Title: Method of inhibiting amyloid protein aggregation and imaging amyloid deposits using aminoindane derivatives
Abstract: The present invention provides compounds of Formula (I) and a method of treating Alzheimer's disease using a compound of Formula (I), wherein: R1 and R2 include alkyl and phenylalkyl; R3 is hydrogen or alkyl; and R4 and R5 include alkyl, alkoxy, carboxyl, alkoxycarbonyl, and nitro. Also provided is a method of inhibiting the aggregation of amyloid proteins using a compound of Formula (I) ##STR1##
and a method of imaging amyloid deposits using compounds of Formula (I).
Patent Number: 6,949,575 Issued on 09/27/2005 to Barta, et al.
| Inventors:
|
Barta; Nancy Sue (Brighton, MI);
Bigge; Christopher Franklin (Ann Arbor, MI)
|
| Assignee:
|
Pfizer Inc. (New York, NY)
|
| Appl. No.:
|
275351 |
| Filed:
|
April 25, 2001 |
| PCT Filed:
|
April 25, 2001
|
| PCT NO:
|
PCT/US01/13254
|
| 371 Date:
|
November 4, 2002
|
| 102(e) Date:
|
November 4, 2002
|
| PCT PUB.NO.:
|
WO01/83425 |
| PCT PUB. Date:
|
November 8, 2001 |
| Current U.S. Class: |
514/381; 514/535; 514/564; 514/567; 514/657; 548/253; 548/254; 560/43; 562/454; 562/457; 564/428 |
| Intern'l Class: |
A01N 043/64; A01N 037/12; A01N 037/44; A61K 031/41; A61K 031/24 |
| Field of Search: |
514/381,535,564,567,657
548/253,254
560/43
562/454,457
564/428
|
References Cited [Referenced By]
U.S. Patent Documents
| 4816456 | Mar., 1989 | Summers.
| |
| 5006561 | Apr., 1991 | Arrowsmith et al.
| |
| 5523314 | Jun., 1996 | Bue-Valleskey et al.
| |
| 5658904 | Aug., 1997 | Ono et al.
| |
| 5716975 | Feb., 1998 | Bue-Valleskey et al.
| |
| 6001331 | Dec., 1999 | Caprathe et al.
| |
| Foreign Patent Documents |
| 2335560 | Jan., 1999 | CA.
| |
| 0535496 | Sep., 1992 | EP.
| |
| 0538134 | Oct., 1992 | EP.
| |
Other References
Nakagawa et al, "Caspase-12 mediates endoplasmic reticulum-specific apoptosis
and cytotoxicity by amyloid-beta" Nature, vo 403, pp. 98-103 (Jan. 6, 2000).
Thal, "Trials to slow progression and prevent disease onset" J. Neural Transm.
[Suppl] vol. 59, pp. 243-249 (2000).
Reichman, W. E., "Alzheimer's Disease: Clinical Treatment Options" vol. 6(22)
Sup., pp. S1125-S1138 (Dec. 2000).
Schenk et al, "Immunization with amyloid-beta attenuates Alzheimer-disease-like
pathology in the PDAPP mouse" Nature, vol. 400, pp. 173-177 (Jul. 8, 1999).
Lue et al, "Soluble Amyloid beta Peptide Concentration as a Predictor of Synaptic
Change in Alzheimer's Disease" American Journal of Pathology, vol. 155(3), pp. 853-.
Mclean et al, "Soluble Pool of A-beta Amyloid as a Determinant of Severity of
Neurodegeneration in Alzheimer's Disease" Annals of Neurology, vol. 46(6), pp.
860-866 (Dec. 1999).
Yan et al, "Membrane-anchored aspartyl protease with Alzheimer's disease beta-secretase
activity" Nature, vol. 402, pp. 533 537 (Dec. 2, 1999).
Bort et al, "Comparative metabolism of the nonsteroidal antiiinflammatory drug
aceclofenac, in the rat, monkey and human." Drug metabolism and disposition: biological
fate of chemicals, vol. 24(9), pp. 969-975 (1996).
Al-Dabbagh and Smith, "Species differences in oxidative drug metabolism: some
basic considerations." Archives of toxicology. Supplement. Archiv fur Toxikologie.
supplement, vol. 7, pp. 219-231 (1984).
Thal, L.J. "Trials to slow progression and prevent disease onset" J. Neural Transmission,
Supplementum, vol. 59, pp. 243-249 (2000).
Reichman, W.E. "Alzheimer's Disease: Clinical Treatment Options" Am. J. Managed
Care, vol. 6, pp. S1133-S1138 (Dec. 2000).
Klein, William, "A.beta. toxicity in Alzheimer's disease: globular oligomers
(ADDLs) as new vaccine and drug targets" Neurochemistry International, vol. 41,
pp. 345-352 (2002).
Reichman, W. E. "Current pharmacologic options for patients with Alzheimer's
disease" Annals of General Hospital Psychiatry, vol. 2:1, online article, 14 pages
(Jan. 2003).
PCT International Search Report PCT/US01/13254.
|
Primary Examiner: Wilson; James O.
Assistant Examiner: Tucker; Zachary C.
Attorney, Agent or Firm: Richardson; P. C., Ling; L. B., Jubinsky; J. A.
Parent Case Text
This application is a 371 application of PCT/US01/13254 filed Apr. 25, 2001,
which claims the benefit of priority to U.S. provisional application Ser. No. 60/201,996
filed May 4, 2000.
Claims
1. A compound of the Formula I
##STR28##
wherein:
R
1 and R
2 independently are hydrogen, C
1-C
3
alkyl, C
2-C
8 alkenyl, C
2-C
8 alkynyl,
(CH
2)
nphenyl or (CH
2)
n substituted
phenyl, provided that one of R
1 and R
2 is other than hydrogen;
R
4 and R
5 independently are hydrogen, halo, C
1-C
8
alkyl, C
2-C
8 alkenyl, C
2-C8 alkynyl,
(CH
2)
nphenyl, (CH
2)
n substituted phenyl,
NO
2, CN, CF
3, C
1-C
8 alkoxy, CO
2R
6,
tetrazolyl, NH(C
1-C
8 alkyl), N(C
1-C
8 alkyl)
2,
or SO
2R
6;
R
3 is hydrogen or C
1-C
8 alkyl;
R
6 is hydrogen, C
1-C
8 alkyl, or (CH
2)
nphenyl
or (CH
2)
n substituted phenyl;
n is an integer from 0 to 4 inclusive;
or a pharmaceutically acceptable salt, ester, or amide thereof.
2. A compound of claim 1 having Formula II.
##STR29##
3. A compound of claim 2 wherein R
6 is hydrogen.
4. A compound of claim 1 having Formula III
##STR30##
wherein R
7 is hydrogen, halo, NO
2, CN, C
1-C
8
alkyl, C
1-C
8 alkoxy, CF
3, NH
2, NH(C
1-C
8
alkyl), or N(C
1-C
8 alkyl)
2.
