyl or C_1-4An alkyl group, a carboxyl group,

g. phenyl (C)_1-4) An alkyl group, a carboxyl group,

h.C_6-10an aryl group, a heteroaryl group,

i. (ii) a heteroaryl group, wherein,

j. a heterocyclic ring,

k. heterocycle (C)_1-4) Alkyl, and

l.C_3-6a cycloalkyl group,

wherein said phenyl group (C)_1-4) Alkyl radical, C_6-10Aryl, heteroaryl, heterocycle (C)_1-4) Alkyl or C_3-6Cycloalkyl is substituted with one of the following substituents: c_1-4Alkylthio radical, C_1-4Alkylsulfonyl, methoxy, hydroxy, dimethylamino, or methylamino;

and the combination of (a) and (b),

x' is¹²⁵I、¹²³I、¹³¹I、¹⁸F、¹⁸Fluorine (C)_1-4) Alkyl group, [2 ]¹⁸Fluorine (C)_1-4) Alkyl radical]Alkylamino radical, [2 ]¹⁸Fluorine (C)_1-4) Alkyl radical]Amino group,⁷⁶Br、⁷⁷Br or Sn (alkyl)₃。

Useful compounds falling within the scope of formula I include those wherein R⁵Is hydrogen or C_1-4Alkyl compounds. R⁵Particularly useful values of (a) are hydrogen and methyl. R⁵The most useful value of (a) is hydrogen.

Useful compounds are those in which in each case R¹、R²And R³A compound of formula I independently selected from the groups as described above. Preferably, R³Is hydrogen. In this preferred embodiment, R is particularly preferred¹And R²Independently selected from the group consisting of: hydrogen and C_1-4An alkyl group. More preferably, R¹And R²At least one of which is hydrogen. Most preferably, R¹And R²Are all hydrogen.

Useful compounds of formula I also include those wherein R⁴Such as those described above. R⁴At C_6-10Preferred values in the range of aryl include phenyl, naphthyl or tetrahydronaphthyl. R⁴Preferred values in the range of heteroaryl groups include thienyl, furyl, pyranyl, pyrrolyl, pyridyl, indolyl and imidazolyl. R⁴Preferred values within the scope of the heterocyclic ring include piperidinyl, pyrrolidinyl and morpholinyl. In which R is⁴Is C_6-10Aryl, heteroaryl, heterocycle (C)_1-4) Alkyl or C_3-6In compounds of one preferred embodiment of cycloalkyl, most preferably the ring is substituted with one of the following substituents: c_1-4Alkylthio, carboxyl (C)_1-5) Alkyl, hydroxy, methoxy, dimethylamino, or methylamino. In another embodiment, R⁴More preferably selected from the group consisting of: c_1-4Alkylthio, halogen (C)_1-4) Alkoxy, carboxyl (C)_1-5) Alkyl, hydroxy, C_1-4Alkoxy and NR⁶R⁷Wherein R is⁶And R⁷Independently of each other hydrogen, halogen (C)_1-4) Alkyl or C_1-4An alkyl group. Most preferably, R⁴Selected from the group consisting of: methylthio, carboxymethyl, carboxyethyl, carboxypropyl, hydroxy, methoxy or NR⁶R⁷Wherein R is⁶And R⁷Independently of each other hydrogen, fluorine (C)_1-4) Alkyl or methyl.

Useful values of X' include¹²⁵I、¹²³I、¹³¹I、¹⁸F、¹⁸Fluorine (C)_1-4) Alkyl group, [2 ]¹⁸Fluorine (C)_1-4) Alkyl radical]Alkylamino radical, [2 ]¹⁸Fluorine (C)_1-4) Alkyl radical]Amino group,⁷⁶Br、⁷⁷Br or Sn (alkyl)₃. Particularly useful values of X' are¹²³I、¹⁸A fluoromethyl group,¹⁸Fluoroethyl and¹⁸and (3) fluoropropyl.

The invention also relates to compounds of formula II:

or a pharmaceutically acceptable salt thereof,

z is O, S or NR^aWherein

R^aIs C_1-4An alkyl group;

in each case, R⁹、R¹⁰And R¹¹Independently selected from the group consisting of: hydrogen, halogen, C_1-4Alkyl, cyano, carboxy (C)_1-5) Alkyl, trifluoromethyl, nitro, methylamino, dimethylamino, halo (C)_1-4) Alkyl and formyl;

R¹²selected from the group consisting of:

a.C_1-4an alkylthio group is a group of one or more,

b. halogen (C)_1-4) An alkoxy group,

c. carboxy (C)_1-5) An alkyl group, a carboxyl group,

d. a hydroxyl group(s),

e.C_1-4an alkoxy group,

f.NR¹³R¹⁴wherein

R¹³And R¹⁴Is hydrogen, halogen (C)_1-4) Alkyl or C_1-4An alkyl group, a carboxyl group,

g. phenyl (C)_1-4) An alkyl group, a carboxyl group,

h.C_6-10an aryl group, a heteroaryl group,

i. (ii) a heteroaryl group, wherein,

j. a heterocyclic ring,

k. heterocycle (C)_1-4) Alkyl, and

l.C_3-6a cycloalkyl group,

wherein said phenyl group (C)_1-4) Alkyl radical, C_6-10Aryl, heteroaryl, heterocycle (C)_1-4) Alkyl or C_3-6Cycloalkyl is substituted with one of the following substituents: c_1-4Alkylthio radical, C_1-4Alkylsulfonyl, methoxy, hydroxy, dimethylamino, or methylamino;

and the combination of (a) and (b),

x' is¹²³I、¹²³I、¹³¹I、¹⁸F、¹⁸Fluorine (C)_1-4) Alkyl group, [2 ]¹⁸Fluorine (C)_1-4) Alkyl radical]Alkylamino radical, [2 ]¹⁸Fluorine (C)_1-4) Alkyl radical]Amino group,⁷⁶Br、⁷⁷Br or Sn (alkyl)₃。

Useful compounds falling within the scope of formula II include those wherein Z is O, S or NR^aWherein R is^aIs C_1-4An alkyl group. Particularly useful compounds are those wherein Z is O.

Useful compounds are those in which in each case R⁹、R¹⁰And R¹¹A compound of formula I independently selected from the groups as described above. Preferably, R¹¹Is hydrogen. In this preferred embodiment, R is particularly preferred⁹And R¹⁰Independently selected from the group consisting of: hydrogen and C_1-4An alkyl group. More preferably, R⁹And R¹⁰At least one of which is hydrogen. Most preferably, R⁹And R¹⁰Are all hydrogen.

