Agents containing malonic acid or derivatives thereof for restoring metal homeostasis

Abstract

The present invention relates to malonic acid and its derivatives according to the following formula (I) for use as an agent for promoting metallothionein biosynthesis in a patient: # imgabs0 # wherein X and Y are each independently selected from the group consisting of O-M +,-OR1 and-NR2R3 wherein M + is selected from the group consisting of a monovalent or multivalent metal cation,-OR1 is selected from the group consisting of a monovalent or multivalent metal cation, and-R2R3 is selected from the group consisting of a monovalent or multivalent metal cation. And R1, R2 and R3 are each independently selected from hydrogen and a saturated or unsaturated, linear, branched or cyclic hydrocarbyl group having 1 to 25 carbon atoms wherein one or more carbon atoms are optionally substituted by O or N, and wherein R2 and R3 are optionally bonded to each other and form a heterocyclic hydrocarbyl group together with a nitrogen atom. The invention also relates to pharmaceutical compositions containing the compounds of formula (I).

Description

[0001] The present invention relates to novel drugs for restoring metal homeostasis, which are potential agents against neurodegenerative diseases. Prior art

[0002] Neurological diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease and prion diseases are age-related neuronal diseases characterized by the accumulation of misfolded proteins and neuronal cell death. All of these diseases are major challenges in current medicine and exist worldwide.

[0003] Alzheimer's disease tops these diseases with almost 50 million cases in 2020. Every 3 seconds, one person on the planet develops dementia, and about 70% of them are diagnosed with Alzheimer's disease. In general, Alzheimer's disease is a type of dementia that affects the patient's thinking, behavior, and memory. The progression of this disease is mild and reduces neurons related to the learning part of the brain. All current methods for treating Alzheimer's disease only provide temporary symptom relief and do not inhibit or reverse the underlying disease mechanism, as these methods were mainly developed based on the amyloid cascade hypothesis, which has not led to satisfactory treatments.

[0004] The second most common neurological disorder of the modern era is Parkinson's disease, a progressive neurological disorder caused by the degeneration of dopamine receptors in the basal ganglia. It is the most common form of Parkinson's disease, a group of disorders characterized by two of the four cardinal signs: bradykinesia, rigidity, tremor, and postural instability. According to the Parkinson's Disease Foundation, approximately 10 million people worldwide suffer from Parkinson's disease. This therefore makes Parkinson's disease the second most common neurodegenerative disease, affecting approximately 1.2% of the world's population over the age of 70. As with Alzheimer's disease, there is still no available therapy that can cure Parkinson's disease. Therefore, the goal of current treatment is to relieve symptoms. More than 40 years after its introduction, levodopa (L-DOPA, L-3,4-dihydroxyphenylalanine) of the following formula remains the most effective therapy for alleviating symptoms associated with Parkinson's disease.

[0005]

[0006] WO 2017 / 142855A1 discloses derivatives of itaconic acid and malonic acid according to the following formulae I and II, respectively:

[0007]

[0008] Where R 1 To R 6Each independently represents hydrogen or an optionally substituted alkyl, alkenyl or alkynyl group with any number of carbon atoms, the optional substituents of which, in addition to hydroxy, may also include a wide range of occasional (hetero)aromatic hydrocarbons. These definitions also include derivatives of succinic acid and maleic acid or fumaric acid, among which itaconic acid (also known as methylenesuccinic acid), malonic acid, its dimethyl ester and fumaric acid and 2-methylfumaric acid (mesaconic acid) are preferably mentioned.

[0009] These compounds are said to act as immunomodulators and therefore can be used to combat a wide range of different types of diseases, including neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease and prion diseases. In preferred embodiments, these compounds are said to inhibit the expression of various proteins. However, in the examples, only free malonic acid and itaconic acid were studied as succinate dehydrogenase inhibitors, which are said to be based on structural similarity to its substrate succinic acid. Potential efficacy for the treatment of neurodegenerative diseases can not be inferred from this.

[0010] In order to overcome this lack of treatment strategies and to find other treatment options, new targets for the treatment of these diseases have been studied. In 2008, Bush and Tanzi proposed in their "Metallhypothese der Alzheimer-Krankheit" (A. Bush and R. E. Tanzi, "Therapeutics for Alzheimer's disease based on the metal hypothesis", Neurotherapeutics 5 (3), 421-432 (2008)) that the imbalance of metal homeostasis is the main cause of neurodegenerative diseases, and therefore drugs that restore metal homeostasis can provide a promising new treatment strategy. Subsequently, metal chelators have been repeatedly proposed as potential agents for the treatment of neurodegenerative diseases, including M. Tosato and A. Di Marco, “Metal Chelation Therapy and Parkinson's Disease: A Critical Review on the Thermodynamics of Complex Formation between Relevant Metal Ions and Promising or Established Drugs”, Biomolecules 9(7), 269 (2019), which lists about 800 substances that have been studied for the treatment of Parkinson's disease in the past few years, among which malonic acid is also mentioned as a metal chelator; and Acevedo et al., “Redox active metals in neurodegenerative diseases”. diseases”, Biol. Inorg. Chem. 24(8), 1141-1157 (2019), which revealed that metal chelators and the like are a means of preventing the misfolding or aggregation of β-amyloid protein (“Aβ”) and α-synuclein, which are the main causes of Alzheimer's disease and Parkinson's disease. In the review by PA Adlard and AIBush (“Metals and Alzheimer's Disease: How Far Have We Come in the Clinic?”, J. Alzheimer's Disease 62(3), 1369-1379 (2018)), a similar hypothesis was made for the chelator clioquinol (5-chloro-7-iodoquinoline-8-ol):

[0011]

[0012] In recent years, the research team of the inventors of the present application has developed a new screening assay using transgenic strains of Caenorhabditis elegans, a nematode from the Rhabditida group, based on this new strategy to restore metal homeostasis, but specifically by upregulating the body's own metallothionein biosynthesis; see Pretsch et al. "Prolongation ofmetallothionein induction combats Aβandα-synuclein toxicity in aged transgenic Caenorhabditis elegans", Sci.Rep.10, 11707 (July 16, 2020). They describe that by extending the endogenous biosynthesis of metallothioneins (i.e., cytoplasmic proteins that bind heavy metals), it is possible to significantly extend the lifespan of the corresponding nematode larvae because this inhibits the expression of toxic beta-amyloid protein ("Aβ") and alpha-synuclein. Emodin (1,3,8-trihydroxy-6-methylanthracene-9,10-dione) has been identified as one of the agents that achieves this:

[0013]

[0014] A subsequent article (D. Pretsch, "Abnormal metal homeostasis as a common drug target to combat neurodegenerative diseases", Neural Regen. Res. 16 (12), 2388-2389 (2021)) further pointed out that not only the effect of emodin, but also the effect of clioquinol cannot be (exclusively) based on its chelating properties, because other effective (especially copper) chelators such as D-penicillamine (2-amino-3-mercapto-3-methylbutyric acid) and diethyl dithiocarbamate

[0015]

[0016] Both failed in tests on mouse models of Parkinson's disease. Also, in this study, D-penicillamine did not show any neuroprotective effect, and the presence of diethyldithiocarbamate even resulted in a higher neurotoxicity. Therefore, the mechanism of action of emodin and clioquinol, which caused the prolongation of the body's own biosynthesis of metallothioneins in the C. elegans test, must not involve binding of heavy metal ions by chelation.

[0017] Hama et al., “Malonic acid suppresses lipopolysaccharide-induced BV2microglia cell activation by inhibiting the p38 MAPK / NF-κB pathway”, Anim. Cells Syst. (Seoul) 25(2), 110-118 (2021) also discloses malonic acid as a potential treatment for Alzheimer's disease and Parkinson's disease, however, due to its ability to inhibit the expression of various cytokines such as interleukins, this can subsequently inhibit various signal transduction pathways associated with the occurrence of inflammation (such as the MAPK pathway).

[0018] In this context, the aim of the present invention was to use a new assay format based on C. elegans as a model organism to identify compounds that are suitable as agents for promoting the biosynthesis of the body's own metallothioneins and thus could be new potential active agents against neurodegenerative diseases. SUMMARY OF THE INVENTION

[0020] The present invention achieves this object by providing malonic acid and its derivatives of the following formula (I), which are used as agents for promoting the biosynthesis of metallothioneins in a patient:

[0021]

[0022] Wherein, X and Y are each independently selected from -O - M + 、-OR 1 and-NR 2 R 3 , where M + is selected from monovalent or polyvalent metal cations, and R 1 , R 2 and R 3 are each independently selected from hydrogen and a saturated or unsaturated, linear, branched or cyclic hydrocarbon group having 1 to 25 carbon atoms, wherein one or more carbon atoms are optionally substituted by O or N, R 2 and R 3 Optionally, they are linked to each other and together with the nitrogen atom form a heterocyclic hydrocarbon group.

[0023] Using the above assay format that they recently developed, the inventors surprisingly found that malonic acid itself and various derivatives of formula (I), i.e., esters, amides and salts thereof, are effective as agents for promoting metallothionein biosynthesis and thereby restoring metal homeostasis in the C. elegans model organism, and are therefore likely to be suitable as agents for combating neurodegenerative diseases. Although unsubstituted malonic acid has been disclosed several times as an agent for treating neurodegenerative diseases due to its chelating properties, it remains unpredictable whether it or any of its derivatives would show a positive effect on the promotion of metallothionein biosynthesis in cells in the nematode assay. This result is particularly surprising for longer chain derivatives, some of which are even substituted with aromatic compounds, and some of which have relatively large structures, which significantly reduces their suitability for forming metal chelates because the central nucleophilic moiety C(=O)-CH 2 The accessibility of the lone electron pair of -C(=O) is restricted.

[0024] However, this efficacy was only demonstrated for malonic acid derivatives in which the malonic acid moiety was unsubstituted at the central α-carbon atom, which is supported by the following Examples and Comparative Examples, which is also surprising and clearly refutes that the reason for this efficacy in this assay is simply the ability of the compound to form a chelate with the metal ion. This is also evidenced by the ineffectiveness of compound (108), N-(3-aminopropyl)-N'-tetrahydroquinolinemalonic acid diamide, in Comparative Example 8, although all other aromatic substituted compounds showed positive results.