5. A compound selected from the group consisting of
2-[(2-N,N-di-n-pentylamino)-indan-5-yl]amino-5-nitro-benzoic acid;
Methyl 2-[2-(3,4-Dichloro-benzylamino)-indan-5-ylamino]-benzoate;
2-[2-(3,4-Dichlorobenzylamino)-indane-5-ylamino]-benzoic acid;
2-[2-(3,4-Dichlorobenzylamino)-indan-5-ylamine]-5-nitro-benzoic acid;
2-[2-(3,4-Dichlorobenzylamino)-indan-5-ylamino]-5-methoxy-benzoic acid;
2-(2-Dipentylamino-indan-5-yl-amino)-5-methyl-benzoic acid;
4-(2-Dipentylamino-indan-5-yl-amino)-3-nitro-benzoic acid;
Methyl 2-[5-(3,4-dichlorophenylamino)-indan-2-ylamino]-5-nitro-benzoate;
2-[5-(3,4-Dichlorophenylamino)-indan-2-ylamino]-5-nitro-benzoic acid;
2-[2-(4-Fluorobenzylamino)-indan-5-ylamino]-5-nitro-benzoic acid;
2-{2-[bis-(4-Fluorobenzyl)amino]indan-5-ylamino}-5-nitro-benzoic acid; and
2-[2-(n-Pentylamino)-indan-5-ylamino]-5-nitro-benzoic acid.
6. A method of inhibiting the aggregation of amyloid proteins to form amyloid
deposits, the method comprising administering to a patient in need of inhibition
of amyloid protein aggregation an amyloid protein aggregation inhibiting amount
of a compound of claim 1.
7. A method of inhibiting the aggregation of amyloid proteins to form amyloid
deposits, the method comprising administering to a patient in need of inhibition
of amyloid protein aggregation an amyloid protein aggregation inhibiting amount
of a compound of claim 2.
8. A method of inhibiting the aggregation of amyloid proteins to form amyloid
deposits, the method comprising administering to a patient in need of inhibition
of amyloid protein aggregation an amyloid protein aggregation inhibiting amount
of a compound of claim 3.
9. A pharmaceutical composition comprising a compound of claim 1 together with
an excipient, diluent, or carrier therefore.
10. A pharmaceutical composition comprising a compound of claim 2 together with
an excipient, diluent, or carrier therefore.
11. A pharmaceutical composition comprising a compound of claim 3 together with
an excipient, diluent, or carrier therefore.
12. A pharmaceutical composition comprising a compound of claim 4 together with
an excipient, diluent, or carrier therefore.
Description
FIELD OF THE INVENTION
This invention relates to a method of inhibiting amyloid protein aggregation
and imaging amyloid deposits. More particularly, this invention relates to a method
of inhibiting amyloid protein aggregation in order to treat Alzheimer's disease
using aminoindane derivatives.
BACKGROUND OF THE INVENTION
Amyloidosis is a condition characterized by the accumulation of various
insoluble, fibrillar proteins in the tissues of a patient. The fibrillar proteins
that comprise the accumulations or deposits are called amyloid proteins. While
the particular proteins or peptides found in the deposits vary, the presence of
fibrillar morphology and a large amount of β-sheet secondary structure is
common to many types of amyloids. An amyloid deposit is formed by the aggregation
of amyloid proteins, followed by the further combination of aggregates and/or amyloid proteins.
The presence of amyloid deposits has been shown in various diseases, each with
its particular associated protein, such as Mediterranean fever, Muckle-Wells syndrome,
idiopathetic myeloma, amyloid polyneuropathy, amyloid cardiomyopathy, systemic
senile amyloidosis, amyloid polyneuropathy, hereditary cerebral hemorrhage with
amyloidosis, Alzheimer's disease, Down syndrome, Scrapie, Creutzfeldt-Jacob disease,
Kuru, Gerstrnann-Straussler-Scheinker syndrome, medullary carcinoma of the thyroid,
Isolated atrial amyloid, β
2-microglobulin amyloid in dialysis
patients, inclusion body myositis, β
2-amyloid deposits in muscle
wasting disease, Sickle Cell Anemia, Parkinson's disease, and Islets of Langerhans
diabetes type 2 insulinoma.
A simple, noninvasive method for detecting and quantitating amyloid deposits
in
a patient has been eagerly sought. Presently, detection of amyloid deposits involves
histological analysis of biopsy or autopsy materials. Both methods have major drawbacks.
For example, an autopsy can only be used for a postmortem diagnosis.
The direct imaging of amyloid deposits in vivo is difficult, as the deposits
have many of the same physical properties (i.e., density and water content) as
normal tissues. Attempts to image amyloid deposits directly using magnetic resonance
imaging (MRI) and computer-assisted tomography (CAT) have been disappointing and
have detected amyloid deposits only under certain favorable conditions. In addition,
efforts to label amyloid deposits with antibodies, serum amyloid P protein, or
other probe molecules has provided some selectivity on the periphery of tissues,
but has provided for poor imaging of tissue interiors.
Thus, it would be useful to have a noninvasive technique for imaging and quantitating
amyloid deposits in a patient. In addition, it would be useful to have compounds
that inhibit the aggregation of amyloid proteins to form amyloid deposits.
One of the most devastating diseases associated with amyloid deposits is Alzheimer's
disease. Alzheimer's disease is a degenerative brain disorder characterized clinically
by progressive loss of memory, cognition, reasoning, judgement, and emotional stability
that gradually leads to mental deterioration and ultimately death. Because Alzheimer's
disease and related degenerative brain disorders are a major medical issue for
an increasingly aging population, the need for new treatments and methods for diagnosing
the disorders are needed.
Several classes of compounds have been shown to have activity against Alzheimer's
disease. The only two agents currently approved for clinical treatment of Alzheimer's
disease are the acetylcholinesterase inhibitors tacrine and donepezil (see U.S.
Pat. No. 4,816,456). U.S. Pat. Nos. 5,716,975 and 5,523,314 relate to rhodanine
derivatives useful as hypoglycemic agents and for treating Alzheimer's disease.
The present invention provides a group of aminoindanyl analogs that are inhibitors
of amyloid aggregation and are thus useful for treating Alzheimer's disease. The
compounds are also useful as imaging agents because of their ability to selectively
bind to amyloid proteins.
SUMMARY OF THE INVENTION
The present invention provides compounds of the Formula I
##STR2##
wherein:
R
1 and R
2 independently are hydrogen, C
1-C
8
alkyl, C
2-C
8 alkenyl, C
2-C
8 alkynyl,
(CH
2)
n phenyl or (CH
2)
n substituted
phenyl, provided that one of R
1 and R
2 is other than hydrogen;
R
4 and R
5 independently are hydrogen, halo, C
1-C
8
alkyl, C
2-C
8 alkenyl, C
2-C
8 alkynyl,
(CH
2)
n phenyl, (CH
2)
n substituted phenyl,
NO
2, CN, CF
3, C
1-C
8 alkoxy, CO
2R
6,
tetrazolyl, NH(C
1-C
8 alkyl), N(C
1-C
8 alkyl)
2,
or SO
2R
6;
R
3 is hydrogen or C
1-C
8 alkyl;
R
6 is hydrogen, C
1-C
8 alkyl, or (CH
2)
nphenyl
or (CH
2)
n substituted phenyl;
n is an integer from 0 to 4 inclusive;
or a pharmaceutically acceptable salt, ester, amide, or prodrug thereof.