Useful compounds of formula I also include those wherein R¹²Such as those described above. R¹²At C_6-10Preferred values in the range of aryl include phenyl, naphthyl or tetrahydronaphthyl. R¹²Preferred values in the range of heteroaryl groups include thienyl, furyl, pyranyl, pyrrolyl, pyridyl, indolyl and imidazolyl. R¹²Preferred values within the scope of the heterocyclic ring include piperidinyl, pyrrolidinyl and morpholinyl. At itIn R¹²Is C_6-10Aryl, heteroaryl, heterocycle (C)_1-4) Alkyl or C_3-6In compounds of one preferred embodiment of cycloalkyl, most preferably the ring is substituted with one of the following substituents: c_1-4Alkylthio, carboxyl (C)_1-5) Alkyl, methoxy, hydroxy, dimethylamino, or methylamino. In another embodiment, R¹²More preferably selected from the group consisting of: c_1-4Alkylthio, halogen (C)_1-4) Alkoxy, carboxyl (C)_1-5) Alkyl, hydroxy, C_1-4Alkoxy and NR¹³R¹⁴Wherein R is¹³And R¹⁴Independently of each other hydrogen, halogen (C)_1-4) Alkyl or C_1-4An alkyl group. Most preferably, R¹²Selected from the group consisting of: methylthio, carboxymethyl, carboxyethyl, carboxypropyl, hydroxy, methoxy or NR¹³R¹⁴Wherein R is¹³And R¹⁴Independently of each other hydrogen, fluorine (C)_1-4) Alkyl or methyl.

Useful values of X' include¹²⁵I、¹²³I、¹³¹I、¹⁸F、¹⁸Fluorine (C)_1-4) Alkyl group, [2 ]¹⁸Fluorine (C)_1-4) Alkyl radical]Alkylamino radical, [2 ]¹⁸Fluorine (C)_1-4) Alkyl radical]Amino group,⁷⁶Br、⁷⁷Br or Sn (alkyl)₃. Particularly useful values of X' are¹²³I、¹⁸A fluoromethyl group,¹⁸Fluoroethyl and¹⁸and (3) fluoropropyl.

Another aspect of the invention relates to a compound of formula III:

or a pharmaceutically acceptable salt thereof, wherein:

n is equal to a value of 0 to 4,

R²⁸is hydrogen or C_1-4An alkyl group, a carboxyl group,

z is O, S or CR¹⁵＝CR¹⁶-, wherein

R¹⁵、R¹⁶、R¹⁷、R¹⁸、R¹⁹、R²⁰、R²¹、R²²、R²³、R²⁴And R²⁵In each case independently selected from the group consisting of: hydrogen, halogen, Sn (alkyl)₃、C_1-4Alkyl radical, C_1-4Alkylsulfanyl group, C_1-4Alkylsulfonyl radical, C_1-4Alkoxy, hydroxy, C_6-10Aryl, carboxyalkyl, carboxyl and NR²⁶R²⁷Wherein

R²⁶And R²⁷Independently of each other is hydrogen, C_1-4Alkyl, phenyl (C)_1-4) Alkyl, halo (C)_1-4) Alkyl, haloaryl (C)_1-4) Alkyl radical, C_6-10Aryl, heteroaryl, heterocycle (C)_1-4) Alkyl or C_3-6Cycloalkyl radicals, in which

Said C is_6-10Aryl radical, C_6-10Heteroaryl, heterocycle or C_3-6Cycloalkyl is unsubstituted or substituted with one of the following substituents: c_1-4Alkylthio radical, C_1-4Alkylsulfonyl, methoxy, hydroxy, dimethylamino or methylamino,

And the combination of (a) and (b),

R^Pis hydrogen or a sulphur-protecting group, for example methoxymethyl, methoxyethoxymethyl, p-methoxybenzyl or benzyl.

The tetradentate metal-ligand moiety of formula III is capable of complexing with a metal, for example 99 m-pertechnetate, to form a metal chelate as described herein, for example of the formula:

additionally, a rhenium-emitting isotope may be complexed with the tetradentate metal ligand.

Useful compounds of formula III are those wherein Z is O, S or-CR¹⁵＝CR¹⁶Those compounds of (a) wherein R is¹⁵And R¹⁶As described above. Preferably, Z is-CR¹⁵＝CR¹⁶-, wherein R¹⁵And R¹⁶As described above. More preferably, R¹⁵And R¹⁶Are all hydrogen.

Useful compounds of the invention are those wherein R is¹⁷To R²⁵All as defined above. R¹⁷To R²⁵Falls into C_6-10Preferred values in the range of aryl include phenyl, naphthyl or tetrahydronaphthyl. R¹⁷To R²⁵Preferred values falling within the scope of heteroaryl include thienyl, furyl, pyranyl, pyrrolyl, pyridyl, indolyl and imidazolyl. R¹⁷To R²⁵Preferred values falling within the scope of heterocycles include piperidinyl, pyrrolidinyl and morpholinyl. At R¹⁷To R²⁵Is C_6-10Aryl, heteroaryl, heterocycle (C)_1-4) Alkyl or C_3-6In compounds of one preferred embodiment of cycloalkyl, most preferably the ring is substituted with one of the following substituents: c_1-4Alkylthio radical, C_1-4Alkylsulfonyl, methoxy, hydroxy, dimethylamino, or methylamino. In another embodiment, more preferred compounds include those wherein R is¹⁷To R²⁵Those in which one or more is hydrogen. In this embodiment, R is preferred¹⁷Is not hydrogen. More preferably, R¹⁷Selected from the group consisting of: c_1-4Alkylthio radical, C_1-4Alkylsulfonyl, hydroxy, C_1-4Alkoxy, NR²⁶R²⁷Wherein R is²⁶And R²⁷Independently is hydrogen or C_1-4An alkyl group. Most preferably, R¹⁷Is NR²⁶R²⁷Wherein R is²⁶And R²⁷Are both methyl groups.

Useful compounds also include those of formula III wherein n is equal to a value of 0 to 4. Preferably, n is equal to a value of 0-2. More preferably, n is equal to 0.

Yet another aspect of the invention relates to a compound of formula IV:

or a pharmaceutically acceptable salt thereof, wherein

n is equal to a value of 0 to 4,

R²⁹、R³⁰、R³¹、R³²、R³³、R³⁴and R³⁵Independently selected from the group consisting of:

a. the presence of hydrogen in the presence of hydrogen,

b.C_1-4an alkylthio group is a group of one or more,

c.C_1-4an alkyl sulfonyl group, a carboxyl group,

d. a hydroxyl group(s),

e.C_1-4an alkoxy group,

f.NR⁶R⁷wherein

R⁶And R⁷Is hydrogen or C_1-4An alkyl group, a carboxyl group,

g. phenyl (C)_1-4) An alkyl group, a carboxyl group,

h.C_6-10an aryl group, a heteroaryl group,

i. (ii) a heteroaryl group, wherein,

j. a heterocyclic ring,

k. heterocycle (C)_1-4) Alkyl, and

l.C_3-6a cycloalkyl group,

wherein said phenyl group (C)_1-4) Alkyl radical, C_6-10Aryl, heteroaryl, heterocycle (C)_1-4) Alkyl or C_3-6Cycloalkyl is substituted byA substitution of: c_1-4Alkylthio radical, C_1-4Alkylsulfonyl, methoxy, hydroxy, dimethylamino, or methylamino;

provided that R is²⁹To R³⁵One is a monoalkylaminophenyl or dialkylaminophenyl group; and

R^Pis hydrogen or a sulphur protecting group, such as methoxymethyl, methoxyethoxymethyl, p-methoxybenzyl or benzyl.