[0025] In one embodiment, all compounds thereof are salts of malonic acid, and at least one of X and Y in formula (I) is -O - M + , but preferably, both X and Y are -O - M + , where M + are independently selected from monovalent and multivalent metal cations, and preferably, M + is an alkali metal ion or an alkaline earth metal ion, more preferably an alkali metal ion. Malonates having only one carboxylate ion and malonates in which both carboxyl groups are present as ionized carboxylates have proven to be correspondingly effective. However, in the latter case, the preparation process is somewhat simpler, which is why these are currently preferred according to the present invention.

[0026] In the case of a single malonate (where only one of X and Y represents -OM + and the other represents an ester group -OR 1 or amide group -NR 2 R 3), and in the case of diamides, diesters and esteramides (where both X and Y represent -OR 1 or NR 2 R 3 ), R 1 , R 2 and R 3 Preferably each is independently selected from hydrogen and a saturated or unsaturated, linear, branched or cyclic hydrocarbon radical having 1 to 15 carbon atoms, even more preferably, from hydrogen and a hydrocarbon radical having 1 to 10 carbon atoms, since the lower molecular weight of the compound reduces the amount of active substance to be administered.

[0027] In particular, the compound according to the invention is selected from the following: malonic acid (1), malonic acid diamide (2), disodium malonate (3), dimethyl malonate (4), methyl malonate potassium salt (5), methyl malonate amide (6), N-(3-aminopropyl)malonamide (7), adamantane-1-yl malonate (8), adamantane-1-yl ethyl malonate (9), adamantane-1-yl malonate-N-(3-aminopropyl)amide (10), Adamantane-1-yl malonate-N-(3-aminopropyl)-N-methylamide (11), Adamantane-1-yl malonate-piperazinamide (12), Adamantane-1-yl malonate-4-(4-aminobutyryl)piperazinamide (13), Adamantane-1-yl malonate-4-aminopiperidinamide (14), Adamantane-1-yl malonate-(4-(4-aminobutyryl)amino)piperidinamide (15), Methyl malonate N-(gold N-adamantan-1-yl)malonic acid diamide (16), N-(adamantan-1-yl)malonic acid diamide (17), N-adamantan-1-yl-N'-(3-aminopropyl)malonic acid diamide (18), N-adamantan-1-yl-N'-(3-aminopropyl)-N'-methylmalonic acid diamide (19), N-adamantan-1-yl-N'-piperazinemalonic acid diamide (20), N-adamantan-1-yl-N'-(4-(4-aminobutyryl) piperazine) malonamide (21), N-(3-aminopropyl)-N'-morpholine malonamide (22), N-(3-aminopropyl)-N'-piperidine malonamide (23), N-(3-aminopropyl)-N'-decahydroquinoline malonamide (24), N-(3-aminopropyl)-N'-decahydroisoquinoline malonamide (25), N-(3-aminopropyl)-N'-(6,6-dimethylbicyclo[3.1.1] heptane-2-ylmethyl) malonate diamide (26), malonate adamantane-1-yl ester-N-(3-dimethylaminopropyl)amide (27), malonate adamantane-1-yl ester-N-(3-morpholinylpropyl)amide (28), malonate adamantane-1-yl ester-4-(3-trifluoromethylbenzyl) piperazinamide (29), malonate adamantane-1-yl ester-4-(3-fluorobenzyl) piperazinamide (30), malonate adamantane-1-yl ester-4-(3-trifluoromethylbenzyl) piperazinamide (31), malonate adamantane-1-yl ester-4-(3-fluorobenzyl) piperazinamide (32), malonate adamantane-1-yl ester-4-(3-trifluoromethylbenzyl) piperazinamide (33), malonate adamantane-1-yl ester-4-(3-trifluoromethylbenzyl) piperazinamide (34), malonate adamantane-1-yl ester-4-(3-trifluoromethylbenzyl) piperazinamide (35), malonate adamantane-1-yl ester-4-(3-trifluoromethylbenzyl) piperazinamide (36), malonate adamantane-1-yl ester-4-(3-trifluoromethylbenzyl) piperazinamide (37), malonate adamantane-1-yl ester-4-(3-trifluoromethylbenzyl) piperazinamide (38), malonate adamantane-1-yl ester-4-(3-trifluoromethylbenzyl) piperazinamide (39), malonate adamantane-1-yl ester-4-(3-trifluoromethylbenzyl) piperazinamide (40), malonate adamantane-1-yl ester-4-(3-trifluoromethylbenzyl) piperaz 4-(2-methoxyphenyl)piperazineamide (31), N-(3,5-dimethyl)adamantan-1-ylmalonic acid amide (32), N-(3,5-dimethyladamantan-1-yl)-N'-(3-aminopropyl)malonic acid diamide (33), N-(3,5-dimethyladamantan-1-yl)-N'-(3-morpholinopropyl)malonic acid diamide (34), N-(3,5-dimethyladamantan-1-yl)-N'-(3-morpholinopropyl)malonic acid diamide (35), 1-yl)-N'-piperazine malonamide (35), N-(3,5-dimethyladamantan-1-yl)-N'-4-(3-trifluoromethylbenzyl)piperazine malonamide (36), N-(3,5-dimethyladamantan-1-yl)-N'-4-(3-fluorobenzyl)piperazine malonamide (37), N-(3,5-dimethyladamantan-1-yl)-N'-4-(2-methoxyphenyl)piperazine malonamide amine (38), N-(3,5-dimethyladamantan-1-yl)-N'-4-(4-methoxyphenyl)piperazine malonamide (39), N-(3,5-dimethyladamantan-1-yl)-N'-4-(2,4-dimethoxyphenyl)piperazine malonamide (40), and N-(3,5-dimethyladamantan-1-yl)-N'-4-(2-morpholinyl-2-oxoethyl)piperazine malonamide (41).

[0028] In a preferred embodiment, promotion of metallothionein biosynthesis in a patient is used to restore metal homeostasis and thereby treat a neurodegenerative disease in the patient, which neurodegenerative disease is particularly preferably selected from Alzheimer's disease, Parkinson's disease and Huntington's disease, and since according to the above-mentioned "metal hypothesis", in particular Alzheimer's disease, may be most effectively treatable by restoring metal homeostasis, according to Pretsch et al. (see above), this should be achievable by promoting metallothionein biosynthesis.

[0029] In a second aspect, the present invention accordingly also provides a pharmaceutical composition for therapeutically treating the human or animal body by promoting the biosynthesis of metallothionein in the patient's body, the pharmaceutical composition comprising the compound as defined above for use according to the first aspect and at least one pharmaceutically acceptable excipient, and optionally further comprising one or more other pharmaceutically acceptable ingredients, the composition is preferably also used to treat neurodegenerative diseases, more preferably Alzheimer's disease or Parkinson's disease, in particular Alzheimer's disease. Example

[0030] The invention is described in more detail below with the aid of examples and comparative examples which serve to illustrate the invention and are not to be understood as limiting.

[0031] The compounds of the Examples and Comparative Examples described below are either commercially available or prepared in the manner indicated in the Synthesis Examples and subsequently tested for their suitability as active agents against neurodegenerative diseases using the assay format disclosed in "Pretsch et al." above.

[0032] Test compounds were synthesized using standard reaction sequences, where all starting products were either commercially available or previously synthesized from commercial reagents, which were used in the synthesis without further purification. The spectra were recorded using an Avance AV400 spectrometer from Bruker at 400 MHz. 1 Characterization was performed by H NMR spectroscopy and mass spectra recorded using a LCMS-8040 from Shimadzu.

[0033] In particular, the effectiveness of the following compounds was checked in this first series of tests. Further tests are currently the subject of research work by the inventors.

[0034] So far, the following have been tested as embodiments of the present invention:

[0035]

[0036]

[0037] Example 5: Methyl malonate potassium salt (potassium methylmalonate) (5)

[0038]

[0039] Example 6: Methylmalonate amide (6)

[0040]

[0041] Example 7: N-(3-aminopropyl)malonamide (7)

[0042]

[0043] Example 8: Adamantane-1-yl malonate (adamantyl malonate) (8)

[0044]

[0045] Example 9: Adamantane-1-yl ethyl malonate (adamantyl ethyl malonate) (9)

[0046]

[0047] Example 10: Adamantane-1-yl malonate-N-(3-aminopropyl)amide (10)

[0048]

[0049] Example 11: Adamantane-1-yl malonate-N-(3-aminopropyl)-N-methylamide (11)

[0050]

[0051] Example 12: Adamantane-1-yl malonate piperazinamide (12)

[0052]

[0053] Example 13: Adamantane-1-yl malonate-4-(4-aminobutyryl)piperazinamide (13)

[0054]

[0055] Example 14: Adamantane-1-yl malonate-4-aminopiperidinamide (14)

[0056]

[0057] Example 15: Adamantane-1-yl malonate-(4-(4-aminobutyryl)amino)piperidinamide (15)

[0058]

[0059] Example 16: Methyl malonate N-(adamantan-1-yl)amide (16)

[0060]

[0061] Example 17: N-(adamantan-1-yl)malonic acid amide (17)

[0062]

[0063] Example 18: N-adamantan-1-yl-N'-(3-aminopropyl)malonic acid diamide (18)

[0064]

[0065] Example 19: N-adamantan-1-yl-N'-(3-aminopropyl)-N'-methylmalonic acid diamide (19)

[0066]

[0067] Example 20: N-adamantan-1-yl-N'-piperazinemalonic acid diamide (20)

[0068]

[0069] Example 21: N-adamantan-1-yl-N'-(4-(4-aminobutyryl)piperazine)malonic acid diamide (21)

[0070]

[0071] Example 22: N-(3-aminopropyl)-N'-morpholinemalonic acid diamide (22)

[0072]

[0073] Example 23: N-(3-aminopropyl)-N'-piperidinemalonic acid diamide (23)

[0074]

[0075] Example 24: N-(3-aminopropyl)-N'-decahydroquinolinemalonic acid diamide (24)

[0076]

[0077] Example 25: N-(3-aminopropyl)-N'-decahydroisoquinolinemalonic acid diamide (25)

[0078]

[0079] Example 26: N-(3-aminopropyl)-N'-(6,6-dimethylbicyclo[3.1.1]heptane-2-ylmethyl)malonamide (N-(3-aminopropyl)-N'-(pinan-10-yl)malonamide) (26)

[0080]

[0081] Example 27: Adamantane-1-yl malonate-N-(3-dimethylaminopropyl)amide (27)

[0082]

[0083] Example 28: Adamantane-1-yl malonate-N-(3-morpholinopropyl)amide (28)

[0084]