Preferred compounds have Formula I wherein one of R
1 and R
2
is (CH
2)
nphenyl or (CH
2)
n substituted
phenyl, and R
5 is CO
2R
6, tetrazolyl, or SO
2
R
6, and R
6 is hydrogen.
A preferred group of compounds have Formula II
##STR3##
where R
1, R
2, R
4, and R
6 are as
defined above.
Another preferred group of compounds have Formula III
##STR4##
where in R
4, R
5, and R
6 are as defined above,
and R
7 is hydrogen, halo, NO
2, CN, C
1-C
8
alkyl, C
1-C
8 alkoxy, CF
3, NH
2, NH(C
1-C
8alkyl),
or N(C
1-C
8alkyl)
2.
A further embodiment of this invention is a pharmaceutical composition comprising
a compound of Formula I together with a carrier, excipient, or diluent therefor.
Another embodiment of this invention is a method for inhibiting amyloid aggregation
in a mammal comprising administering an effective amount of a compound of Formula
I. A further method provided is a method of treating Alzheimer's disease and central
and/or peripheral amyloidosis syndromes in mammals comprising administering an
effective amount of a compound of Formula I.
Also provided is a method of imaging amyloid deposits, the method comprising
the steps of:
- a. introducing into a patient a detectable quantity of a labeled compound
of Formula I;
- b. allowing sufficient time for the labeled compound to become associated
with amyloid deposits; and
- c. detecting the labeled compound associated with the amyloid deposits.
In a preferred embodiment of the method, the patient has or is suspected to have
Alzheimer's disease.
In another preferred embodiment, the labeled compound is a radiolabeled compound.
In another preferred embodiment, the labeled compound is detected using MRI.
Also provided is a pharmaceutical composition comprising a compound of Formula I.
DETAILED DESCRIPTION OF THE INVENTION
The term "alkyl" means a straight or branched chain hydrocarbon. Representative
examples of alkyl groups are methyl, ethyl, propyl, isopropyl, isobutyl, butyl,
tert-butyl, sec-butyl, pentyl, and hexyl.
Preferred alkyl groups are C
1-C
8 alkyl.
"Alkenyl" means a carbon chain having one or two points of unsaturation
in the form of double bonds. Examples include ethenyl, prop-2-enyl, and hex-2,4-dienyl.
"Alkynyl" means a carbon chain having one or two triple bonds, for example,
2-butynyl, octa-3,5-diynyl, and the like.
The term "alkoxy" means an alkyl group such as C
1-C
8 alkyl
attached to an oxygen atom. Representative examples of alkoxy groups include methoxy,
ethoxy, tert-butoxy, propoxy, and isobutoxy.
The term "halogen" includes chlorine, fluorine, bromine, and iodine.
The foregoing alkyl, alkenyl, alkynyl, and alkoxy groups can be substituted.
The term "substituted" means that one or more hydrogen atoms in a molecule has
been replaced with another atom or group of atoms. For example, substituents include
halogen, —OH, —CF
3, —NO
2, —NH
2,
—NH(C
1-C
8alkyl), —N(C
1-C
8alkyl)
2,
C
1-C
8 alkyl, —OC
1-C
8 alkyl, —CN,
—CF
3, —CO
2H, —CO
2C
1-C
8
alkyl, SO
2H, and SO
2C
1-C
8 alkyl.
The term "substituted phenyl" means a phenyl ring in which from 1 to 4 hydrogen
atoms have been independently replaced with a substituent, preferably one selected
from the list above. Examples of substituted phenyl include 2,6-dichlorophenyl,
2-methoxycarbonylphenyl, 3-cyanophenyl, 2,3,4,5-tetrafluorophenyl, 3-aminophenyl,
2-hydroxyphenyl, and the like.
The symbol "-" means a covalent bond.
The term "pharmaceutically acceptable salt, ester, amide, and prodrug" as used
herein refers to those carboxylate salts, amino acid addition salts, esters, amides,
and prodrugs of the compounds of the present invention which are, within the scope
of sound medical judgement, suitable for use in contact with the tissues of patients
without undue toxicity, irritation, allergic response, and the like, commensurate
with a reasonable benefit/risk ratio, and effective for their intended use, as
well as the zwitterionic forms, where possible, of the compounds of the invention.
The term "salts" refers to the relatively nontoxic, inorganic and organic acid
addition salts of compounds of the present invention. These salts can be prepared
in situ during the final isolation and purification of the 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. Representative salts include
the hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate,
valerate, oleate, palmitate, stearate, laureate, borate, benzoate, lactate, phosphate,
tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate mesylate,
glucoheptonate, lactiobionate and laurylsulphonate salts, and the like. These may
include cations based on the alkali and alkaline earth metals, such as sodium,
lithium, potassium, calcium, magnesium, and the like, as well as, nontoxic ammonium,
quaternary ammonium and amine cations including, but not limited to ammonium, tetramethylammonium,
tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine,
ethylamine, and the like. (See, for example, Berge S. M., et al., Pharmaceutical
Salts,
J. Pharm. Sci., 66:1-19 (1977) which is incorporated herein by reference.)
Examples of pharmaceutically acceptable, nontoxic esters of the compounds
of this invention include C
1-C
8 alkyl esters wherein the
alkyl group is a straight or branched chain. Acceptable esters also include C
5-C
7
cycloalkyl esters as well as arylalkyl esters such as, but not limited to
benzyl. C
1-C
4 alkyl esters are preferred. Esters of the compounds
of the present invention may be prepared according to conventional methods.
Examples of pharmaceutically acceptable, nontoxic amides of the compounds
of this invention include amides derived from ammonia, primary C
1-C
8
alkyl amines, and secondary C
1-C
8 dialkyl amines wherein
the alkyl groups are straight or branched chain. In the case of secondary amines,
the amine may also be in the form of a 5- or 6-membered heterocycle containing
one nitrogen atom. Amides derived from ammonia, C
1-C
3 alkyl
primary amides, and C
1-C
2 dialkyl secondary amides are preferred.
Amides of the compounds of the invention may be prepared according to conventional methods.
The term "prodrug" refers to compounds that are rapidly transformed in vivo to
yield the parent compound of the above formulas, for example, by hydrolysis in
blood. A thorough discussion is provided in T. Higuchi and V. Stella,
Pro-
drugs
as Novel Delivery Systems, Vol. 14 of the A.C.S. Symposium Series, and in
Bioreversible
Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association
and Pergamon Press, 1987, both of which are incorporated herein by reference.
In addition, the compounds of the present invention can 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 the unsolvated forms for the purposes of the present invention.
The compounds of the present invention can exist in different stereoisometric
forms by virtue of the presence of asymmetric centers in the compounds. It is contemplated
that all stereoisometric forms of the compounds, as well as mixture thereof, including
racemic mixtures, form part of this invention.