Useful compounds of formula IV are those wherein R²⁹、R³⁰、R³¹、R³²、R³³、R³⁴And R³⁵Such as those described above. R²⁹To R³⁵Falls into C_6-10Preferred values in the range of aryl include phenyl, naphthyl or tetrahydronaphthyl. R²⁹To R³⁵Preferred values falling within the scope of heteroaryl include thienyl, furyl, pyranyl, pyrrolyl, pyridyl, indolyl and imidazolyl. R²⁹To R³⁵Preferred values falling within the scope of heterocycles include piperidinyl, pyrrolidinyl and morpholinyl. In which R is²⁹To R³⁵Is C_6-10Aryl, heteroaryl, heterocycle (C)_1-4) Alkyl or C_3-6In the compounds of the preferred embodiments in cycloalkyl, most preferably the ring is substituted with one of the following substituents: c_1-4Alkylthio radical, C_1-4Alkylsulfonyl, methoxy, hydroxy, dimethylamino, or methylamino. In another embodiment, particularly useful compounds are those wherein R is²⁹、R³⁰、R³¹And R³³All of which are hydrogen. In this embodiment, R is particularly preferred³²And³⁴one of which is as described above, R³²And R³⁴The other is hydrogen. More preferably, R³²And R³⁴One of which is aminophenyl, monoalkylaminophenyl or dialkylaminophenyl, R³²And R³⁴The other is hydrogen. Most preferably, R³²And R³⁴One of them is a di-dimethylaminophenyl radical, R³²And R³⁴The other is hydrogen. R³⁵Useful values of also include hydrogen, methoxy, C_1-4Alkylthio radical, C_1-4Alkylsulfonyl, hydroxy and C_1-4An alkyl group. Most preferably, R³⁵Is hydrogen or C_1-4An alkyl group.

Useful compounds of formula IV also include compounds wherein n is equal to a value of 0 to 4. More preferably, n is equal to 0 or 1. Most preferably, n is equal to 0.

It is also to be understood that the present invention contemplates the inclusion of stereoisomers as well as optical isomers, such as mixtures of optical enantiomers as well as individual optical enantiomers and diastereomers, which arise as a result of structural asymmetry in the series of compounds described.

The compounds of formula I, II, III or IV may also be solvated, especially hydrated. Hydration occurs during the preparation of the compounds or compositions comprising the compounds, or may occur over time due to the hygroscopic nature of the compounds. In addition, the compounds of the present invention may exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. In general, the solvated forms are considered equivalent to unsolvated forms for the purposes of the present invention.

Yet another aspect of the invention relates to a compound of formula V:

or a pharmaceutically acceptable salt thereof or a derivative of a compound of formula V containing an emitting isotope complex, wherein:

r is C_1-4Alkyl or as above R²⁹-R³⁵Is defined by, and

R^Pas defined above.

When any change occurs more than once in any constituent or in formula I, II, III, IV or V, the definition in each case is independent of the definition in each other case. Likewise, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

The term "alkyl" as used herein by itself or as part of another group refers to straight or branched chain radicals of up to 8 carbons, preferably 6 carbons, more preferably 4 carbons, such as methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, and isobutyl.

The term "alkoxy" as used herein refers to a straight or branched chain alkyl radical attached to an oxygen atom, as described above, and includes, but is not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, and the like, unless the chain length is limited thereto. Preferably the alkoxy chain length is 1 to 6 carbon atoms, more preferably the chain length is 1 to 4 carbon atoms.

The term "monoalkylamine" as used herein by itself or as part of another group refers to an amino group substituted with one alkyl group as described above.

The term "dialkylamine" as employed herein by itself or as part of another group refers to an amino group substituted with two alkyl groups as described above.

The term "halo" as used herein by itself or as part of another group refers to chloro, bromo, fluoro or iodo.

The term "haloalkyl" as used herein refers to any of the above alkyl groups substituted with one or more of chloro, bromo, fluoro or iodo, preferably with fluoro and chloro, such as chloromethyl, iodomethyl, trifluoromethyl, 2,2, 2-trifluoroethyl and 2-chloroethyl.

The term "alkylthio" as used herein by itself or as part of another group refers to a thioether structure: R-S, wherein R is C as defined above_1-4An alkyl group.

The term "alkylsulfonyl" as used herein by itself or as another groupAnd a moiety refers to the sulfone structure: R-SO₂Wherein R is C as defined above_1-4An alkyl group.

The term "aryl" as employed herein by itself or as part of another group refers to a monocyclic or bicyclic aromatic group containing 6 to 12 carbons in the ring portion, preferably 6 to 10 carbons in the ring portion, such as phenyl, naphthyl or tetrahydronaphthyl.

The term "heterocycle" or "heterocyclic ring" as used herein, unless otherwise indicated, represents a stable 5-to 7-membered monocyclic heterocyclic ring system which may be saturated or unsaturated and which consists of carbon atoms and 1-3 heteroatoms selected from N, O, and S, and wherein the nitrogen and sulfur heteroatoms may optionally be oxidized. Especially useful are rings containing one nitrogen and one oxygen or sulfur or two nitrogen heteroatoms. Examples of such heterocyclic groups include piperidinyl, pyrrolyl, pyrrolidinyl, imidazolyl, imidazinyl, imidazolidinyl, pyridinyl, pyrazinyl, pyrimidinyl, imidazolyl, pyridyl, etc,Azolyl group,Oxazolidinyl, isoAzolyl radical, isoOxazolidinyl, thiazolyl, thiazolidinyl, isothiazolyl, homopiperidinyl, homopiperazinyl, pyridazinyl, pyrazolyl and pyrazolidinyl, most preferably thiomorpholinyl, pyrazinyl and morpholinyl.

The term "heteroatom" as used herein refers to an oxygen atom ("O"), a sulfur atom ("S"), or a nitrogen atom ("N"). It is contemplated that when the heteroatom is nitrogen, it may form NR^aR^bMoiety wherein R^aAnd R^bIndependently of one another, hydrogen or C_1-4Alkyl radical, C_2-4Aminoalkyl radical, C_1-4Haloalkyl, halobenzyl, or R¹And R²Together form a 5-to 7-membered heterocyclic ring optionally having O, S or NR in said ring^cWherein R is^cIs hydrogen or C_1-4An alkyl group.

The term "heteroaryl" as used herein refers to a group: they have 5 to 14 ring atoms; 6, 10 or 14 pi electrons shared in a ring array; and containing carbon atoms and 1, 2 or 3 oxygen, nitrogen or sulfur heteroatoms (examples of heteroaryl groups therein are thienyl, benzo [ b ]]Thienyl, naphtho [2, 3-b ]]Thienyl, thianthryl, furyl, pyranyl, isobenzofuryl, benzofuranylAzolyl, chromenyl, xanthenyl, benzothiophenyl (phenoxathiinyl), 2H-pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, indazolyl, purinyl, 4H-quinolizinyl, isoquinolyl, quinolyl, phthaloyl, naphthyridinyl, quinazolinyl, cinnolinyl, pteridinyl, 4 aH-carbazolyl, beta-carbolinyl, phenanthridinyl, acridinyl, naphthyridinyl (perimidinyl), phenanthrolinyl, phenazinyl, isothiazolyl, phenothiazinyl, isothioazinylAzolyl, furazanyl and thiophenesAn oxazine group.