[0085] Example 29: Adamantane-1-yl malonate-4-(3-trifluoromethylbenzyl)piperazinamide (29)

[0086]

[0087] Example 30: Adamantane-1-yl malonate-4-(3-fluorobenzyl)piperazinamide (30)

[0088]

[0089] Example 31: Adamantane-1-yl malonate-4-(2-methoxyphenyl)piperazineamide (31)

[0090]

[0091] Example 32: Malonic acid-N-(3,5-dimethyl)adamantan-1-ylamide (32)

[0092]

[0093] Example 33: N-(3,5-dimethyladamantan-1-yl)-N'-(3-aminopropyl)malonic acid diamide (33)

[0094]

[0095] Example 34: N-(3,5-dimethyladamantan-1-yl)-N'-(3-morpholinopropyl)malonic acid diamide (34)

[0096]

[0097] Example 35: N-(3,5-dimethyladamantan-1-yl)-N'-piperazinemalonic acid diamide (35)

[0098]

[0099] Example 36: N-(3,5-dimethyladamantan-1-yl)-N'-4-(3-trifluoromethylbenzyl)piperazinemalonic acid diamide (36)

[0100]

[0101] Example 37: N-(3,5-dimethyladamantan-1-yl)-N'-4-(3-fluorobenzyl)piperazinemalonic acid diamide (37)

[0102]

[0103] Example 38: N-(3,5-dimethyladamantan-1-yl)-N'-4-(2-methoxyphenyl)piperazinemalonic acid diamide (38)

[0104]

[0105] Example 39: N-(3,5-dimethyladamantan-1-yl)-N'-4-(4-methoxyphenyl)piperazinemalonic acid diamide (39)

[0106]

[0107] Example 40: N-(3,5-dimethyladamantan-1-yl)-N'-4-(2,4-dimethoxyphenyl)piperazinemalonic acid diamide (40)

[0108]

[0109] Example 41: N-(3,5-dimethyladamantan-1-yl)-N'-4-(2-morpholinyl-2-oxoethyl)piperazinemalonic acid diamide (41)

[0110]

[0111] And as comparative examples, the following compounds were examined:

[0112] Comparative Example 1: Dimethylmalonic acid (101)

[0113]

[0114] Comparative Example 2: Dimethyl methylmalonate (102)

[0115]

[0116] Comparative Example 3: 1-adamantanecarboxylic acid-N-(3-aminopropyl)amide (103)

[0117]

[0118] Comparative Example 4: tert-Butyl 3-aminopropylcarbamate (N-Boc-1,3-propylenediamine) (104)

[0119]

[0120] Comparative Example 5: tert-Butyl 3-(methylamino)propylcarbamate (N-Boc-N'-methyl-1,3-propylenediamine) (105)

[0121]

[0122] Comparative Example 6: 1-adamantanol (106)

[0123]

[0124] Comparative Example 7: 1-Amantadine (107)

[0125]

[0126] Comparative Example 8: N-(3-aminopropyl)-N'-tetrahydroquinolinemalonic acid diamide (108)

[0127]

[0128] Among them, compounds (1) to (7) and (101) to (107) were obtained from commercial sources, and the other compounds (8) to (41) and (108) were prepared as described below.

[0129] Synthesis Example 1

[0130] Preparation of adamantane-1-yl malonate (8)

[0131]

[0132] A solution of 1.0 g (6.57 mmol) of adamantane-1-ol and 0.95 g (6.57 mmol) of 2,2-dimethyl-1,3-dioxane-4,6-dione in toluene was refluxed for 12 h, after which the reaction mixture was allowed to cool to room temperature. Subsequently, 60 ml of saturated NaHCO 3 The aqueous solution was stirred for 2 h, resulting in phase separation. The toluene phase was discarded and the aqueous phase was carefully acidified with 100 ml of 1N HCl. The acidic solution was extracted with dichloromethane and the organic phase was purified by MgSO 4 Drying, filtration and evaporation to dryness afforded 1.5 g of the title compound as colorless crystals.

[0133] 1 H NMR (CD 3OD): δ = 1.71 (m, 6H, adamantyl), 2.10-2.20 (m, 9H, adamantyl), 3.25 (s, 2H), 4.81 (brs, 1H). MS (ESI-): m / z (%) 237 (100, [MH] - ),283(47,[M+HCOO - ]).

[0134] Synthesis Example 2

[0135] Preparation of 1-adamantanylethyl malonate (9)

[0136]

[0137] To a solution of 35 mg of adamantane-1-yl malonate (8) in 10 ml of ethanol was added 2 ml of 4N HCl in dioxane. After stirring at room temperature for 24 h, the reaction mixture was concentrated and the residue was purified by column chromatography (cyclohexane / ethyl acetate 95:5) to give the title compound (35 mg) as a colorless liquid.

[0138] 1 H NMR (CDCl 3 ): δ = 1.25 (t, 3H, J = 7.14Hz), 1.62 (m, 6H), 2.08-2.13 (m, 9H), 3.23 (s, 2H), 4.16 (q, 2H, J = 7.14Hz). MS (ESI+): m / z (%) 267 (13, [M+H] + ),284(6,[M+H 2 O] + ),289(100,[M+Na] + ).

[0139] Synthesis Example 3

[0140] Preparation of malonyl adamantane-1-yl ester-N-(3-aminopropyl)amide (10)

[0141]

[0142] To a solution of 22 mg (0.126 mmol) of tert-butyl 3-aminopropylcarbamate (N-Boc-1,3-propylenediamine) (104), 30 mg (0.126 mmol) of adamantane-1-yl malonate (8) and 45 μl (0.252 mmol) of diisopropylethylamine (DIPEA) in 8 ml of dimethylformamide (DMF) cooled to 0° C. was added 54 mg (0.126 mmol) of (1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylaminomorpholinium hexafluorophosphate (COMU). After stirring for 2 h and allowing to warm to room temperature, the reaction mixture was diluted with ethyl acetate and washed with 1N HCl and saturated NaHCO 3 Wash with aqueous solution and MgSO 4 Dry, filter and evaporate to dryness.The residue (47 mg) containing Boc protected adamantane-1-yl malonate-N-(3-aminopropyl)amide (10a) was used in the next step without further purification.

[0143] MS (ESI+): m / z (%) 395 (100, [M+H] + ),417(100,[M+Na] + ),811(100,[2M+Na] + ).

[0144] The residue thus obtained was dissolved in 8 ml of dichloromethane, 1.26 ml of 4N HCl in dioxane were added and the mixture was stirred at room temperature for 2 hours. The volatiles were then removed in vacuo to give the hydrochloride of the title compound (10) (34 mg) in the form of a yellow oil.

[0145] 1 H NMR (CDCl 3 ): δ=1.60-1.72(m,10H),2.10-2.18(m,9H),2.85(s,2H),3.22-3.25(m,1H) ),3.62-3.70(m,2H),3.75-3.79(m,1H).MS(ESI+):m / z(%)295(100,[M+H] + ),589(80,[2M+H] + ),611(15,[2M+Na] + ).

[0146] Synthesis Example 4

[0147] Preparation of malonyl adamantane-1-yl ester-N-(3-aminopropyl)-N-methylamide (11)

[0148]

[0149] To a solution of adamantyl malonate (8) (289 mg, 1.211 mmol), tert-butyl 3-(methylamino)propylcarbamate (105) (228 mg, 1.211 mmol) and triethylamine (TEA) (340 μl, 2.422 mmol) in DMF (6 ml) was slowly added 1 ml of 1.6 M solution of propanephosphonic anhydride (T3P) in ethyl acetate and stirred at room temperature for 1 h. The reaction mixture was then mixed with saturated brine for hydrolysis and extracted with dichloromethane. The extract was purified by MgSO 4 Dried, filtered and evaporated to dryness.The residue was purified by column chromatography (dichloromethane / ethyl acetate 70:30) to give Boc-protected adamantane-1-yl malonate-N-(3-aminopropyl)-N-methylamide (11a) (370 mg) as a colorless oil.

[0150] MS(ESI+):m / z(%)409(100,[M+H] + ,431(100,[M+Na] + ),309(100,[M-Boc] + ),839(100,[2M+Na] + ).

[0151] The product thus obtained (0.96 mmol) was dissolved in 20 ml of dichloromethane, 9.6 ml of 4N HCl in dioxane were added and the mixture was stirred at room temperature for 1.5 h. The volatile components were then removed in vacuo to give the hydrochloride of the title compound (11) (260 mg) in the form of a yellow oil.

[0152] 1 H NMR (DMSO-d 6 ): δ=1.63-1.75(m,10H),2.10-2.20(m,9H),2.85(s,2H),2.97(s,3H)3.22-3.25 (m,1H),3.61-3.72(m,2H),3.73-3.82(m,1H).MS(ESI+):m / z(%)309(100,[M+H] + ),331(17,[M+Na] + ),617(77,[2M] + ).

[0153] Synthesis Example 5

[0154] Preparation of malonate adamantane-1-yl ester piperazinamide (12)

[0155]

[0156] Analogously to Synthesis Example 4, the product was synthesized from adamantyl malonate (8) (150 mg, 0.630 mmol), tert-butyl 1-piperazinecarbamate (117 mg, 0.630 mmol) and TEA (176 μl, 1.26 mmol) in DMF (7 ml) together with 512 μl of a 1.6 M solution of T3P in ethyl acetate. The residue was purified by column chromatography (dichloromethane / ethyl acetate 90:10→50:50) to give the Boc-protected title compound (12a) (232 mg) in the form of a white solid.

[0157] MS (ESI+): m / z (%) 407 (100, [M+H] + ),429(100,[M+Na] + ),836(100,[2M+Na] + ).

[0158] The product thus obtained (0.571 mmol) was deprotected similarly to Synthesis Example 4 to obtain hydrochloride (169 mg) of the title compound (12) in the form of a white solid.

[0159] 1 H NMR (DMSO-d 6 ): δ=1.6(t,6H,J=3.0Hz),2.03(d,6H,J=3.0Hz),2.11(brs,3H),3.05(dt,4H ,J=22.6,5.2Hz),3.32(brs,1H),3.48(s,2H),3.63(dt,4H,J=22.6,5.2Hz).

[0160] MS(ESI+):m / z(%)307(100,[M+H] + ),329(25,[M+Na] + ),635(15,[2M+Na] + ).