In the first step of the present method of imaging, a labeled compound of Formula
I is introduced into a tissue or a patient in a detectable quantity. The compound
is typically part of a pharmaceutical composition and is administered to the tissue
or the patient by methods well-known to those skilled in the art.
In the methods of the present invention, a compound can be administered either
orally, rectally, parenterally (intravenous, by intramuscularly or subcutaneously),
intracisternally, intravaginally, intraperitoneally, intravesically, locally (powders,
ointments or drops), or as a buccal or nasal spray.
Compositions suitable for parenteral injection may comprise physiologically
acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or
emulsions, and sterile powders for reconstitution into sterile injectable solutions
or dispersions. Examples of suitable aqueous and nonaqueous carriers, diluents,
solvents, or vehicles include water, ethanol, polyols (propyleneglycol, polyethyleneglycol,
glycerol, and the like), suitable mixtures thereof, vegetable oils (such as olive
oil), and injectable organic esters such as ethyl oleate. Proper fluidity can be
maintained, for example, by the use of a coating such as lecithin, by the maintenance
of the required particle size in the case of dispersions and by the use of surfactants.
These compositions may also contain adjuvants such as preserving, wetting,
emulsifying, and dispensing agents. Prevention of the action of microorganisms
can be ensured by various antibacterial and antifungal agents, for example, parabens,
chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include
isotonic agents, for example sugars, sodium chloride, and the like. Prolonged absorption
of the injectable pharmaceutical form can be brought about by the use of agents
delaying absorption, for example, aluminum monostearate and gelatin.
Solid dosage forms for oral administration include capsules, tablets, pills,
powders, and granules. In such solid dosage forms, the active compound is admixed
with at least one inert customary excipient (or carrier) such as sodium citrate
or dicalcium phosphate or (a) fillers or extenders, as for example, starches, lactose,
sucrose, glucose, mannitol, and silicic acid; (b) binders, as for example, carboxymethylcellulose,
alignates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants,
as for example, glycerol; (d) disintegrating agents, as for example, agar-agar,
calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates
and sodium carbonate; (e) solution retarders, as for example paraffin; (f) absorption
accelerators, as for example, quaternary ammonium compounds; (g) wetting agents,
as for example, cetyl alcohol and glycerol monostearate; (h) adsorbents, as for
example, kaolin and bentonite; and (i) lubricants, as for example, talc, calcium
stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,
or mixtures thereof. In the case of capsules, tablets, and pills, the dosage forms
may also comprise buffering agents.
Solid compositions of a similar type may also be employed as fillers in soft-
and hard-filled gelatin capsules using such excipients as lactose or milk sugar,
as well as high molecular weight polyethyleneglycols, and the like.
Solid dosage forms such as tablets, dragees, capsules, pills, and granules
can be prepared with coatings and shells, such as enteric coatings and others well
known in the art. They may contain opacifying agents, and can also be of such composition
that they release the active compound or compounds in a certain part of the intestinal
tract in a delayed manner. Examples of embedding compositions which can be used
are polymeric substances and waxes. The active compounds can also be in microencapsulated
form, if appropriate, with one or more of the above-mentioned excipients.
Liquid dosage forms for oral administration include pharmaceutically acceptable
emulsions, solutions, suspensions, syrups, and elixirs. In addition to the active
compounds, the liquid dosage forms may contain inert diluents commonly used in
the art, such as water or other solvents, solubilizing agents and emulsifiers,
as for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate,
benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butyleneglycol, dimethylformamide,
oils, in particular, cottonseed oil, groundnut oil, corn germ oil, olive oil, castor
oil, sesame oil, glycerol, tetrahydrofurfuryl alcohol, polyethyleneglycols, and
fatty acid esters of sorbitan or mixtures of these substances, and the like.
Besides such inert diluents, the composition can also include adjuvants,
such as wetting agents, emulsifying and suspending agents, sweetening, flavoring,
and perfuming agents.
Suspensions, in addition to the active compounds, may contain suspending
agents, as for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol
and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite,
agar-agar and tragacanth, or mixtures of these substances, and the like.
Compositions for rectal administrations are preferably suppositories
which can be prepared by mixing the compounds of the present invention with suitable
nonirritating excipients or carriers such as cocoa butter, polyethyleneglycol,
or a suppository wax, which are solid at ordinary temperatures but liquid at body
temperature and therefore, melt in the rectum or vaginal cavity and release the
active component.
Dosage forms for topical administration of a compound of this invention include
ointments, powders, sprays, and inhalants. The active component is admixed under
sterile conditions with a physiologically acceptable carrier and any preservatives,
buffers or propellants as may be required. Ophthalmic formulations, eye ointments,
powders, and solutions are also contemplated as being within the scope of this invention.
In a preferred embodiment of the invention, the labeled compound is introduced
into a patient in a detectable quantity and after sufficient time has passed for
the compound to become associated with amyloid deposits, the labeled compound is
detected noninvasively inside the patient. In another embodiment of the invention,
a labeled compound of Formula I is introduced into a patient, sufficient time is
allowed for the compound to become associated with amyloid deposits, and then a
sample of tissue from the patient is removed and the labeled compound in the tissue
is detected apart from the patient. In a third embodiment of the invention, a tissue
sample is removed from a patient and a labeled compound of Formula I is introduced
into the tissue sample. After a sufficient amount of time for the compound to become
bound to amyloid deposits, the compound is detected.
The administration of the labeled compound to a patient can be by a general or
local administration route. For example, the labeled compound may be administered
to the patient such that it is delivered throughout the body. Alternatively, the
labeled compound can be administered to a specific organ or tissue of interest.
For example, it is desirable to locate and quantitate amyloid deposits in the brain
in order to diagnose or track the progress of Alzheimer's disease in a patient.
The term "tissue" means a part of a patient's body. Examples of tissues include
the brain, heart, liver, blood vessels, and arteries. A detectable quantity is
a quantity of labeled compound necessary to be detected by the detection method
chosen. The amount of a labeled compound to be introduced into a patient in order
to provide for detection can readily be determined by those skilled in the art.
For example, increasing amounts of the labeled compound can be given to a patient
until the compound is detected by the detection method of choice. A label is introduced
into the compounds to provide for detection of the compounds.
The term "patient" means humans and other animals. Those skilled in the art are
also familiar with determining the amount of time sufficient for a compound to
become associated with amyloid deposits. The amount of time necessary can easily
be determined by introducing a detectable amount of a labeled compound of Formula
I into a patient and then detecting the labeled compound at various times after administration.
The term "associated" means a chemical interaction between the labeled compound
and the amyloid deposit. Examples of associations include covalent bonds, ionic
bonds, hydrophilic-hydrophilic interactions, hydrophobic-hydrophobic interactions,
and complexes.
Those skilled in the art are familiar with the various ways to detect labeled
compounds. For example, MRI, positron emission tomography (PET), or single photon
emission computed tomography (SPECT) can be used to detect radiolabeled compounds.
The label that is introduced into the compound will depend on the detection method
desired. For example, if PET is selected as a detection method, the compound must
possess a positron-emitting atom, such as
11C or
18F.