The term "aralkyl" or "arylalkyl" as used herein by itself or as part of another group refers to C discussed above having one aryl substituent_1-6Alkyl radicals, such as benzyl, phenylethyl or 2-naphthylmethyl.

Another aspect of the invention relates to a process for the preparation of formula I, II, III, IV or V.

In embodiments of formulae III, IV or V, the group R^PAre both hydrogen or may be any of a variety of groups useful for protecting sulfur including methoxymethyl, methoxyethoxymethyl, p-methoxybenzyl or benzyl. Sulfur protecting Groups are described in detail in Greene, T.W. and Wuts, P.G.M., Protective Groups in Organic Synthesis, 2 nd edition, John Wiley and Sons, Inc., New York (1991). Protecting group R^PSodium in trifluoroacetic acid, mercuric chloride or liquid ammonia can be removed by suitable methods well known in the art of organic chemistry. In the case of a Lewis acid labile group comprising acetamidomethyl and benzoylaminomethyl groups, R^PCan remain intact. Labeling the ligand with technetium in this case will cleave the protecting group, making the protected diamine dithiol correspond to the unprotected form.

The Tc-99m complex can be prepared as follows. A small amount of non-radiolabelled compound (1-2mg) was dissolved in 100. mu.L EtOH and mixed with 200. mu.L HCl (1N) and 1mL stannous glucoheptonate solution (containing 8-32. mu.g SnCl)₂And 80-320. mu.g sodium glucoheptonate, pH6.67) and 50. mu.L EDTA solution (0.1N). Then add [2 ]^99mTc]Pertechnetate (100-. The reaction was heated at 100 ℃ for 30 minutes and then cooled to room temperature. In TLC (EtOH: concentrated NH)₃9: 1) to check the formation and purity of the product. The mixture was neutralized to ph5.0 with phosphate buffer.

The invention also relates to a method for preparing technetium-99 m complexes, according to which technetium-99 m is reacted in pertechnetate with a suitable Ch-containing compound in the presence of a reducing agent and optionally a suitable chelating agent.

The reducing agent is used to reduce Tc-99m pertechnetate (which is eluted from a molybdenum-technetium generator in physiological saline solution). Suitable reducing agents are, for example, dithionite, formamidinesulfinic acid, diaminoethane disulfonate or suitable metal reducing agents such as Sn (II), Fe (II), Cu (I), Ti (III) or Sb (III). Sn (II) has proven particularly suitable.

For the above-described complex-forming reaction, technetium-99 m is reacted with a suitable compound of the invention in the form of a salt or technetium bound to a relatively weak chelator. In the latter case the desired technetium-99 m complex is formed by ligand exchange. Examples of suitable chelating agents for radionuclide are dicarboxylic acids, such as oxalic acid, malonic acid, succinic acid, maleic acid, phthalic acid, malic acid, lactic acid, tartaric acid, citric acid, ascorbic acid, salicylic acid or derivatives of these acids; phosphorus compounds such as pyrophosphates; or an enolate salt. Citric acid, tartaric acid, ascorbic acid, glucoheptonic acid or derivatives thereof are particularly suitable chelators for this purpose, since the chelate of technetium-99 m with one of these chelators is particularly susceptible to the required ligand exchange.

The most commonly used preparation [ Tc^VO]⁺³N₂S₂The step of complexing is based on^99mTc]Reduction of pertechnetate (a common starting material) from stannous (II) chloride. The labeling step is usually dependent on Tc-99m (Sn) -glucoheptonate and the N₂S₂Tc-99m ligand exchange reaction between ligands. The preparation of stannous (II) chloride and maintaining its constant stannous (II) form is critical to the success of the labeling reaction. In order to stabilize stannous ions sensitive to air, a lyophilization kit is commonly used in nuclear medicine, in which stannous ions are mixed in lyophilized powder form with an excess of glucoheptonate in an inert gas such as nitrogen or argon. The preparation of the lyophilized stannous chloride/sodium glucoheptonate kit ensures that the labeling reaction is reproducible and predictable. These N₂S₂The ligands are generally sensitive to air (thiols are readily oxidized by air) and there is a series of reactions that lead to the decomposition of these ligands. The most convenient and predictable way to preserve these ligands is to produce a lyophilised kit containing 100-500. mu.g of these ligands under argon or nitrogen.

The invention also relates to a process for the preparation of the above formula I, II, III, IV or V. These compounds of the present invention can be prepared by the reactions described in schemes 1-9.

Schemes 1-5 describe a synthetic route to stilbene derivatives of formula I using Wittig reagents.

Route 1

Route 2

Route 3

Route 4

Route 5

Scheme 6 describes a synthetic route to form derivatives of formula II.

Route 6

Scheme 7 describes a synthetic route to form derivatives of formula III.

Route 7

Scheme 8 describes a synthetic route to derivatives of formula IV.

Route 8

Scheme 9 describes a synthetic route to form derivatives of formula IV.

Route 9

Schemes 10 and 11 describe synthetic routes to derivatives of formula I.

Route 10

Route 11

Scheme 12 depicts one synthetic route to the intermediate of formula V.

Route 12

Scheme 13 describes a synthetic route to form derivatives of formula V.

Route 13

When the compounds of the present invention are used as imaging agents, they must be labeled with a suitable emissive halogen isotope. Although it is not limited to¹²⁵I-isotopes are useful for laboratory assays, but they are generally not useful for practical diagnostic purposes, since¹²⁵I has a relatively long half-life (60 days) and low gamma-irradiation (30-65 Kev). Isotope of carbon monoxide¹²³I has a half-life of 13 hours and a gamma energy of 159KeV, so it is expected that the labeling of ligands for diagnostic purposes will be with this isotope. Other isotopes that may be used include¹³¹I (half-life 2 hours). Suitable bromine isotopes include⁷⁷Br and⁷⁶Br。

the emissive halogenated compounds of the present invention lend themselves to easy formation from materials that can be provided to a user in a kit. Kits for forming the imaging agent may contain, for example, vials of physiologically suitable solutions containing the intermediate of formula I, II, III, IV or V, at concentrations and pH suitable for optimal complexing conditions. The user will add an appropriate amount of the radioisotope, e.g., Na, to the vial¹²³I, and an oxidizing agent, such as hydrogen peroxide. The resulting labeled ligand can then be administered intravenously to a patient and receptors in the brain imaged by measuring gamma rays or photo irradiation therefrom.