[0161] Synthesis Example 6

[0162] Preparation of malonyl adamantane-1-yl ester-4-(4-aminobutyryl)piperazinamide (13)

[0163]

[0164] Analogously to Synthesis Example 4, the malonate adamantane-1-yl ester piperazinamide (12) (180 mg, 0.525 mmol), 4-(tert-butoxycarbonylamino)butyric acid (107 mg, 0.525 mmol) and triethylamine (TEA) (219 μl, 1.575 mmol) in DMF (7 ml) were synthesized together with 427 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 80:20) gave the Boc-protected title compound (13a) (190 mg) in the form of a white solid.

[0165] MS (ESI+): m / z (%) 492 (100, [M+H] + ),514(100,[M+Na] + ),392(72,{M+H-Boc] + ).

[0166] The product thus obtained (0.386 mmol) was deprotected similarly to Synthesis Example 4 to obtain hydrochloride (151 mg) of the title compound (12) in the form of a white powder.

[0167] 1 H NMR (DMSO-d 6 ):1.59(brs,6H),1.76(quint,2H,J=7.4Hz),2.03(d,6H,J=3Hz),2.10(brs,3H),2.45(quint,2H,J=7.4 Hz),2.74-2.83(m,2H),3.34(brs,2H),3.40-3.45(m,8H),3.54(s,2H).MS(ESI+):m / z(%)392(100,[M+H] + ),414(100,[M+Na] + ),430(10,[M+K] + ),805(69,[2M+Na] + ).

[0168] Synthesis Example 7

[0169] Preparation of malonyl adamantane-1-yl ester-4-aminopiperidinamide (14)

[0170]

[0171] Similar to Synthesis Example 4, by malonate adamantane-1-yl ester (8) (150mg, 0.630mmol), piperidin-4-ylcarbamic acid tert-butyl ester (126mg, 0.630mmol) and triethylamine (TEA) (176μl, 1.26mmol) in DMF (7ml) and 512μl of 1.6M solution of T3P in ethyl acetate, synthesize together. Purification by column chromatography (dichloromethane / ethyl acetate 90:10→80:20) obtains the Boc-protected title compound (14a) (236mg) in the form of a white solid.

[0172] MS(ESI+): m / z(%)421(100,[M+H] + ),443(100,[M+Na] + ),864(100,[2M+Na] + ).

[0173] The product thus obtained (0.561 mmol) was deprotected similarly to Synthesis Example 4 to obtain hydrochloride (175 mg) of the title compound (14) in the form of a white powder.

[0174] MS(ESI+):m / z(%)321(100,[M+H] + ),343(7,[M+Na] + ),641(24,[2M+H] + ),663(95,[2M+Na] + ).

[0175] Synthesis Example 8

[0176] Preparation of malonyl adamantane-1-yl ester-(4-(4-aminobutyryl)amino)piperidinamide (15)

[0177]

[0178] Analogously to Synthesis Example 4, the malonate adamantane-1-yl ester-4-aminopiperidinamide (14) (140 mg, 0.392 mmol), 4-(tert-butoxycarbonylamino)butyric acid (80 mg, 0.392 mmol) and triethylamine (TEA) (164 μl, 1.176 mmol) in DMF (7 ml) were synthesized together with 319 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 80:20→ethyl acetate) gave the Boc-protected title compound (15a) (83 mg) in the form of a yellow oil.

[0179] MS(ESI+):m / z(%)506(100,[M+H] + ),528(100,[M+Na] + ),406(78,[M-Boc+H] + ).

[0180] The product thus obtained (0.164 mmol) was deprotected similarly to Synthesis Example 4 to obtain hydrochloride (80 mg) of the title compound (14) in the form of a yellow powder.

[0181] 1 H NMR (DMSO-d 6 ): δ=1.64(dt,3H,J=13.1,2.8Hz), 1.66(dt,3H,J=13.1,2.8Hz), 1.74(sept,3 H,J=2.8Hz),1.80-1.87(m,6H),2.09-2.13(m,6H),2.16(t,2H,J=7.4Hz),2.6 5(t,2H,J=7.7Hz),3.04-3.13(m,2H),3.35(ddd,2H,J=14.7,6.8,2.8Hz),3.5 5(s,2H),4.11-4.21(m,1H),7.97(brs,2H).MS(ESI+):m / z(%)406(100,[M+H] + ),428(100,[M+Na] + ),811(100,[2M+H] + ),833(53,[2M+Na] + ).

[0182] Synthesis Example 9

[0183] Preparation of methyl malonate N-(adamantan-1-yl)amide (16)

[0184]

[0185] Malonate methyl chloride (322 μl, 3.005 mmol) was dissolved in 30 ml of dichloromethane under an inert gas atmosphere and cooled to -20°C, and then a solution of 1-adamantanamine (107) (1.0 g, 6.612 mmol) in 10 ml of dichloromethane was slowly added using a syringe. The reaction mixture was stirred at room temperature for 1 hour and saturated with NH 4 The organic phase was quenched with aqueous Cl solution and extracted with dichloromethane. 4 Drying, filtration and evaporation to dryness gave the title compound (760 mg) as white crystals.

[0186] 1 H NMR (CDCl 3 ): δ=1.68(m,6H),2.01-2.10(m,9H),3.22(s,2H),3.74(s,3H),6.64(brs,1H).MS(ESI+):m / z(%)252(100,[M+H] + ),274(100,[M+Na] + ),503(78,[2M+H] + ).

[0187] Synthesis Example 10

[0188] Preparation of N-(adamantan-1-yl)malonamide (17)

[0189]

[0190] To 750 mg (2.98 mmol) of methyl malonate N-(adamantan-1-yl)amide (16) was added 30 ml of THF / H 2 O (2:1), 6 ml of 1 M NaOH aqueous solution was added and stirred at room temperature. After 1 h, THF was removed on a rotary evaporator, and the remaining aqueous solution was acidified with 2N HCl and extracted with ethyl acetate. The organic phase was purified by MgSO 4 Drying, filtration and evaporation to dryness gave the title compound (17) (670 mg) as an off-white powder.

[0191] MS (ESI-): m / z (%) 236 (100, [MH] - ),473(100,[2M-H] - ).

[0192] Synthesis Example 11

[0193] Preparation of N-adamantan-1-yl-N'-(3-aminopropyl)malonic acid diamide (18)

[0194]

[0195] To a solution of N-(adamantan-1-yl)malonamide (17) (50 mg, 0.211 mmol), tert-butyl 3-aminopropylcarbamate (104) (37 mg, 0.211 mmol) and 4-dimethylaminopyridine (DMAP) (13 mg, 0.106 mmol) in dichloromethane (8 ml) was added diisopropylcarbodiimide (DIC) (40 μl, 0.253 mmol). After stirring for 4 h while allowing to warm to room temperature, the reaction mixture was evaporated to dryness. The residue (59 mg, 0.202 mmol) containing Boc-protected N-adamantan-1-yl-N'-(3-aminopropyl)malonamide (18a) was used in the next step without further purification.

[0196] MS(ESI+):m / z(%)394(100,[M+H] + ),415(100,[M+Na] + ),294(70,[M-BOC+H] + ),787(40,[2M+H] + ).

[0197] The residue thus obtained was dissolved in 6 ml of dichloromethane, 2 ml of 4N HCl in dioxane were added and the mixture was stirred at room temperature for 2 hours. The volatiles were then removed in vacuo to give the hydrochloride salt of the title compound (18) (55 mg) in the form of a yellow oil.

[0198] 1 H-NMR (DMSO-d 6 ): δ = 1.61 (m, 6H, adamantyl), 1.68 (quint, 2H, J = 7.1 Hz), 1.90-2.00 (m, 9H, adamantyl), 2.74-2.82 (m, 2H), 2.98 (s, 2H), 3.12 (q, 2H, J = 6.42 Hz), 7.58 (s, 1H). MS (ESI+): m / z (%) 294 (90, [M+H] + ),316(100,[M+Na] + ),608(45,[2M+Na] + ).

[0199] Synthesis Example 12

[0200] Preparation of N-adamantan-1-yl-N'-(3-aminopropyl)-N'-methylmalonic acid diamide (19)

[0201]

[0202] Similar to Synthesis Example 4, N-(adamantan-1-yl)malonamide (17) (50 mg, 0.211 mmol), tert-butyl 3-(methylamino)propylcarbamate (105) (40 mg, 0.211 mmol) and triethylamine (TEA) (60 μl, 0.422 mmol) in DMF (10 ml) were synthesized together with 175 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 70:30) gave the Boc-protected title compound (19a) (83 mg) in the form of a yellow oil.

[0203] MS(ESI+):m / z(%)408(100,[M+H] + ),430(100,[M+Na] + ),308(100,[M-BOC+H] + ),815(50,[2M+H] + ).

[0204] The product thus obtained (46 mg, 0.113 mmol) was deprotected similarly to Synthesis Example 4 to give hydrochloride (33 mg) of the title compound (19) in the form of a yellow oil.

[0205] 1 H NMR (DMSO-d 6 ): δ = 1.61 (m, 6H, adamantyl), 1.74 (quint, 2H, J = 5.5 Hz), 1.97-2.07 (m, 9H, adamantyl), 2.75-2.84 (m, 2H), 2.93 (s, 3H), 3.26 (s, 2H), 3.35 (t, 2H, J = 5.5 Hz), 3.41 (brs, 2H), 7.50 (s, 1H). MS (ESI+): m / z (%) 308 (100, [M+H] + ),330(33,[M+Na] + ),615(100,[2M+H] + ).

[0206] Synthesis Example 13

[0207] Preparation of N-adamantan-1-yl-N'-piperazinemalonic acid diamide (20)

[0208]

[0209] The synthesis was carried out similarly to Example 4 by commercial malonic acid-4-(tert-butoxycarbonyl)piperidinamide (250 mg, 0.918 mmol), 1-adamantanamine (107) (139 mg, 0.918 mmol) and triethylamine (TEA) (265 μl, 1.84 mmol) in DMF (17 ml) together with 750 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 90:10→50:50) gave the Boc-protected title compound (20a) (260 mg) in the form of a white solid.

[0210] 1 H NMR (CDCl 3 ): δ=1.46(s,9H),1.67(t,7H,J=3.9Hz),1.98(d,6H,J=3.1Hz),2.06(brs,3H),3.24(s,2H),3.40-3 .46(m,4H),3.52-3.55(m,2H),3.58-3.61(m,2H),6.73(brs,1H).MS(ESI+):m / z(%)406(100,[M+H] + ),428(100,[M+Na] + ),833(2M+Na] + ).