Another example of a suitable label in a compound of Formula I is an atom
such as
13C,
15N, or
19F which can be detected
using MRI which is also sometimes called nuclear magnetic resonance (NMR). In addition,
the labeled compounds of Formula I may also be detected by MRI using paramagnetic
contrast agents.
Another example of detection is electron paramagnetic resonance (EPR). In
this case, EPR probes which are well-known in the art, such as nitroxides, can
be used.
The imaging of amyloid deposits can also be carried out quantitatively so that
the amount of amyloid deposits can be determined.
The present invention also provides a method of inhibiting the aggregation of
amyloid proteins to form amyloid deposits, by administering to a patient in need
of inhibition of the aggregation of amyloid protein an amyloid protein inhibiting
amount of a compound of Formula I. Those skilled in the art are readily able to
determine an amyloid inhibiting amount by simply administering a compound of Formula
I to a patient in increasing amounts until the growth of amyloid deposits is decreased
or stopped. The rate of growth can be assessed using imaging or by taking a tissue
sample from a patient and observing the amyloid deposits therein.
A patient in need of inhibition of the aggregation of amyloid proteins is a patient
having a central or peripheral disease or condition in which amyloid proteins aggregate.
Examples of such diseases and conditions include Mediterranean fever, Muckle-Wells
syndrome, idiopathetic myeloma, amyloid polyneuropathy, amyloid cardiomyopathy,
systemic senile amyloidosis, amyloid polyneuropathy, hereditary cerebral hemorrhage
with amyloidosis, Alzheimer's disease, Down syndrome, Scrapie, Creutzfeldt-Jacob
disease, Kuru, Gerstmann-Straussler-Scheinker syndrome, medullary carcinoma of
the thyroid, Isolated atrial amyloid, β
2-microglobulin amyloid
in dialysis patients, inclusion body myositis, β
2-amyloid deposits
in muscle wasting disease, Sickle Cell Anemia, Parkinson's disease, and Islets
of Langerhans diabetes type 2 insulinoma.
Also provided by the present invention are compounds of Formula I wherein one
or more atom in the compound has been replaced with a radioisotope. The radioisotope
can be any radioisotope. However,
3H,
123I,
1I,
131I,
11C, and
18F are preferred. Those skilled
in the art are familiar with the procedure used to introduce a radioisotope into
a compound. For example, compounds of Formula I are made where a
12C
atom is replaced by a
13C atom.
The compounds of the present invention can be administered to a patient at dosage
levels in the range of about 0.1 to about 1,000 mg per day, which are "effective
amounts" for inhibiting amyloid formation and treating the above mentioned diseases,
especially Alzheimer's disease. For a normal human adult having a body weight of
about 70 kg, a dosage in the range of about 0.01 to about 100 mg per kilogram of
body weight per day is sufficient. The specific dosage used, however, can vary.
For example, the dosage can 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. The determination of optimum dosages for a
particular patient is well-known to those skilled in the art.
The compounds of Formula I can be prepared by any of several processes, utilizing
readily available starting materials and methods well-known in organic chemistry.
Scheme 1 below illustrates a typical method for making starting materials and the
final products of Formula I. The invention compounds are generally prepared by
reacting an amino indane with a phenyl halide such as a phenyl iodide or phenyl bromide.
Scheme 1 starts with a 2-amino-5-nitro-indane, which can be prepared by reacting
2-amino-indane with nitric acid and sulfuric acid, generally in a solvent such
as trifluoroacetic acid.
The 2-amino-5-nitro-indane is readily reduced to the corresponding 2,5-diaminoindane,
for instance by hydrogenation in the presence of a catalyst such as Raney nickel.
The 5-amino group is then readily phenylated by reaction with a R
4 phenyl
compound
##STR5##
bearing a good leaving group L, for example where L is halo such as iodo.
The reaction is carried out under standard palladium-mediated coupling conditions,
such as in an organic solvent (e.g., toluene) and in the presence of a mild base
(e.g., cesium carbonate). The product, a 2-amino-5-phenylaminoindane, is further
alkylated or phenylated at the 2-amino position, again utilizing standard alkylation methods.
Schemes 2, 3, and 4 illustrate the initial alkylation or phenylation of the
2-amino group of a 2-amino-5-nitro-indane, followed by reduction of the nitro group
and phenylation of the 5-amino group. All of these reactions are carried out under
standard conditions, for example in an unreactive organic solvent, in the presence
of a mild base, and generally at an elevated temperature of about 60° C. to
about 150° C. The products are readily isolated by simply removing the reaction
solvent, for example by evaporation under reduced pressure, and they can be purified
if desired by standard methods such as chromatography, crystallization, distillation,
and the like.
##STR6##
##STR7##
##STR8##
##STR9##
The following detailed examples further illustrate the synthesis of typical compounds
having Formula I. The examples are representative only, and are not intended to
limit the invention in any respect. All references, including patents, cited herein
are incorporated by reference.
Preparation 1
2-Amino-5-nitro-indane sulfate
##STR10##
Trifluoroacetic acid (60 mL) was charged into a flask, cooled in
an ice/water bath, and 2-aminoindane hydrochloride (10.08 g) was added cautiously,
followed by H
2SO
4 (6.0 mL) and HNO
3 (3.0 mL).
The ice bath was removed, and the mixture was allowed to warm to room temperature
and stir for 2 hours. The mixture was then placed in an ice water bath for the
slow addition of diethyl ether (300 mL) over 35 minutes. The mixture was stirred
at room temperature overnight, then the solids were filtered and dried in vacuo
to give 15.43 g of 2-amino-5-nitro-indane sulfate.
Preparation 2
2-(4-Fluorobenzyl)amino-5-nitro-indane and
2-[bis-(4-Fluorobenzyl)amino]-5-nitro-indane
##STR11##
2-Amino-5-nitro-indane sulfate (3 g) from Preparation 1 was
dissolved in 10 mL water and 20 mL 2 M NaOH. The mixture was extracted three times
with 40 mL methyl t-butyl ether, and the organic layers were combined and dried
over MgSO
4/Na
2SO
4, filtered, and concentrated
by rotary evaporation. The residual oil was taken up in 30 mL CH
3CN,
to which 4-fluorobenzyl bromide (2.26 g) was added, followed by K
2CO
3.
The mixture was heated at reflux for 18 hours, cooled to room temperature, filtered,
and concentrated. The residual oil was purified by medium pressure liquid column
chromatography on silica gel (MPLC, biotage column, solvent gradient 99:1 to 85:15
(CH
2Cl
2 /1% NH
4OH in MeOH). The desired mono-alkylated
aminoindane was obtained in 35% yield (1.10 g), and the bis-dialkylated aminoindane
was also isolated (1.38 g, 32% yield). MS (APCI) m/z 287 (M
++1) monoalkylated,
MS (APCI) m/z 395 (M
++1) dialkylated.
Preparation 3
General Conditions For Reduction of the Nitro Group
##STR12##
The nitro substituted indanes from Preparation 2 were reacted with hydrogen in
the presence of Raney nickel to give the corresponding 2-alkyl and 2-dialkylamino-5-aminoindanes.