Since the emissive pharmaceutical composition of the present invention can be easily and simply prepared, the preparation thereof can be easily performed by a user. Accordingly, the present invention relates to a kit comprising:

(1) the non-emittance labelled compounds of the invention, optionally in dry form; optionally adding a pharmaceutically acceptable inert carrier and/or auxiliary substance; and

(2) a reducing agent and optionally a chelating agent;

wherein components (1) and (2) may optionally be mixed; and further wherein instructions describing the above-described method of carrying out the reaction of components (1) and (2) with technetium-99 m in the form of a pertechnetate solution may optionally be included.

Examples of suitable reducing agents and chelating agents for use in the above kits are described above. The pertechnetate solution can be obtained by a user from a molybdenum-technetium generator. Such generators are available in many facilities that perform the steps of the diagnostic activity. As mentioned above, the components (1) and (2) may be mixed, provided that they are compatible. Such a single-component kit, in which the mixed components are preferably lyophilized, is particularly suitable for reaction with the pertechnetate solution in a simple manner by the user.

The emissive diagnostic agent may contain any additives such as a pH control agent (e.g., acid, base, buffer), a stabilizer (e.g., ascorbic acid), or an isotonic agent (e.g., sodium chloride), if desired.

The term "pharmaceutically acceptable salt" as used herein refers to those carboxylic acid salts or acid addition salts of the compounds of the present invention which are, within the scope of sound medical diagnosis, suitable for use in contact with the tissues of patients without undue toxicity, radiation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for the intended use, if possible, also referring to the zwitterionic forms of the compounds of the present invention. The term "salt" refers to the relatively non-toxic, inorganic and organic acid addition salts of the compounds of the present invention. Also included are those salts obtained from non-toxic organic acids such as aliphatic mono-and dicarboxylic acids, e.g., acetic acid, phenyl substituted alkanoic, hydroxyalkanoic and alkanedioic acids, aromatic acids, and aliphatic and aromatic sulfonic acids. These salts may be prepared in situ during the final isolation and purification of these compounds or by separately reacting the purified compound in its free base form with a suitable organic or inorganic acid and isolating the salt thus formed. Other representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthenate, methanesulfonate, glucoheptonate, lactobionate and laurylsulfonate, propionate, pivalate, cyclamate, isethionate and the like. They may include cations based on alkali and alkaline earth metals, such as sodium, lithium, potassium, calcium, magnesium, and the like, as well as non-toxic ammonium, quaternary ammonium, and amine cations, including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. (see, e.g., Berge S.M. et al, Pharmaceutical Salts, J: pharm. Sci.66: 1-19(1977), incorporated herein by reference.)

In the first step of the present imaging method, a labeled compound of formula I, II, III, IV or V is introduced into a tissue or patient in a detectable amount. The compounds are typically part of a pharmaceutical composition and are administered to a tissue or patient by methods well known to those skilled in the art.

For example, the compounds may be administered orally, rectally, parenterally (intravenously, by intramuscular or subcutaneous injection), intracisternally, intravaginally, intraperitoneally, intravesically, topically (powders, ointments or droplets), or as a buccal or nasal spray.

In a preferred embodiment of the invention, the labeled compound is introduced into the patient in a detectable amount and, after a sufficient time for the compound to bind to amyloid deposits, the labeled compound is detected non-invasively within the patient. In another embodiment of the invention, a labeled compound of formula I, II, III, IV or V is introduced into a patient, allowed to bind to amyloid deposits for a sufficient time, and a tissue sample is removed from the patient and removed from the patient to detect the labeled compound in the tissue. In a third embodiment of the invention, a tissue sample is removed from the patient and a labeled compound of formula I, II, III, IV or V is added to the tissue sample. After the compound binds to the amyloid deposits for a sufficient time, the compound is detected.

Administration of the labeled compound to a patient may be by conventional or topical routes of administration. For example, the labeled compound may be administered to the patient so that it is released systemically. Alternatively, the labeled compound may be administered to a particular organ or tissue of interest. For example, to diagnose or track the progression of Alzheimer's disease in a patient, it is desirable to localize and quantify amyloid deposits in the brain.

The term "tissue" refers to a portion of a patient's body. Examples of tissues include brain, heart, liver, blood vessels, and arteries. A detectable amount is the amount of labeled compound that must be detected by the detection method of choice. The amount of marker compound introduced into the patient to provide a test can be readily determined by one skilled in the art. For example, an increased amount of the labeled compound can be administered to the patient until the compound is detected by the selected detection method. A label is introduced to the compound to enable detection of the compound.

The term "patient" refers to humans and other animals. One skilled in the art will also be able to determine, with skill, the sufficient time for the compound to bind to the amyloid deposit. The amount of time required can be readily determined by introducing a detectable amount of a labeled compound of formula I, II, III, IV or V into the patient and then detecting the labeled compound at a different time after administration.

The term "binding" refers to a chemical interaction between a labeled compound and an amyloid deposit. Examples of binding include covalent bonds, ionic bonds, hydrophilic-hydrophilic interactions, hydrophobic-hydrophobic interactions, and complexation.

The person skilled in the art is familiar with various ways of detecting the labeled compound. For example, Magnetic Resonance Imaging (MRI), Positron Emission Tomography (PET), or Single Photon Emission Computed Tomography (SPECT) can be used to detect the emittance-labeled compound. The label added to the compound will depend on the desired detection method. For example, if PET is chosen as the detection method, the compound must have a positive ion-emitting atom, e.g.¹¹C or¹⁸F。

The emissive diagnostic agent should have sufficient emissive properties and emissive concentrations to ensure reliable diagnosis. For example, where the emissive metal is technetium-99 m, an amount of about 0.5-5.0ml of 0.1-50mCi may typically be included when administered. The amount of the compound of formula I, II, III, IV or V may be, for example, sufficient to form a stable chelating compound with the emissive metal.

The chelate compound thus formed is sufficiently stable as an emissive diagnostic agent and can therefore be administered as such or stored until use. The emissive diagnostic agent may contain any additive such as a pH control agent (e.g., acid, base, buffer), a stabilizer (e.g., ascorbic acid), or an isotonic agent (e.g., sodium chloride), if desired.

Imaging of amyloid deposits can also be performed quantitatively, so that the amount of amyloid deposits can be determined.

Preferred compounds for imaging include emitting isotopes such as¹²³I、¹²⁵I、¹³¹I、¹⁸F、⁷⁶Br or⁷⁷Br。

The invention also relates to a method for imaging amyloid deposits. One key condition of in vivo imaging agents of the brain is the ability to cross the entire blood-brain barrier after a bolus injection.

Another aspect of the invention is a method of inhibiting amyloid plaque aggregates. The invention also provides a method of inhibiting amyloid aggregation to form amyloid deposits comprising administering to a patient an amyloid inhibiting amount of a compound of formula I, II, III, IV or V above.