[0211] The product thus obtained (0.647 mmol) was deprotected similarly to Synthesis Example 4 to obtain hydrochloride (192 mg) of the title compound (20) in the form of a white powder.

[0212] 1 H NMR (DMSO-d 6 ):1.61(brs,6H),1.9(d,6H,J=2.95Hz),2.00(brs,3H),3.00(brs,2H),3.10( brs,2H),3.66-3.68(m,4H),4.01(brs,2H).MS(ESI+):m / z(%)306(100,[M+H] + ),329(7,[M+Na] + ),633(95,[2M+Na] + ).

[0213] Synthesis Example 14

[0214] Preparation of N-adamantan-1-yl-N'-(4-(4-aminobutyryl)piperazine)malonic acid diamide (21)

[0215]

[0216] The synthesis was carried out similarly to Synthesis Example 4 from the hydrochloride salt (140 mg, 0.410 mmol) of N-adamantan-1-yl-N'-piperazine malonate diamide (20), 4-(tert-butoxycarbonylamino)butyric acid (83 mg, 0.410 mmol) and triethylamine (TEA) (171 μl, 1.23 mmol) in DMF (7 ml) together with 333 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (ethyl acetate / methanol 95:5) gave the Boc-protected title compound (21a) (180 mg) in the form of a white solid.

[0217] MS(ESI+):m / z(%)491(100,[M+H] + ),513(100,[M+Na] + ),391(38,[M-Boc+H] + ).

[0218] The product thus obtained (0.367 mmol) was deprotected similarly to Synthesis Example 4 to give hydrochloride (150 mg) of the title compound (21) in the form of a white powder.

[0219] 1 H NMR (DMSO-d 6 ):1.59(brs,6H),1.77(quint,2H,J=6.4Hz),1.89(brs,6H),1.98(brs,3H),2.43-2.47(m, 2H),2.76-2.81(m,2H),3.26-3.30(m,2H),3.39-3.45(m,8H),4.34(brs,2H),7.61(s,1H).

[0220] Synthesis Example 15

[0221] Preparation of N-(3-aminopropyl)-N'-morpholinemalonamide (22)

[0222]

[0223] Similar to Synthesis Example 4, commercial N-morpholinylmalonamide (90 mg, 0.520 mmol), tert-butyl 3-aminopropylcarbamate (N-Boc-1,3-propylenediamine) (104) (91 mg, 0.520 mmol) and triethylamine (TEA) (145 μl, 1.04 mmol) in DMF (5 ml) were synthesized together with 423 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (ethyl acetate / methanol 95:5) gave the Boc-protected title compound (22a) (90 mg) in the form of a yellow oil.

[0224] MS(ESI+):m / z(%)330(100,[M+H] + ),352(100,[M+Na] + ),230(100,[M-Boc+H] + ),681(100,[2M+Na] + ).

[0225] The product thus obtained (0.273 mmol) was deprotected similarly to Synthesis Example 4 to give hydrochloride (76 mg) of the title compound (22) in the form of a yellow oil.

[0226] MS(ESI+):m / z(%)230(100,[M+H] + ),252(100,[M+Na] + ),459(11,[2M+H] + ),481(100,[2M+Na] + ).

[0227] Synthesis Example 16

[0228] Preparation of N-(3-aminopropyl)-N'-piperidinylmalonic acid diamide (23)

[0229]

[0230] The synthesis was carried out analogously to the combination of Synthesis Examples 9, 10 and 4, wherein first, analogously to Synthesis Example 9, methyl malonate N-piperidinamide was prepared from methyl malonate chloride (250 μl, 2.33 mmol) and piperidine (506 μl, 5.126 mmol) at -20°C in 10 ml of dichloromethane, and then analogously to Synthesis Example 10 in THF / H 2 O (2:1) with aqueous NaOH to produce malonic acid-N-piperidinamide.

[0231] MS(ESI+):m / z(%)172(100,[M+H]+ ),194(50,[M+H] + ).

[0232] Then, similar to Synthesis Example 4, it (280 mg, 1.636 mmol) was reacted with tert-butyl 3-aminopropylcarbamate (N-Boc-1,3-propylenediamine) (285 mg, 1.636 mmol) and TEA (456 μl, 3.272 mmol) in DMF (6 ml) with 1.33 ml of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 50:50→ethyl acetate) gave the Boc-protected title compound (23a) in the form of a colorless oil (370 mg).

[0233] 1 H NMR (CDCl 3 ): δ=1.43(s,9H),1.53-1.60(m,4H),1.63-1.68(m,5H),3.14(q,2H,J=6.5Hz),3.31(s,2H),3.33(q,2H ,J=6.5Hz),3.45(t,2H,J=4.8Hz),3.56(t,2H,J=4.8Hz),7.78(s,1H).MS(ESI+):m / z(%)328(100,[M+H] + ),350(100,[M+Na] + ),228(80,[M-Boc+H] + ),677(100,[2M+Na] + ).

[0234] The thus obtained product (1.13 mmol) was deprotected similarly to Synthesis Example 4 to obtain hydrochloride (330 mg) of the title compound (23) in the form of a white powder.

[0235] 1 H NMR (DMSO-d 6 ): δ=1.37-1.43(m,2H),1.45-1.50(m,2H),1.53-1.59(m,2H),1.69(quint,2H,J=8.3Hz),2.75-2 .83(m,2H),3.12(q,2H,4.8Hz),3.35-3.41(m,4H),7.96(s,1H).MS(ESI+):m / z(%)228(100,[M+H] + ),250(16,[M+Na] + ),455(40,[2M+H] + ).

[0236] Synthesis Example 17

[0237] Preparation of N-(3-aminopropyl)-N'-decahydroquinolinemalonic acid diamide (24)

[0238]

[0239] Synthesis was performed similarly to Synthesis Example 16 from methyl malonate chloride (175 μl, 1.63 mmol) and trans-decahydroquinoline (500 mg, 3.59 mmol) to give methyl malonate N-decahydroquinoline amide, which was hydrolyzed to give malonic acid-N-decahydroquinoline amide.

[0240] MS(ESI+):m / z(%)226(100,[M+H] + ),248(85,[M+Na] + ).

[0241] This (190 mg, 0.843 mmol) was reacted with tert-butyl 3-aminopropylcarbamate (N-Boc-1,3-propylenediamine) (147 mg, 0.843 mmol) and TEA (236 μl, 1.686 mmol) in DMF (6 ml) with 685 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 50:50→40:60) gave the Boc-protected title compound (24a) (230 mg) in the form of a colorless oil.

[0242] MS(ESI+):m / z(%)382(100,[M+H] + ),404(80,[M+Na] + ),282(37,[M-Boc+H] + ),785(15,[2M+Na] + ).

[0243] The product thus obtained (0.603 mmol) was deprotected in a similar manner to give the hydrochloride (215 mg) of the title compound (24) in the form of a white powder.

[0244] MS(ESI+):m / z(%)282(100,[M+H) + ],304(30,[M+Na] + ),563(80,[2M+H] + ),585(60,[2M+Na] + ).

[0245] Synthesis Example 18

[0246] Preparation of N-(3-aminopropyl)-N'-decahydroisoquinolinemalonic acid diamide (25)

[0247]

[0248] Synthesis was performed similarly to Synthesis Example 17 from methyl malonate chloride (175 μl, 1.63 mmol) and decahydroisoquinoline (500 mg, 3.59 mmol) to give methyl malonate N-decahydroisoquinolineamide, which was hydrolyzed to give malonic acid-N-decahydroisoquinolineamide.

[0249] MS(ESI+):m / z(%)226(100,[M+H] + ),248(100,[M+Na] + ),473(36,[2M+Na] + ).

[0250] This (272 mg, 1.207 mmol) was reacted with tert-butyl 3-aminopropylcarbamate (N-Boc-1,3-propylenediamine) (210 mg, 1.207 mmol) and TEA (336 μl, 2.414 mmol) in DMF (10 ml) with 980 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (ethyl acetate) gave the Boc-protected title compound (25a) (306 mg) as a colorless oil.

[0251] MS(ESI+):m / z(%)382(100,[M+H] + ),404(100,[M+Na] + ),282(100,[M-Boc+H] + ),785(100,[2M+Na] + ).

[0252] The product thus obtained (0.786 mmol) was deprotected in a similar manner to give the hydrochloride (280 mg) of the title compound (25) in the form of a white powder.

[0253] MS(ESI+):m / z(%)282(100,[M+H]+),304(20,[M+Na] + ),563(40,[2M+H] + ),585(50,[2M+Na] + ).

[0254] Synthesis Example 19

[0255] Preparation of N-(3-aminopropyl)-N'-(pinan-10-yl)malonic acid diamide (26)

[0256]

[0257] Similarly to Synthesis Example 16, synthesis was carried out from methyl malonate chloride (250 μl, 2.33 mmol) and (-)-cis-(pinan-2-ylmethyl)amine (myrtanylamine, 10-pinanamine) (860 mg, 5.13 mmol) to obtain methyl malonate N-(pinan-10-yl)amide, which was hydrolyzed to obtain malonic acid-N-(pinan-10-yl)amide.

[0258] MS(ESI+):m / z(%)240(100,[M+H] + ),262(55,[M+Na] + ),501(10,[2M+Na] + ).MS(ESI-)m / z(%)238(100,[MH] - ),477(100,[2M-H] - ).

[0259] This (295 mg, 1.327 mmol) was reacted with tert-butyl 3-aminopropylcarbamate (N-Boc-1,3-propylenediamine) (231 mg, 1.327 mmol) and TEA (371 μl, 2.654 mmol) in DMF (7 ml) with 1.1 ml of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 50:50 → ethyl acetate) gave the Boc-protected title compound (26a) (278 mg) in the form of a colorless oil.

[0260] 1 H NMR (CDCl 3 ): δ=0.88(d,1H,J=8.5Hz),1.02(s,3H),1.18(s,3H),1.44(s,9H),1.45-1.50(m,1H),1.59-1.67(m,4H),1.84-1.98(m,5H),2.16- 2.24(m,1H),2.33-2.38(m,1H),3.16(s,2H),3.13-3.18(m,2H),3.24-3.34(m,4H),7.05(s,1H).MS(ESI+):m / z(%)396(100,[M+H] + ),296(100,[M-Boc+H] +),418(100,[M+Na]+),791(100,[2M+H] + ),813(100,[2M+Na] + ).