The 2-amino-5-nitro-indane sulfate from Preparation 1 can be similarly hydrogenated
to provide 2,5-diamino-indane.
Preparation 4
2-n-Pentylamino-5-nitro-indane and
2-N,N-di-n-Pentylamino-5-nitro-indane
##STR13##
2-Amino-5-nitro-indane sulfate from Preparation 1 (2 g) was
dissolved in 20 mL water and brought to pH=11 with 2N NaOH. The aqueous solution
was extracted with 3×30 mL methyl t-butyl ether, the organics were combined,
dried over MgSO
4/Na
2SO
4, filtered, and concentrated.
The residual oil was dissolved in 30 mL acetonitrile, potassium carbonate (1 g)
and 1-bromopentane (1.79 mL) were added. The mixture was stirred at reflux for
18 hours, then filtered, concentrated, and purified by column chromatography (MPLC,
silica, gradient 1:1 to 3:1 ethyl acetate in hexanes) to give the mono-alkylated
amine (0.26 g, 14% yield) and the dialkylated amine (0.99 g, 43% yield). MS mono-alkylated
(APCI) m/z 249.1 (M
++1). MS dialkylated (APCI) m/z 319.2 (M
++1).
Preparation 5
2-(3,4-Dichlorobenzyl)amino-5-nitro-indane
##STR14##
Sulfuric acid (10 mL) was charged into a flask and cooled in an ice/water
bath containing 2-amino-5-nitro-indane hydrochloride (1.68 g), followed by HNO
3
(0.39 mL). The cooling bath was removed, and the mixture was stirred at room temperature
for 30 minutes, and then again cooled in an ice/water bath. Sodium hydroxide (25%
soln) was added carefully to pH=11. The solution was then extracted with 4×60
mL diethyl ether. The combined organic layers were filtered through celite, dried
over MgSO
4/Na
2SO
4, filtered, and concentrated.
The residual black oil was dissolved in THF (25 mL), cooled to 0° C. for the
addition of NaH (0.4 g, 60% dispersion in mineral oil). After 5 minutes, dichloro
benzyl bromide (3.0 g) was added, and the reaction was stirred at room temperature
for 16 hours. The reaction mixture was diluted with 20 mL water and 40 mL Et
2O,
the layers were separated, and the organics were washed with 20 mL water. The aqueous
layers were combined and washed with 30 mL Et
2O, and the organic layers
were combined, dried over MgSO
4/Na
2SO
4, filtered
through celite, and concentrated. The black oil was purified by column chromatography
(MPLC, Isco ReadiSep silica column, solvent gradient 80:20 to 50:50 Hexane/EtOAc)
to give 2-(3,4-dichlorobenzyl)amino-5-nitro-indane (1.05 g, 32% yield). MS (APCI)
m/z 336.1 (M
+-1). The nitro indane was reduced by reaction with hydrogen
and Raney nickel to give 2-(3,4-dichlorobenzyl)amino-5-amino-indane.
Preparation 6
2-Amino-5-(3,4-dichlorophenylamino)-indane
##STR15##
2,5-Diamino-indane (0.77 g) (from Preparation 3) was taken up
in toluene (0.25 M), and nitrogen was bubbled through the solution for 5 minutes.
(S)-(-)-2,2′-bis(di-p-tolyl-phosphino)-1,1′-binaphthyl [(S)-tol-BINAP]
(0.176 g) and palladium dibenzylidene acetone (Pd
2(dba)
3)
(0.123 g), Cs
2CO
3 (2.37 g) and 3,4-dichloro-1-iodo benzene
(1.42 g) was added, and the mixture was heated at reflux for 36 hours. The mixture
was then cooled, diluted with diethyl ether, filtered through a plug of celite,
concentrated, and purified by medium pressure liquid chromatography (silica gel
column, eluted with CH
2Cl
2/MeOH gradient 1% MeOH to 20% MeOH).
The fractions containing any trace of the desired product (mass spec) were combined
and concentrated to give 2-amino-5-(3,4-dichlorophenylamino)-indane (0.54 g, 35%
yield). MS (APCI) m/z 293.1 (M
++1).
EXAMPLE 1
2-[(2-N,N-di-n-pentylamino)-indan-5-yl]amino-5-nitro-benzoic Acid
##STR16##
2-N,N-di-n-Pentylamino-5-amino-indane (prepared
as described in Preparation 3) (0.88 g) was dissolved in toluene (0.25 M), and
nitrogen was bubbled through the solution for 5 minutes. (S)-tol-BINAP (0.16 g)
and Pd
2(dba)
2 (0.073 g), Cs
2CO
3 (1.39
g), and methyl 1-bromo-4-nitro benzoate (0.66 g) were added, and the mixture was
heated at reflux for 18 hours. The mixture was then cooled, diluted with diethyl
ether, filtered through a plug of celite, concentrated, and purified by medium
pressure liquid chromatography (silica gel column, eluted with CH
2Cl
2/MeOH
gradient 1% MeOH to 10% MeOH). The fractions containing any trace of the desired
product (mass spec) were combined and concentrated to give methyl 2-[(2-N,N-di-n-pentylamino)-indane-5-yl]amino-5-nitro-benzoate
(1.28 g, 90% yield). MS (APCI) m/z 468.1 (M
++1). The methyl benzoate
(0.5 g) was dissolved in 20 mL 1:1 THF/MeOH; this solution was treated with 4 mL
1 M LiOH and stirred at room temperature for 24 hours. When the starting material
was no longer observed by mass spec, the mixture was concentrated, and the residual
oil was purified by column chromatography (MPLC, silica, gradient 98:2 to 80:20)(CH
2Cl
2/MeOH+1%
NH
4OH). Fractions containing the desired product were combined, concentrated,
and dried in vacuo overnight to give 2-[(2-N,N-di-n-pentylamino)-indan-5-yl]amino-5-nitro-benzoic
acid (0.240 g, 49% yield). MS (APCI) m/z 454.2 (M
++1). CHN for (C
26H
35N
3O
4)
calc: C, 68.85, H, 7.78, N, 9.26; found: C, 69.37, H, 7.49, N, 8.91.
EXAMPLE 2
Methyl 2-[2-(3,4-Dichloro-benzylamino)-indan-5-ylamino]-benzoate
##STR17##
2-(3,4-Dichloro-benzylamino)-5-amino-indane
(1.08 g) was dissolved in toluene (0.25 M), and nitrogen was bubbled through the
solution for 5 minutes. (S)-tol-BINAP (0.12 g) and Pd
2(dba)
2
(0.08 g), Cs
2CO
3 (1.6 g), and methyl 1-bromo benzoate (0.76
g) were added, and the mixture was heated at reflux for 24 hours. The mixture was
then cooled, diluted with diethyl ether, filtered through a plug of celite, concentrated,
and purified by medium pressure liquid chromatography (silica gel column, eluted
with CH
2Cl
2/MeOH gradient 1% MeOH to 10% MeOH). The fractions
containing any trace of the desired product (mass spec) were combined and concentrated
to give methyl 2-[2-(3,4-dichloro-benzylamino)-indan-5-ylamino]-benzoate (0.98
g, 63% yield). MS (APCI) m/z 441.2 (M
++1). CHN for (C
24H
22Cl
2N
2O
2·0.13CH
3OH)
calc: C, 65.05, H, 5.09, N, 6.29; found: C, 64.72, H, 4.96, N, 6.35.