One skilled in the art can readily determine the amount of amyloid inhibition by simply administering a compound of formula I, II, III, IV or V to a patient in increasing amounts until the growth of amyloid deposits is reduced or halted. The growth rate is assessed using imaging as described above or by removing a tissue sample from the patient and observing amyloid deposits therein. The compounds of the present invention may be administered to a patient at dosage levels of from about 0.1 to about 1,000mg per day. For normal adults, a dosage of about 0.01 to about 100mg/kg body weight/day is sufficient for an adult human weighing about 70 kg. However, the specific dosage employed may vary. For example, the dosage may depend on a number of factors including the requirements of the patient, the severity of the condition being treated, and the pharmacological activity of the compound being used. Determination of the optimal dosage for a particular patient is well known to those skilled in the art.

The following examples describe the methods and compositions of the present invention, but are not limited thereto. Other suitable modifications and variations to these conditions and parameters, which are commonly encountered and which are obvious to those skilled in the art, are within the spirit and scope of the present invention.

Example 1

2-iodobenzylphosphonic acid diethyl ester (11)

A mixture of 2-iodobenzyl bromide 10(5g, 16.84mmol) and triethyl phosphite (3.3g, 20mmol) was stirred at 160 ℃. After 4 hours, the mixture was cooled to room temperature the residue was subjected to flash chromatography (EtOAc: Hex, l: 4) to yield 2.3g of 11 (39%).¹H NMR(200MHz，CDCl₃)：δ1.24(t，J＝7.04Hz，6H)，3.40(d，J＝22.00Hz，2H)，4.03(m，4H)，6.91(m，1H)，7.32(m，1H)，7.44(m，1H)，7.82(m，1H)；¹³C NMR(50MHz，CDCl₃)：δ16.27(J＝6.00Hz)，38.31(J＝137.50Hz)，62.16(J＝6.70Hz)，101.16(J＝9.45Hz)，128.23(J＝3.35Hz)，128.45(J＝3.55Hz)，130.60(J＝5.10Hz)，135.36(J＝8.80Hz)，139.60(J＝2.85Hz)。

Example 2

(E) -2' -iodo-N, N-dimethyl-4-stilbenic amine (4)

To a mixture of NaH (2mmol, 80% oil suspension), 3-iodobenzyl phosphonate 2(500mg, 1.42mmol) in 6mL of THF at 80 deg.C under nitrogen was added 4- (dimethylamine) benzaldehyde (210mg, 1.41mmol) dropwise. After overnight at room temperature, NH was added₄Cl solution (saturated, 5mL) and the mixture was taken up with CH₂Cl₂Extraction (3X 30 mL). The combined organic extracts were washed with Na₂SO₄Dried and evaporated to give (E) -2' -iodo-N, N-dimethyl-4-stilbenamine 11 which was purified by flash chromatography (EtOAc: Hex, 1: 9) to give 3(330mg, 67%).¹H NMR(200MHz，CDCl₃)：δ3.06(s，6H)，6.82(m，2H)，6.93-7.02(m，1H)，7.01(d，J＝15.98Hz，1H)，7.25(d，J＝15.99Hz，1H)，7.40(m，1H)，7.53-7.59(m，2H)，7.69(dd，J＝7.88Hz，J＝1.54Hz，1H)，7.95(dd，J＝7.92Hz，J＝1.20Hz，1H)；¹³C NMR(50MHz，CDCl₃): δ 40.26, 100.20, 112.22, 125.12, 125.61, 127.83, 127.87, 127.97, 128.2, 131.66, 139.42, 140.84, 150.23; HRMS: m/z is calculated as C₁₆H₁₆IN: 349.0328, respectively; actually measuring: 349.0342.

example 3

3-iodobenzylphosphonic acid diethyl ester (13)

A mixture of 3-iodobenzyl bromide 12(5g, 16.84mmol) and triethyl phosphite (3.3g, 20mmol) was stirred at 160 ℃. After 4 hours, the mixture was cooled to room temperature. The residue was subjected to flash chromatography (EtOAc: Hex, 1: 4) to give 5.4g of 13 (91%).¹H NMR(200MHz，CDCl₃)：δ1.15(t，J＝7.05Hz，6H)，2.97(d，J＝21.65Hz，2H)，3.92(m，4H)，6.93(t，J＝7.76Hz，1H)，7.17(m，1H)，7.52(m，2H)；¹³C NMR(50MHz，CDCl₃)：δ16.25(J＝5.95Hz)，33.15(J＝137.60Hz)，62.19(J＝6.70Hz)，94.13(J＝3.50Hz)，128.89(J＝6.35Hz)，130.07(J3.00Hz)，133.95(J＝9.10Hz)，135.87(J＝3.55Hz)，138.51(J＝6.65Hz)。

Example 4

(E) -3' -iodo-N, N-dimethyl-4-stilbenic amine (5)

To a mixture of NaH (2mmol, 80% oil suspension) and 3-iodobenzylphosphonate 13(370mg, 1.05mmol) in 5mL THF at 80 deg.C under nitrogen was added 4- (dimethylamine) benzaldehyde (155mg, 1.05mmol) dropwise. After overnight at room temperature, NH was added₄Cl solution (saturated, 5mL), mixture with CH₂Cl₂Extraction (3X 20 mL). The combined organic extracts were washed with Na₂SO₄Dried and evaporated to give (E) -3' -iodo-N, N-dimethyl-4-stilbenamine 5 which was purified by flash chromatography (EtOAc: Hex, 1: 9) to give 3(209mg, 57%).¹HNMR(200MHz，CDCl₃)：δ2.99(s，6H)，6.71(m，2H)，6.77(d，J＝16.41Hz，1H)，7.02(d，J＝16.22Hz，1H)，7.04(t，J＝7.8Hz，1H)，7.36-7.52(m，4H)，7.82(s，1H)；¹³C NMR(50MHz，CDCl₃): δ 40.37, 94.78, 112.38, 112.53, 122.21, 127.76, 128.56, 130.19, 134.76, 135.34, 140.56, 150.36; HRMS: m/z is calculated as C₁₆H₁₆IN: 349.0328, respectively; actually measuring: 349.0302.

example 5

4-Iodobenzylphosphonic acid diethyl ester (15)

A mixture of 4-iodobenzyl bromide 14(5.2g, 17.51mmol) and triethyl phosphite (3.3g, 20mmol) was stirred at 160 ℃. After 4 hours, the mixture was cooled to room temperature. The residue was subjected to flash chromatography (EtOAc: Hex, 1: 4) to yield 3.27g of 15 (53%).¹H NMR(200MHz，CDCl₃)：δ1.24(t，J＝7.04Hz，6H)，3.07(d，J＝21.72Hz，2H)，4.01(m，4H)，7.04(m，2H)，7.62(m，2H)；¹³C NMR(50MHz，CDCl₃)：δ16.24(J＝5.90Hz)，33.21(J＝137.55Hz)，62.04(J＝6.70Hz)，92.15(J＝4.80Hz)，131.31(J＝9.10Hz)，131.57(J＝6.55Hz)，137.43(J＝2.95Hz)。

Example 6

(E) -4' -iodo-N, N-dimethyl-4-stilbenic amine (6)