[0261] The product thus obtained (0.703 mmol) was deprotected in a similar manner to give the hydrochloride salt (260 mg) of the title compound (26) as a white solid.

[0262] 1 H NMR (DMSO-d 6 ): δ=0.84(d,1H,J=8.5Hz),0.99(s,3H),1.15(s,3H),1.38-1.45(m,1H) ,1.65-1.72(m,2H),1.79-1.91(m,5H),2.06-2.14(m,1H),2.29-2.34(m ,1H),2.75-2.83(m,2H),3.01(s,2H),3.03-3.08(m,2H),3.10-3.16(m, 2H),3.64-3.73(m,1H),7.82(brs,2H).MS(ESI+):m / z(%)296(100,[M+H] + ),318(90,[M+Na] + ),591(100,[2M+H] + ),613(100,[2M+Na] + ).

[0263] Synthesis Example 20

[0264] Preparation of N-(3-aminopropyl)-N'-tetrahydroquinolinemalonic acid diamide (108)

[0265]

[0266] Synthesis was performed similarly to Synthesis Example 17 from methyl malonate chloride (175 μl, 1.63 mmol) and 1,2,3,4-tetrahydroquinoline (478 mg, 3.59 mmol) to give methyl malonate N-tetrahydroquinoline amide, which was hydrolyzed to give malonic acid-N-tetrahydroquinoline amide.

[0267] MS(ESI+):m / z(%)220(100,[M+H] + ),242(100,[M+Na] + ),461(17,[2M+Na] + ).MS(ESI-)m / z(%)218(100,[MH]- ).

[0268] This (250 mg, 1.140 mmol) was reacted with tert-butyl 3-aminopropylcarbamate (N-Boc-1,3-propylenediamine) (199 mg, 1.140 mmol) and TEA (318 μl, 2.280 mmol) in DMF (10 ml) with 900 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 70:30→40:60) gave the Boc-protected title compound (108a) (230 mg) in the form of a colorless oil.

[0269] 1 H NMR (CDCl 3 ): δ=1.43(s,9H),1.67(quint,2H,J=6.7Hz),1.98(quint,2H,J=6.7Hz),2.72(brs,2H),3.16(q,2H,J=6.2Hz),3.34(q ,2H,J=6.4Hz),3.47(s,2H),3.82(t,2H,J=6.7Hz),5.00(brs,1H),7.09-7.23(m,4H).MS(ESI+):m / z(%)376(100,[M+H] + ),398(100,[M+Na] + ),276(100,[M-Boc+H] + ),773(100,[2M+Na] + ).MS(ESI-)m / z(%)374(100,[MH] - ).

[0270] The product thus obtained (275 mg, 0.732 mmol) was deprotected in a similar manner to give the hydrochloride (264 mg) of the title compound (108) in the form of a white powder.

[0271] 1 H NMR (DMSO-d 6 ): δ=1.67(quint,2H,J=7.1Hz), 1.87(quint,2H,J=6.6), 2.69(t,2H,J=6.6Hz), 2.78(q,2H,J=6.6Hz), 3.11(q,2H,6.5H z),3.67(s,2H),3.64-3.72(m,2H),7.09-7.20(m,4H),7.82(brs,2H),8.22(brs,1H).MS(ESI+):m / z(%)276(100,[M+H]+ ),298(8,[M+Na] + ),551(11,[2M+H] + ),573(18,[2M+Na] + ).

[0272] Synthesis Example 21

[0273] Preparation of malonyl adamantane-1-yl ester-N-(3-dimethylaminopropyl)amide (27)

[0274]

[0275] To a solution of adamantane-1-yl malonate (8) (350 mg, 1.469 mmol) and 3-aminopropyldimethylamine (150 mg, 1.469 mmol) in DMF (10 ml), triethylamine (TEA) (409 μl, 2.398 mmol) and 1.2 ml of a 1.6 M solution of propanephosphonic anhydride (T3P) in ethyl acetate were slowly added and stirred at room temperature for 1 h. The reaction mixture was then mixed with saturated brine for hydrolysis and extracted with ethyl acetate. The extract was purified by MgSO 4 Dry, filter and evaporate to dryness.The residue was purified by column chromatography (dichloromethane / ethyl acetate 50:50→ethyl acetate / methanol 70:30) to give the title compound (27) (400 mg) as a colorless oil.

[0276] 1 H NMR (CDCl 3 ): δ = 1.66 (m, 6H, adamantyl), 1.72 (quint, 2H, J = 6.77 Hz), 2.04-2.11 (m, 9H, adamantyl), 2.28 (s, 6H), 2.42 (t, 2H, J = 6.92 Hz), 3.19 (s, 2H), 3.45 (q, 2H, J = 6.30 Hz), 7.62 (brs, 1H). MS (ESI+): m / z (%) 323 (100, [M+H] + ),345(11,[M+Na] + ).

[0277] Synthesis Example 22

[0278] Preparation of malonyl adamantane-1-yl ester-N-(3-morpholinopropyl)amide (28)

[0279]

[0280] Analogously to Synthesis Example 21, from malonate adamantane-1-yl ester (8) (190 mg, 0.797 mmol), 3-morpholinylpropylamine (115 mg, 0.797 mmol) and triethylamine (TEA) (222 μl, 1.594 mmol) in DMF (5 ml), synthesized together with 650 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (ethyl acetate → ethyl acetate / methanol 90:10) gave the title compound (28) (250 mg) in the form of a colorless oil.

[0281] 1 H NMR (CDCl 3 ): δ = 1.65-1.74 (m, 8H), 2.10 (m, 6H, adamantyl), 2.17 (brs, 3H, adamantyl), 2.40-2.44 (m, 6H), 3.19 (s, 2H), 3.33-3.38 (m, 2H), 3.72 (t, 4H, J = 4.68 Hz), 7.61 (brs, 1H). MS (ESI+): m / z (%) 365 (100, [M+H] + ),387(20,[M+Na] + ),751(67,[2M+Na] + ).

[0282] Synthesis Example 23

[0283] Preparation of malonyl adamantane-1-yl ester-4-(3-trifluoromethylbenzyl)piperazinamide (29)

[0284]

[0285] Analogously to Synthesis Example 21, the product was synthesized from adamantane-1-yl malonate (8) (85 mg, 0.357 mmol), 1-(3-trifluoromethylbenzyl)piperazine (87 mg, 0.357 mmol) and triethylamine (TEA) (100 μl, 0.714 mmol) in DMF (2 ml) together with 290 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 90:10→50:50) gave the title compound (29) as a colorless oil (116 mg).

[0286] 1 H NMR (CDCl 3): δ = 1.65 (m, 6H, adamantyl), 2.10-2.17 (m, 9H, adamantyl), 2.45 (q, 4H, J = 4.61 Hz), 3.37 (s, 2H), 3.44 (t, 2H, J = 5.05 Hz), 3.57 (s, 2H), 3.66 (t, 2H, J = 5.05 Hz), 7.42-7.59 (m, 4H, H-aryl). MS (ESI+): m / z (%) 465 (100, [M+H] + ),487(42,[M+Na] + ),951(7,[2M+Na] + ).

[0287] synthesis Implementation Example 24

[0288] Preparation of malonyl adamantane-1-yl ester-4-(3-fluorobenzyl)piperazinamide (30)

[0289]

[0290] Analogously to Synthesis Example 21, the product was synthesized from adamantane-1-yl malonate (8) (127 mg, 0.533 mmol), 1-(3-fluorobenzyl)piperazine (104 mg, 0.533 mmol) and triethylamine (TEA) (149 μl, 1.07 mmol) in DMF (4 ml) together with 433 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 80:20→50:50) gave the title compound (30) (172 mg) in the form of a colorless oil.

[0291] 1 H NMR (CDCl 3 ): δ = 1.65 (m, 6H, adamantyl), 2.10-2.16 (m, 9H, adamantyl), 2.44 (q, 4H, J = 5.20 Hz), 3.37 (s, 2H), 3.43 (t, 2H, J = 5.00 Hz), 3.51 (s, 2H), 3.65 (t, 2H, J = 5.07 Hz), 6.93-6.98 (m, 1H, H-aryl), 7.05-7.08 (m, 2H, H-aryl), 7.25-7.30 (m, 1H, H-aryl). MS (ESI+): m / z (%) 415 (100, [M+H] + ),437(32,[M+Na] + ),851(10,[2M+Na] + ).

[0292] Synthesis Example 25

[0293] Preparation of malonyl adamantane-1-yl ester-4-(2-methoxyphenyl)piperazinamide (31)

[0294]

[0295] Analogously to Synthesis Example 21, the product was synthesized from adamantane-1-yl malonate (8) (100 mg, 0.420 mmol), 1-(2-methoxyphenyl)piperazine (81 mg, 0.420 mmol) and triethylamine (TEA) (341 μl, 0.546 mmol) in DMF (4 ml) together with 341 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 95:5→85:15) gave the title compound (31) (130 mg) in the form of white crystals.

[0296] 1 H NMR (CDCl 3 ): δ = 1.65 (m, 6H, adamantyl), 2.12-2.17 (m, 9H, adamantyl), 3.06 (dt, 4H, J = 14.37, 10.21 Hz), 3.42 (s, 2H), 3.62 (t, 2H, J = 5.04 Hz), 3.82 (t, 2H, J = 5.10 Hz), 3.88 (s, 3H), 6.88-6.95 (m, 3H, H-aryl), 7.02-7.06 (m, 1H, H-aryl). MS (ESI+): m / z (%) 413 (100, [M+H] + ),435(57,[M+Na] + ),847(38,[2M+Na] + ).

[0297] Synthesis Example 26

[0298] Preparation of malonic acid-N-(3,5-dimethyladamantan-1-yl)amide (32)

[0299]

[0300] Methyl malonate chloride (173 mg, 1.27 mmol) was dissolved in 10 ml of dichloromethane under an inert gas atmosphere and cooled to -20 °C, after which a solution of 3,5-dimethyl-1-adamantanamine (500 mg, 2.79 mmol) in 10 ml of dichloromethane was slowly added using a syringe. The reaction mixture was stirred at room temperature for 1.5 hours, after which the solvent was removed and the residue was redissolved in ethyl acetate. The organic phase was treated with 1N HCl and saturated NaHCO 3Wash with aqueous solution and MgSO 4 Dry, filter and evaporate to dryness. The residue is dissolved in THF and H 2 O (2:1), 6 ml of 1M NaOH aqueous solution was added thereto and stirred at room temperature. After 1 hour, THF was removed on a rotary evaporator, and the remaining aqueous solution was acidified with 2N HCl and extracted with ethyl acetate. The organic phase was purified by MgSO 4 Drying, filtration and evaporation to dryness gave the title compound (32) as white crystals (260 mg).