EXAMPLE 3
2-[2-(3,4-Dichlorobenzylamino)-indane-5-ylamino]-benzoic acid
##STR18##
Methyl 2-[2-(3,4-dichloro-benzylamino)-indan-5-ylamino]-benzoate (0.98 g)
was dissolved in 20 mL 1:1 THF/MeOH, and this solution was treated with 9 mL 50%
NaOH and stirred at room temperature for 24 hours. When the starting material was
no longer observed by mass spec, the mixture was concentrated to remove the MeOH,
extracted with diethyl ether, and the precipitate was collected by filtration and
dried in vacuo overnight to give 2-[2-(3,4-dichlorobenzylamino)-indane-5-ylamino]-benzoic
acid (0.09 g, 10% yield). MS (APCI) m/z 427.1 (M++1). CHN for (C
23H
20Cl
2N
2O
2.1.85HCl)
calc: C, 55.83, H, 4.45, N, 5.66; found: C, 55.45, H, 4.22, N, 5.50.
EXAMPLE 4
2-[2-(3,4-Dichlorobenzylamino)-indan-5-ylamine]-5-nitro-benzoic Acid
By following the procedure of Example 2, 2-(3,4-dichlorobenzylamino)-5-amino-indane
was reacted with methyl 2-bromo-5-nitro benzoate to give methyl 2-[2-(3,4-dichlorobenzyl-amino)-indan-5-yl-amino]-5-nitro-benzoate
(0.65 g, 69% yield). MS (APCI) m/z 486.0 (M
++1).
##STR19##
The methyl benzoate (0.65 g) was dissolved in 10 mL 1:1 THF/MeOH, and this solution
was treated with 2 mL 50% NaOH and 1 mL water and stirred at room temperature for
18 hours. When the starting material was no longer observed by mass spec, the mixture
was concentrated, the solids were dissolved in ethyl acetate/water 1:1. The layers
were separated, and the aqueous layer was extracted with 2×20 mL ethyl acetate.
The organic layers were combined and washed with 10 mL 50% saturated NaCl, dried
over MgSO
4/Na
2SO
4, filtered, and concentrated.
The residual oil was purified by column chromatography (MPLC, silica, gradient
98:2 to 80:20)(CH
2Cl
2/MeOH+1% NH
4OH). Fractions
containing the desired product were combined, concentrated, and dried in vacuo
overnight to give 2-[2-(3,4-dichlorobenzylamino)-indan-5-ylamino]-5-nitro-benzoic
acid (0.38 g, 60% yield). MS (APCI) m/z 470.0 (M
+-1). CHN for (C
23H
19Cl
2N
3O
4·CH
2Cl
2)
calc: C, 55.58, H, 3.94, N, 8.31; found: C, 55.25, H, 3.88, N, 8.32.
EXAMPLE 5
2-[2-(3,4-Dichlorobenzylamino)-indan-5-ylamino]-5-methoxy-benzoic acid
By following the general procedure of Example 2, 2-(3,4-dichlorobenzylamino)-5-amino-indane
was reacted with methyl 2-bromo-5-methoxy benzoate to give methyl 2-[2-(3,4-dichlorobenzylamino)-indan-5-ylamino]-5-methoxy-benzoate
(0.130 g, 11% yield). MS (APCI) m/z 471.0 (M
++1).
##STR20##
The methyl benzoate (0.130 g) was taken up in 20 mL 1:1 THF/MeOH and was treated
with 1 M LIOH (3 mL). The mixture was stirred at room temperature for 48 hours.
The mixture was then concentrated, and the residual oil was purified by chromatography
(MPLC, silica, gradient 95:5 to 75:25 (CH
2Cl
2/MeOH). Fractions
containing the desired product were combined, concentrated, and dried in vacuo
overnight to give 2-[2-(3,4-dichlorobenzylamino)-indan-5-ylamino]-5-methoxy-benzoic
acid (0.03 g, 24% yield). MS (APCI) m/z 457.0 (M
++1). CHN for (C
24H
22Cl
2N
2O
3·0.6H
2O)
calc: C, 61.57, H, 4.98, N, 5.98; found: C, 61.23, H, 4.88, N, 5.69.
EXAMPLE 6
2-(2-Dipentylamino-indan-5-yl-amino)-5-methyl-benzoic acid
##STR21##
2-(2-Dipentylamino)-5-amino-indane (0.5 g) was
taken up in THF (8.7 mL) and cooled to -78° C. for the addition of LiHMDS
(4.0 mL, 1 M in THF). 2-Fluoro-5-methyl benzoic acid (0.27 g) was added after 5
minutes, and the cold bath was removed. The mixture was allowed to warm to room
temperature and stir for 16 hours. The mixture was diluted with diethyl ether and
water. The layers were separated, and the aqueous layer was extracted with 2×20
mL diethyl ether. The organic layers were combined, dried over MgSO
4/Na
2SO
4,
filtered, and concentrated. The residual oil was purified by column chromatography
(MPLC, silica, gradient 95:5 to 75:25)(CH
2Cl
2/MeOH). Fractions
containing the desired product were combined, concentrated, and dried in vacuo
overnight to give 2-(2-dipentylamino-indan-5-yl-amino)-5-methyl-benzoic acid (0.029
g, 4% yield). MS (APCI) m/z 421.2 (M
++1).
EXAMPLE 7
4-(2-Dipentylamino-indan-5-yl-amino)-3-nitro-benzoic Acid
##STR22##
2-(2-Dipentylamino)-5-amino-indane (0.5 g) was
taken up in THF (8.7 mL) and cooled to -78° C. for the addition of LiHMDS
(4.0 mL, 1 M in THF). 4-Fluoro-3-nitro benzoic acid (0.32 g) was added after 5
minutes, and the cold bath was removed. The mixture was allowed to warm to room
temperature and stir for 16 hours. The mixture was diluted with diethyl ether and
water. The layers were separated, and the aqueous layer was extracted with 2×20
mL diethyl ether. The organic layers were combined, dried over MgSO
4/Na
2SO
4,
filtered, and concentrated. The residual oil was purified by column chromatography
(MPLC, silica, gradient 95:5 to 75:25)(CH
2Cl
2/MeOH). Fractions
containing the desired product were combined, concentrated, and dried in vacuo
overnight to give 4-(2-dipentylamino-indan-5-yl-amino)-3-nitro-benzoic acid (0.300
g, 38% yield). MS (APCI) m/z 454.1 (M
++1).