To a mixture of NaH (2mmol, 80% oil suspension) and 4-iodobenzylphosphonate 15(420mg, 1.19mmol) in 5mL of THF at 80 deg.C under nitrogen was added 4- (dimethylamine) benzaldehyde (180mg, 1.20mmol) dropwise. After overnight at room temperature, water (5mL) was added. The solid formed is filtered off and washed with ether to give crude product 6, which is obtained by reaction with CH₂Cl₂Recrystallization from hexanes purified to give pure 6(156mg, 38%).¹H NMR(200MHz，CDCl₃)：δ2.99(s，6H)，6.71(d，J＝8.60Hz，2H)，6.81(d，J＝16.65Hz，1H)，7.04(d，J＝16.12Hz，1H)，7.21(d，J＝8.15Hz，1H)，7.38(d，J＝8.59Hz，2H)，7.63(d，J＝8.28Hz，2H)；¹³C NMR (50MHz, CDCl 3): δ 40.39, 91.32, 112.38, 123.04, 127.69, 127.73, 128.23, 129.65, 137.55, 137.77, 150.29; HRMS: m/z is calculated as C₁₆H₁₆IN: 349.0328, respectively; actually measuring: 349.0288.

example 7

(E) -4' -iodo-4-O-methoxystilbene alcohol (8)

To a mixture of NaH (2mmol, 80% oil suspension) and 3-iodobenzylphosphonate 13(450mg, 1.27mmol) in 7mL of THF at 80 deg.C under nitrogen was added dropwise anisaldehyde (172mg, 1.27 mmol). After 3 days at room temperature, NH was added₄Cl solution (saturated, 5mL), the mixture was taken up with CH₂Cl₂Extraction (3X 30 mL). The combined organic extracts were washed with Na₂SO₄Dried, evaporated and purified by flash chromatography (EtOAc: Hex, 1: 9) to give (E) -1-iodo-3- [2- (4-methoxyphenyl) vinyl]Benzene 8(400mg, 90%).¹H NMR(200MHz，CDCl₃)：δ3.84(s，3H)，6.84(d，J＝16.29Hz，1H)，6.90(m，2H)，7.05(d，J＝16.30Hz，1H)，7.07(t，J＝7.8Hz，1H)，7.42-7.56(m，4H)，7.85(s，1H)；¹³C NMR(50MHz，CDCl₃): δ 55.32, 94.76, 114.20, 124.85, 125.48, 127.88, 129.58, 129.62, 130.25, 135.00, 135.91, 139.97, 159.62; HRMS: m/z is calculated as C₁₅H₁₃IO: 336.0011, respectively; actually measuring: 336.0006.

example 8

(E) -3' -iodo-4-stilbenol (9)

To 8(350mg, 1.00mmol) CH in a dry ice-acetone bath at-78 deg.C₂Cl₂(200mL) of the solution was added dropwise with BBr₃(10mL, 1M in hexanes). The mixture was warmed to room temperature. Water was added while cooling the reaction mixture at 0 ℃ in an ice bath. MixingCH for things₂Cl₂And (4) extracting. The organic phase was dried and filtered. The filtrate was purified by flash chromatography (EtOAc: Hex, 1: 9) to give 9(296mg, 92%).¹H NMR(200MHz，CDCl₃)：δ4.81(s，1H)，6.83(d，J＝16.17Hz，1H)，6.84(m，2H)，7.03(d，J＝16.32Hz，1H)，7.06(t，J＝7.8Hz，1H)，7.36-7.57(m，4H)，7.84(s，1H)；¹³C NMR(50MHz，CDCl₃): δ 94.75, 115.67, 124.96, 125.49, 128.09, 129.48, 129.87, 130.25, 135.01, 135.96, 139.90, 155.53; HRMS: m/z is calculated as C₁₄H₁₁IO: 321.9855, respectively; actually measuring: 321.9840.

example 9

4-Fluorobenzylphosphonic acid diethyl ester (17)

A mixture of 4-fluorobenzyl bromide 16(1.89g, 10mmol) and triethyl phosphite (1.66g, 10mmol) was stirred at 170 deg.C for 4 hours. The mixture was cooled to room temperature. The residue was subjected to flash chromatography (EtOAc: Hex, 1: 4) to yield 1.4g of 17 (57%).¹H NMR(200MHz，CDCl₃)：δ1.23(t，J＝7.1Hz，6H)，3.10(d，J＝21.4Hz，2H)，3.92(q，J＝7.1Hz，4H)，7.02(m，2H)，7.25(m，2H)。

Example 10

(E) -4-fluoro-4' -dimethylamino-stilbene (7):

to a mixture of phosphate 17(246mg, 1mmol) and 4-dimethylaminobenzaldehyde (149mg, 1mmol) in DMF (2mL) was added KOtBu (224mg, 2mmol) as a solid in portions at room temperature. The resulting mixture was stirred at room temperature overnight. Water (10mL) was added and the solid was collected by suction filtration and washed with water and dried to give 190mg of product (80%).

¹H NMR(200MHz，CDCl₃)：δ2.99(s，6H)，6.71(d，J＝8.9Hz，2H)，6.85(d，J＝16.3Hz，1H)，7.01(t，J＝8.7Hz，2H)，7.40(d，J＝9.0Hz，2H)，7.43(m，2H)；¹³C NMR(200MHz，CDCl₃)：δ41.00，113.01，115.78，116.21，123.76，126.18，127.83，127.99，128.05，129.19，134.91，150.72，164.81.

Example 11

(E) -3-tributylstannyl-4' -dimethylamino-stilbene (18)

5(139mg, 0.38 mmol), bis- (tributyltin) (0.4mL) and Pd (Ph) were added at 90 deg.C₃P)₄(30mg) of a mixed solvent (20mL, two)A mixture of alkane: triethylamine, 3: 1) was stirred overnight. The solvent was removed and the residue was purified by PTLC (Hex: EtOAc, 2: 1) to give 35mg of product (18%, non-optimal yield).¹H NMR(200MHz，CDCl₃)：δ0.94(t，J＝7.2Hz，9H)，1.08-1.66(m，18H)，3.01(s，6H)，6.75(m，2H)，6.94(d，J＝16.3Hz，1H)，7.08(d，J＝16.3Hz，1H)，7.25-7.57(m，6H)；¹³C NMR(50MHz，CDCl₃): δ 9.56, 13.67, 27.37, 29.10, 40.45, 112.45, 124.84, 125.44, 125.98, 127.51, 128.01, 128.51, 134.36, 134.89, 137.41, 142.09, 150.06; HRMS: m/z is calculated as C₂₈H₄₄NSn (MH +): 514.2496, respectively; and (3) analysis: 514.2512.

example 12

Preparation of emissive iodine-labeled ligands

Preparation of the desired compound using a tin methyl iodide alkylation reaction with a tributyltin precursor of 5¹²⁵I-labelled compound. Hydrogen peroxide (50. mu.L, 3% w/v) was added to 50. mu.L of the corresponding tributyltin precursor 18 (1. mu.g/. mu.L EtOH), 50. mu.L 1N HCl and [ [ solution ] ], in a sealed vial¹²⁵I]NaI (1-5 mCi). The reaction was carried out at room temperature 10Min and by adding 100. mu.L of saturated NaHSO₃It is terminated. The reaction mixture was extracted with ethyl acetate (3X 1mL) and then neutralized with saturated sodium bicarbonate solution. The combined extracts were evaporated to dryness. The residue was dissolved in 100. mu.L of EtOH and purified by HPLC using a reverse phase column (Waters C-18ubondpad, 3.9X 300mM) using an isocratic solvent 80% acetonitrile-20% buffer, 3, 3-dimethylglutaric acid (5mM, pH7.0) at a flow rate of 0.8 mL/min. The desired fractions containing the product were collected, concentrated, and extracted again with ethyl acetate. The product without added carrier was evaporated to dryness and redissolved in 100% EtOH (L. mu. Ci/. mu.L), which was the final product¹²⁵The specific activity of the I probe was 2,200Ci/mmole, the emission chemical purity was greater than 95%, stored at-20 ℃ for up to 6 weeks, and then used for in vitro binding studies.