[0301] 1 H NMR (DMSO-d 6 ): δ=0.80(s,6H),1.08(s,2H),1.26(qd,4H,J=6.18,2.86Hz),1.55(s,4H),1.72(d,2H,J=2.97Hz),2.0 5(quint,1H,J=3.10Hz),3.04(s,2H),7.56(brs,1H),12.39(brs,1H).MS(ESI+):m / z(%)266(100,[M+H] + ),288(9,[M+Na] + ).

[0302] Synthesis Example 27

[0303] Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-(3-aminopropyl)malonic acid diamide (33)

[0304]

[0305] Similar to Synthesis Example 21, by malonic acid-N-(3,5-dimethyladamantan-1-yl)amide (32) (100 mg, 0.377 mmol), tert-butyl 3-aminopropylcarbamate (104) (66 mg, 0.377 mmol) and triethylamine (TEA) (105 μl, 0.754 mmol) in DMF (4 ml), synthesized together with 306 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 80:20→50:50) gave the Boc-protected title compound (33a) (110 mg) in the form of a colorless oil.

[0306] MS(ESI+):m / z(%)422(100,[M+H] + ),444(100,[M+Na]+ ),865(90,[2M+Na] + ).

[0307] The residue thus obtained (110 mg, 0.261 mmol) was dissolved in 30 ml of dichloromethane, after which 402 μl of trifluoroacetic acid (TFA) were added, and the mixture was stirred at room temperature for 4 h. Subsequently, 60 ml of saturated NaHCO 3 The organic phase was purified by MgSO 4 Drying, filtration and evaporation to dryness gave the title compound (33) (76 mg) as a yellow oil.

[0308] 1 H NMR (DMSO-d6): δ=0.79(s,6H),1.08-1.10(m,3H),1.22-1.31(m,4H),1.54(s,4H),1.65(quint,2H,J=6.86Hz),1.72(d,2 H,J=2.48Hz),2.05(m,1H),2.77(t,2H,J=7.30Hz),2.96(s,2H),3.11(q,2H,J=6.36Hz).MS(ESI+):m / z(%)322(100,[M+H] + ),344(76,[M+Na] + ),643(70,[2M+H] + ).

[0309] Synthesis Example 28

[0310] Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-(3-morpholinopropyl)malonic acid diamide (34)

[0311]

[0312] The synthesis was carried out similarly to Synthesis Example 21 from malonic acid-N-(3,5-dimethyl)adamantan-1-ylamide (32) (48 mg, 0.181 mmol), 3-morpholinylpropylamine (26 mg, 0.181 mmol) and triethylamine (TEA) (51 μl, 0.362 mmol) in DMF (2 ml) together with 147 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (ethyl acetate → ethyl acetate / methanol 85:15) gave the title compound (34) (50 mg) as a colorless oil.

[0313] 1 H NMR (CDCl 3): δ=0.84(s,6H),1.11-1.19(m,2H),1.25-1.39(m,4H),1.60-1.72(m,6H),1.83(d,2H,J=2.95Hz),2.14(quint,1H,J=3.15Hz),2.40-2. 44(m,6H),3.02(s,2H),3.34(q,2H,J=6.14Hz),3.71(t,4H,J=4.67Hz),6.64(brs,1H),7.64(brs,1H).MS(ESI+):m / z(%)392(100,[M+H] + ),414(24,[M+Na] + ),805(22,[2M+Na] + ).

[0314] Synthesis Example 29

[0315] Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-piperazinemalonic acid diamide (35)

[0316]

[0317] Similar to Synthesis Example 4, from malonic acid-N-(3,5-dimethyl)adamantan-1-ylamide (32) (100 mg, 0.358 mmol), 1-piperazinecarbamic acid tert-butyl ester (67 mg, 0.358 mmol) and triethylamine (100 μl, 0.716 mmol) in DMF (5 ml) and 291 μl of a 1.6 M solution of T3P in ethyl acetate, the synthesis was performed. The residue obtained after extraction with ethyl acetate was purified by column chromatography (dichloromethane / ethyl acetate 80:20→50:50) to give the Boc-protected title compound (35a) (150 mg) in the form of a colorless oil.

[0318] 1 H NMR (CDCl 3 ): δ=0.83(s,6H),1.10-1.19(m,2H),1.25-1.30(m,2H),1.35-1.39(m,2H),1.46(s,9H,H-Boc),1.59-1.68(m,5H),1.83(d,2H,J=2.68H z),2.13(quint,1H,J=3.15Hz),3.23(s,2H),3.40-3.47(m,4H),3.51-3.54(m,2H),3.58-3.61(m,2H).MS(ESI+):m / z(%)434(100,[M+H] +),456(100,[M+Na] + ),867(100,[2M+H] + ),889(100,[2M+Na] + ).

[0319] The thus obtained product (150 mg, 0.346 mmol) was deprotected similarly to Synthesis Example 4 to give the hydrochloride (125 mg) of the title compound (35) in the form of a white solid.

[0320] 1 H NMR (DMSO-d6): δ = 0.79 (s, 6H), 1.08 (s, 2H), 1.21-1.31 (m, 4H), 1.54 (s, 4H), 1.72 (s, 2H, J = 2.84Hz), 2.05 (m, 1H), 2.99(brs,2H),3.09(brs,2H),3.63-3.67(m,4H),7.66(brs,1H),9.30(brs,1H).MS(ESI+):m / z(%)334(100,[M+H] + ),356(31,[M+Na] + ),689(32,[2M+Na] + ).

[0321] Synthesis Example 30

[0322] Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-4-(3-trifluoromethylbenzyl)piperazinemalonic acid diamide (36)

[0323]

[0324] Analogously to Synthesis Example 21, malonic acid-N-(3,5-dimethyladamantan-1-yl)amide (32) (95 mg, 0.358 mmol), 1-(3-trifluoromethylbenzyl)piperazine (87 mg, 0.357 mmol) and triethylamine (TEA) (100 μl, 0.714 mmol) in DMF (2 ml) were synthesized together with 290 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 80:20→40:60) gave the title compound (36) (110 mg) in the form of a colorless oil.

[0325] 1 H NMR (CDCl 3): δ=0.84(s,6H),1.10-1.20(m,2H),1.25-1.30(m,2H),1.36-1.39(m,2H),1.64(q,4H,J=11.77Hz),1.83(brs,2H),2.13(qui nt,1H,J=3.13Hz),2.43(brs,4H),3.21(s,2H),3.56-3.65(m,6H),7.42-7.59(m,4H,H-aryl).MS(ESI+):m / z(%)492(100,[M+H] + ),514(85,[M+Na] + ),983(8,[2M+H] + ).

[0326] Synthesis Example 31

[0327] Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-4-(3-fluorobenzyl)piperazinemalonic acid diamide (37)

[0328]

[0329] The synthesis was carried out similarly to Example 21 from malonic acid-N-(3,5-dimethyladamantan-1-yl)amide (32) (95 mg, 0.358 mmol), 1-(3-fluorobenzyl)piperazine (70 mg, 0.358 mmol) and triethylamine (TEA) (100 μl, 0.716 mmol) in DMF (2 ml) together with 290 μl of a 1.6 M solution of T3P in ethyl acetate. The title compound (37) (107 mg) was purified by column chromatography (dichloromethane / ethyl acetate 50:50→ethyl acetate) to give a colorless oil.

[0330] 1 H NMR (CDCl 3 ): δ=0.84(s,6H),1.10-1.20(m,2H),1.25-1.30(m,2H),1.36-1.40(m,2H),1.64(q,4H,J=11.47Hz),1.83(d,2H,J=2.29Hz),2.13( quint,1H,J=3.15Hz),2.43(brs,4H),3.21(s,2H),3.51-3.63(m,6H),6.94-7.30(m,4H,H-Aryl).MS(ESI+):m / z(%)442(100,[M+H] + ),464(75,[M+Na] + ),905(100,[2M+Na]+ ).

[0331] Synthesis Example 32

[0332] Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-4-(2-methoxyphenyl)piperazinemalonic acid diamide (38)

[0333]

[0334] Analogously to Synthesis Example 21, malonic acid-N-(3,5-dimethyladamantan-1-yl)amide (32) (95 mg, 0.358 mmol), 1-(2-methoxyphenyl)piperazine (69 mg, 0.358 mmol) and triethylamine (TEA) (100 ml, 0.716 mmol) in DMF (2 ml) were synthesized together with 290 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 90:10→50:50) gave the title compound (38) (120 mg) in the form of a white foam.

[0335] 1 H NMR (CDCl 3 ): δ=0.84(s,6H),1.10-1.19(m,2H),1.26-1.29(m,2H),1.36-1.40(m,2H),1.61-1.70(m,5H),1.85(d,2H,J=2.76Hz),2.13(quint,1H,J=3 .15Hz),3.05(brs,4H),3.27(s,2H),3.74(brs,2H),3.82(brs,2H),3.88(s,3H),6.88-7.05(m,4H,H-aryl).MS(ESI+):m / z(%)440(100,[M+H] + ),462(77,[M+Na] + ),901(84,[2M+Na] + ).

[0336] Synthesis Example 33

[0337] Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-4-(4-methoxyphenyl)piperazinemalonic acid diamide (39)

[0338]

[0339] Analogously to Synthesis Example 21, malonic acid-N-(3,5-dimethyladamantan-1-yl)amide (32) (95 mg, 0.358 mmol), 1-(4-methoxyphenyl)piperazine (69 mg, 0.358 mmol) and triethylamine (TEA) (100 ml, 0.716 mmol) in DMF (2 ml) were synthesized together with 290 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 90:10→50:50) gave the title compound (38) (113 mg) in the form of a white solid.

[0340] 1 H NMR (CDCl 3 ): δ=0.84(s,6H),1.10-1.19(m,2H),1.26-1.29(m,2H),1.35-1.40(m,2H),1.60-1.69(m,5H),1.84(d,2H,J=2.75Hz),2.13(quint,1 H,J=3.14Hz),3.05(brs,4H),3.27(s,2H),3.77(s,3H),3.71-3.78(m,4H),6.84-6.91(m,4H,H-aryl).MS(ESI+):m / z(%)440(100,[M+H] + ),462(100,[M+Na] + ),901(100,[2M+Na] + ).