EXAMPLE 8
Methyl 2-[5-(3,4-dichlorophenylamino)-indan-2-ylamino]-5-nitro-benzoate
##STR23##
2-Amino-5-(3,4-dichlorophenylamino)-indane
(0.20 g) was taken up in toluene (0.25 M), and nitrogen was bubbled through the
solution for 5 minutes. (S)-tol-BINAP (0.023 g) and Pd
2(dba)
2
(0.05 g), Cs
2CO
3 (0.31 g), and methyl 2-bromo-4-nitro benzoate
(0.18 g) was added, and the mixture was heated at reflux for 18 hours. The mixture
was then cooled, diluted with diethyl ether, filtered through a plug of celite,
concentrated, and purified by medium pressure liquid chromatography (silica gel
column, eluted with CH
2Cl
2/MeOH gradient 1% MeOH to 20% MeOH).
The fractions containing any trace of the desired product (mass spec) were combined
and concentrated to give methyl 2-[5-(3,4-dichlorophenylamino)-indan-2-ylamino]-5-nitro-benzoate
(0.10 g, 26% yield). MS (APCI) m/z 472.0 (M
++1).
EXAMPLE 9
2-[5-(3,4-Dichlorophenylamino)-indan-2-ylamino]-5-nitro-benzoic acid
##STR24##
By following the procedure of Example 4, methyl 2-[5-(3,4-dichlorophenylamino)-indan-2-ylamino]-5-nitro-benzoate
was reacted with sodium hydroxide in THF/MeOH to give 2-[5-(3,4-dichlorophenylamino)-indan-2-ylamino]-5-nitro-benzoic
acid (0.017 g, 18% yield). MS (APCI) m/z 457.9 (M
++1).
EXAMPLE 10
2-[2-(4-Fluorobenzylamino)-indan-5-ylamino]-5-nitro-benzoic acid
##STR25##
2-(4-Fluorobenzylamino)-5-amino-indane (0.90
g) was taken up in toluene (0.25 M), and nitrogen was bubbled through the solution
for 5 minutes. (S)-tol-BINAP (0.18 g) and Pd
2(dba)
2 (0.08
g), Cs
2CO
3 (1.59 g), and methyl 1-bromo-4-nitro benzoate
(0.75 g) were added, and the mixture was heated at reflux for 48 hours. The mixture
was then cooled, diluted with diethyl ether, filtered through a plug of celite,
concentrated, and partially purified by medium pressure liquid chromatography (silica
gel column, eluted with CH
2Cl
2/MeOH gradient 1% MeOH to 10%
MeOH). The fractions containing any trace of the desired product (mass spec) were
combined and concentrated. The residual crude oil was dissolved in 10 mL 1:1 THF:MeOH
with 1 mL 2N NaOH, and the reaction mixture was stirred at room temperature 4 hours.
When the starting material was no longer observed by mass spec, the mixture was
concentrated, and the residual oil was purified by column chromatography (MPLC,
silica, gradient 95:5 to 75:25 (CH
2Cl
2/1% NH
4OH
in MeOH). Fractions containing the desired product were combined, concentrated,
and dried in vacuo overnight to give 0.83 g of 2-[2-(4-fluorobenzylamino)-indan-5-ylamino]-5-nitro-benzoic
acid. MS (APCI) m/z 422.0 (M
++1). CHN for (C
23H
20F
1N
3O
4·0.68[H
2O])
calc: C, 63.70, H, 4.96, N, 9.69; Found: C, 63.32, H, 4.69, N, 9.65.
EXAMPLE 11
2-{2-[bis-(4-Fluorobenzyl)amino]indan-5-ylamino}-5-nitro-benzoic acid
##STR26##
2[bis-(4-Fluorobenzyl)amino]-5-amino-indane
(1.28 g) was taken up in toluene (0.25 M), and nitrogen was bubbled through the
solution for 5 minutes. (S)-tol-BINAP (0.17 g) and Pd
2(dba)
2
(0.12 g), Cs
2CO
3 (1.6 g), and methyl 1-bromo-4-nitro benzoate
(0.76 g) were added, and the mixture was heated at reflux for 48 hours. The mixture
was then cooled, diluted with diethyl ether, filtered through a plug of celite,
concentrated, and partially purified by medium pressure liquid chromatography (silica
gel column, eluted with CH
2Cl
2/MeOH gradient 1% MeOH to 10%
MeOH). The fractions containing any trace of the desired product (mass spec) were
combined and concentrated to give 1.89 g of the coupled ester. The residual crude
oil was dissolved in 30 mL THF, and 20 mL of this solution was treated with 5 mL
2N NaOH and stirred at room temperature for 12 hours. When the starting material
was no longer observed by mass spec, the mixture was concentrated, and the residual
oil was purified by column chromatography (MPLC, silica, gradient 95:5 to 75:25
(CH
2Cl
2/1% NH
4OH in MeOH). Fractions containing
the desired product were combined, concentrated, and dried in vacuo overnight to
give 0.063 g 2-{2-[bis-(4-fluorobenzyl)amino]indan-5-ylamino}-5-nitro-benzoic acid
pure. MS (APCI) m/z 530.0 (M
++1). CHN for (C
30H
25F
2N
3O
4·CH
3OH)
calc: C, 66.22, H, 5.23, N, 7.46; Found: C, 66.56, H, 5.26, N, 6.97.
EXAMPLE 12
2-[2-(n-Pentylamino)-indan-5-ylamino]-5-nitro-benzoic acid
##STR27##
2-n-Pentylamino-5-amino-indane (0.32 g) was taken
up in toluene (0.2 M), and nitrogen was bubbled through the solution for 5 minutes.
(S)-tol-BINAP (0.076 g) and Pd
2(dba)
2 (0.04 g), Cs
2CO
3
(0.68 g), and methyl 1-bromo-4-nitro benzoate (0.32 g) were added, and the
mixture was heated at reflux for 48 hours. The mixture was then cooled, diluted
with diethyl ether, filtered through a plug of celite, concentrated, and partially
purified by medium pressure liquid chromatography (silica gel column, eluted with
CH
2Cl
2/MeOH gradient 1% MeOH to 10% MeOH). The fractions
containing any trace of the desired product (mass spec) were combined and concentrated
to give 0.42 g of the coupled ester. The residual crude oil was dissolved in 16
mL 1:1 THF/MeOH; this solution was treated with 1 mL 2N NaOH and stirred at room
temperature for 12 hours. When the starting material was no longer observed by
mass spec, the mixture was concentrated, and the residual oil was purified by column
chromatography (MPLC, silica, gradient 98:2 to 80:20 (CH
2Cl
2/MeOH).
Fractions containing the desired product were combined, concentrated, and dried
in vacuo overnight to give 2-[2-(n-pentylamino)-indan-5-ylamino]-5-nitro-benzoic
acid (0.170 g, 30% yield). MS (APCI) m/z 382.0 (M
+-1). CHN for (C
21H
25N
3O
4·0.35CH
2Cl
2)
calc: C, 62.06, H, 6.27, N, 10.17; found: C, 61.67, H, 6.34, N, 10.15.
BIOLOGICAL EXAMPLES
Invention compounds of Formula I can be evaluated in several standard in
vitro and in vivo assays which are well-established as indicative of clinical usefulness
in treating Alzheimer's