Example 13

Binding assays using solutions of aggregated A.beta. (1-40) peptides

Peptide A β (1-40) was purchased from Bachem (King of Prussia, Pa.) in solid form. Peptide aggregation was performed by gently dissolving the peptide (0.5mg/mL) in a buffer (pH7.4) containing 10mM sodium phosphate and 1mM EDTA. The solution was incubated at 37 ℃ for 36-42 hours with gentle and constant shaking. The procedure is followed in a 12X 75mm borosilicate glass tube¹Binding studies were performed. Aggregated fibrils (10-50 nM in the final assay mixture) were added to a mixture containing 50. mu.l of the emissive ligand (0.01-0.5nM in 40% EtOH) and 10% EtOH in a final volume of 1mL for saturation studies. The final concentration of EtOH was 10%. Nonspecific binding was defined in the presence of 2. mu.M thioflavin T. For inhibition studies, 1mL of the reaction mixture contained 40. mu.l of inhibitor (10)^-5-10^-10M in 10% EtOH) and 0.05nM of an emissive tracer in 40% EtOH. The mixture was incubated at room temperature for 3 hours, and bound and free emission activity was separated by vacuum filtration through Whatman GF/B filters using a Brandel M-24R cell harvester at room temperature followed by washing with 2X 3mL of 10% ethanol. Containing bound I-12The 5 ligand filters were counted in a gamma counter (Packard5000) (counting efficiency 70%). Under these assay conditions, the percentage of specific binding moieties is less than 20% of the total emissive activity. The results of the saturation and inhibition experiments were performed using the software EBDA²Performing nonlinear regression analysis to thereby calculate K_dAnd K_iAnd (4) taking values. Value (K)_inM) are the mean values ± SEM of 3 independent experiments, repeated once per experiment. Other K of the Compounds of formula I_iValues are provided in fig. 1 and 2.

In an in vitro binding assay, preformed A β aggregates of synthetic peptide and [ alpha ], [ beta ]¹²⁵I]TZDM as a ligand, these novel stilbenes show very high binding affinity to the TZ site (2-40nM), while the affinity to the SB site is very low (> 1,000 nM). Clearly, stilbenes containing electron donating groups such as dimethylamino-, -OH, or-OMe groups show excellent binding affinity for Α β aggregates. The benzothiazole ring appears to be unnecessary for binding at the TZ binding site of a β aggregates. This information is very important because it reduces the size of the molecule required to bind to the TZ site (molecular weight fractions of TZDM and 1 are 380 and 349, respectively); which itself significantly increases the flexibility of designing new ligands. Iodinated stilbenes, such as 2 and 5, present structural simplicity, suggesting that the minimum requirement for binding to Α β aggregates may be 3: 1) two benzene rings separated by a vinyl group. 2) One aromatic ring contains an electronegative group, a dimethylamino-, -OH or-OMe group. 3) There appears to be volume tolerance for substitution on the second aromatic ring. To further characterize these compounds, emissive iodinated ligand, [2 ]¹²⁵I]2 is in the presence of Na 2¹²⁵I]I and hydrogen peroxide, whereby the product without added carrier is obtained in high yield (> 95% emission chemical purity). The direct binding test showed that¹²⁵I]2, the new assessment of post-mortem AD brain sections suggested that the new ligand labeled a β plaques, as expected.

Example 14

Biodistribution of novel probes in Normal mice

Under ether anesthesia, 0.15mL of saline solution containing a labeling agent (5-10. mu. Ci) was directly injected into the tail vein of ICR mice (2-3 months old, average body weight 20-30 g). Mice were sacrificed by heart resection at various time points after irradiation. The organ of interest was removed and weighed and its emission activity was counted using an automatic gamma counter (Packard 5000). The percent dose for each organ was calculated by comparing the tissue count to an aliquot of the injected material at the appropriate dilution. The total activity of blood and muscle was calculated under the following assumptions: they are 7% and 40% of the total weight, respectively.

After intravenous injection¹²⁵I]2 in vivo biodistribution studies in normal mice show good brain permeability. At 2, 30, 60 and 120 minutes after injection, brain uptake was 0.84, 1.08, 0.91 and 0.54% dose/organ (blood levels were relatively low at all time points, 5.2-3.6% dose/organ), respectively. And Abeta_1-40The aggregate bound emissive ligands are saturable, K_dIs 0.2 nM.

Although the invention has now been described in detail, it will be appreciated by those skilled in the art that the same can be performed within a wide and equivalent range of conditions, formulations and other parameters without affecting the scope of the invention or any embodiment thereof. All patents, patent applications, and publications cited herein are incorporated by reference in their entirety.

Claims (7)

1. A compound of formula II:

or a pharmaceutically acceptable salt thereof,

z is O or S;

in each case, R⁹、R¹⁰And R¹¹Independently is hydrogen;

R¹²is NR¹³R¹⁴Wherein R is¹³And R¹⁴Is hydrogen, fluorine (C)_1-4) Alkyl or C_1-4An alkyl group; and the number of the first and second electrodes,

x' is¹²⁵I、¹²³I、¹³¹I、¹⁸F、¹⁸Fluorine (C)_1-4) Alkyl group, [2 ]¹⁸Fluorine (C)_1-4) Alkyl radical]Amino group, [2 ]¹⁸Fluorine (C)_1-4) Alkyl radical]An alkylamino group,⁷⁶Br、⁷⁷Br or Sn (alkyl)₃。

2. The compound of claim 1, wherein

Z is O.

3. The compound of claim 1, wherein X' is¹²³I。

4. A pharmaceutical composition comprising a compound according to any one of claims 1 to 3, together with a pharmaceutically acceptable excipient or diluent.

5. A diagnostic composition for imaging amyloid deposits comprising an emittantly labeled compound according to any one of claims 1 to 3; and a pharmaceutically acceptable excipient or diluent.

6. Use of a diagnostic composition according to claim 5 in the manufacture of a medicament for imaging amyloid deposits.

7. Use of the pharmaceutical composition of claim 4 in the manufacture of a medicament for inhibiting amyloid plaque aggregation in a mammal.