[0341] Synthesis Example 34

[0342] Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-4-(2,4-dimethoxyphenyl)piperazinemalonic acid diamide (40)

[0343]

[0344] Analogously to Synthesis Example 21, malonic acid-N-(3,5-dimethyladamantan-1-yl)amide (32) (85 mg, 0.320 mmol), 1-(2,4-dimethoxyphenyl)piperazine (71 mg, 0.320 mmol) and triethylamine (TEA) (90 ml, 0.640 mmol) in DMF (2 ml) were synthesized together with 240 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 90:10→40:60) gave the title compound (40) (120 mg) in the form of an off-white solid.

[0345] 1 H NMR (CDCl 3 ): δ=0.84(s,6H),1.10-1.20(m,2H),1.26-1.29(m,2H),1.35-1.40(m,2H),1.61-1. 70(m,5H),1.85(d,2H,J=2.71Hz),2.13(quint,1H,J=3.13Hz),2.96(brs,4H),3.26 (s,2H),3.71(brs,2H),3.78(s,3H),3.80(brs,2H),6.42(dd,2H,J=8.61Hz,2.55Hz ),6.49(d,2H,J=2.65Hz),6.82(d,2H,J=8.43Hz).MS(ESI+):m / z(%)470(100,[M+H] + ,492(87,[M+Na] + ),961(14,[2M+Na] + ).

[0346] Synthesis Example 35

[0347] Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-4-(2-morpholinyl-2-oxoethyl)piperazine malonic acid diamide (41)

[0348]

[0349] Analogously to Synthesis Example 21, malonic acid-N-(3,5-dimethyladamantan-1-yl)amide (32) (68 mg, 0.256 mmol), 1-morpholinyl-2-(piperazine-1-yl)ethanone (55 mg, 0.256 mmol) and triethylamine (TEA) (72 ml, 0.512 mmol) in DMF (2 ml) were synthesized together with 208 μl of a 1.6 M solution of T3P in ethyl acetate. Purification by column chromatography (dichloromethane / ethyl acetate 90:10) gave the title compound (41) (94 mg) in the form of a white foam.

[0350] 1 H NMR (CDCl 3): δ=0.84(s,6H),1.10-1.20(m,2H),1.25-1.29(m,2H),1.35-1.39(m,2H),1.64(q,4H,J=11.88Hz),1.83(d,2H,J=2.47Hz),2 .12(quint,1H,J=3.13Hz),2.63(brs,4H),3.22(s,2H),3.29(brs,2H),3.54-3.71(m,11H).MS(ESI+):m / z(%)461(100,[M+H] + ),483(100,[M+Na] + ),921(78,[2M+H] + ),943(58,[2M+Na] + ).

[0351] Example 1 to Example 41, Comparative Example 1 to Comparative Example 8 - Alzheimer's Disease Test

[0352] The potential suitability of the substances (1) to (41) and (101) to (108) prepared in the above-mentioned synthesis examples or purchased as agents for the treatment of Alzheimer's disease was tested in the screening assay mentioned above in a 96-well format using the transgenic C. elegans strain CL2659 in a standardized beta-amyloid test system using the nematode C. elegans (see Pretsch et al. above). For this purpose, worm larvae of the L3 stage were used at a density of 10-20 worms per well. The expression of beta-amyloid in the muscle cells was induced by increasing the temperature and the associated paralysis was recorded regularly until the end of the test after 48 hours. In this model, a significant delay of the larval paralysis caused by the neurotoxic active beta-amyloid should be observed for active substances at concentrations that reduce the expression of beta-amyloid compared to the control substances.

[0353] For this purpose, the above substances were tested in concentrations of 1 mg / ml, 100 μg / ml and 10 μg / ml compared to quercetin as a negative control. The test results are given in the following Table 1, wherein for the examples according to the invention the concentration ranges in μg / ml in which a delay in paralysis ("Alzheimer's disease") was observed are indicated, whereas for the comparative examples no effect was observed even at a concentration of 1 mg / ml.

[0354] Example 42 to Example 82, Comparative Example 9 to Comparative Example 16 - Parkinson's Disease Test

[0355] The same substances were tested for their potential suitability as agents for the treatment of Parkinson's disease in a 96-well format in a standardized α-synuclein test system using the nematode Caenorhabditis elegans (see Pretsch et al., supra) using the transgenic C. elegans strain NL5901. This worm strain expresses human α-synuclein in combination with green fluorescent protein (GFP) in all muscle cells. The GFP labeling allows the direct detection of the expressed protein levels for 24 to 48 hours using a multiplate reader. In this test model, a significant reduction in the expression of neurotoxic α-synuclein should be observed for active substances at concentrations that reduce α-synuclein expression compared to control substances.

[0356] For this purpose, the substances were tested again at concentrations of 1 mg / ml, 100 μg / ml and 10 μg / ml, but this time in comparison with the initially mentioned Parkinson's disease therapeutic agent Levodopa as a control. The test results are also given in the table on the next page, where again, for the examples according to the invention, the concentration ranges (in μg / ml) in which a reduction in the expression of alpha-synuclein can be detected are indicated ("Parkinson's disease"), while again, for the comparative examples in this test, no effect was observed even at a concentration of 1 mg / ml.

[0357]

[0358]

[0359]

[0360] The above test results clearly indicate that malonic acid and more than one of its derivatives are promising potential agents against both Alzheimer's and Parkinson's diseases, and possibly also against Huntington's disease or prion diseases.

[0361] The results obtained for various substitution patterns indicate that the active molecules can contain free carboxyl groups as well as salts, esters and substituted or unsubstituted amides without compromising their efficacy. Steric hindrance of the substituents also does not appear to play an important role, as both small groups (methyl, ethyl) and bulky groups (e.g., dimethyladamantyl or phenylpiperazinyl) show comparable efficacy.

[0362] As shown in the comparative examples, this efficacy is apparently based specifically on the presence of the unsubstituted malonic acid moiety, since neither dimethylmalonic acid (101) nor dimethylmalonate (102) proved to be effective. Substituents contained in the malonic acid derivatives effective according to the invention, such as (dimethyl)adamantyl or aminopropyl groups, have little effect on the degree of effectiveness, as demonstrated by the ineffectiveness of compounds (103) to (106). Of course, the ineffectiveness of tetrahydroquinolineamide (108) cannot be attributed to the presence of aromatic substituents, especially since all phenylpiperazineamide derivatives were also tested as positive, although the nonaromatic decahydroquinolineamide (24) and its isomer decahydroisoquinolineamide (25) were already effective at concentrations >10 μg / ml.

[0363] Furthermore, the 1:10 dilution factor chosen in the test series leaves a lot of room for the actual effective concentrations. For example, this does not exclude the possibility that a compound that is effective at concentrations >10 μg / ml and is therefore rated as "++" may only be effective at concentrations slightly below the 100 μg / ml limit, for example from 90 μg / ml upwards; while a compound that is only effective at concentrations above 100 μg / ml and is therefore rated as "+" may have an efficacy limit slightly above this limit. Such compounds may actually differ by only a few percentage points in their efficacy.

[0364] It should also be taken into account that the amount of the corresponding compound actually available in the test, i.e. the molar amount, depends of course on the molecular weight, which varies widely. For example, the molecular weights of free malonic acid (1) (104.06 g / mol) and malonic acid diamide (2) (102.09 g / mol) are only slightly more than one fifth of N-(3,5-dimethyladamantan-1-yl)-N'-4-(3-trifluoromethylbenzyl)piperazine malonic acid diamide (36) (491.60 g / mol). This factor of approximately 5 already anticipates almost half of the 1:10 dilution factor - and yet malonic acid itself and its three derivatives of relatively low molecular weight are only effective at concentrations >100 μg / ml, while compound (41) (i.e. 460.62 g / mol), which has a molecular weight second only to that of the aromatic compound (36), is already effective at >10 μg / ml.

[0365] Therefore, further studies to optimize the efficacy by means of further variations of the substitution pattern and dilution factor in the assay are the subject of current research by the inventors.

[0366] Thus, the present invention provides a group of compounds which, due to their efficacy in the nematode assay of C. elegans, should be useful for the treatment of neurodegenerative diseases such as Alzheimer's and Parkinson's diseases, as they are able to induce endogenous biosynthesis of metallothioneins even at very low concentrations.

Claims

1. Malonic acid and its derivatives of the following formula (I) for use as agents for promoting metallothionein biosynthesis in a patient: in, X and Y are each independently selected from -O - M + , -OR1 and -NR2R3, where M + R1, R2 and R3 are each independently selected from hydrogen and a saturated or unsaturated, linear, branched or cyclic hydrocarbon group having 1 to 25 carbon atoms, wherein one or more carbon atoms are optionally replaced by O or N, wherein R2 and R3 are optionally connected to each other and together with the nitrogen atom form a heterocyclic hydrocarbon group.

2. The compound for use according to claim 1, characterized in that In formula (I), both X and Y represent -O - M + , where each M + are independently selected from monovalent and multivalent metal cations, wherein preferably, M + represents an alkali metal ion or an alkaline earth metal ion, more preferably, M + Represents an alkali metal ion.

3. The compound for use according to claim 1 or 2, characterized in that R1, R2 and R3 are each independently selected from hydrogen and a saturated or unsaturated, linear, branched or cyclic hydrocarbon group having 1 to 15 carbon atoms.

4. The compound for use according to claim 3, characterized in that R1, R2 and R3 are each independently selected from hydrogen and a saturated or unsaturated hydrocarbon group having 1 to 10 carbon atoms.

5. The compound for use according to any one of claims 1 to 4, characterized in that The compound is selected from the following: malonic acid, malonic acid diamide, disodium malonate, dimethyl malonate, methyl malonate potassium salt, methyl malonate amide, N-(3-aminopropyl)malonic acid amide, adamantane-1-yl malonate, adamantane-1-yl ethyl malonate, adamantane-1-yl malonate-N-(3-aminopropyl)amide, adamantane-1-yl malonate-N-(3-aminopropyl)-N-methylamide, adamantane-1-yl malonate piperazinamide, adamantane-1-yl malonate-4-(4-aminobutyryl)piperazinamide, adamantane-1-yl malonate-4-aminopiperidinamide, adamantane-1-yl malonate-(4-(4-aminobutyryl)amino)piperidinamide, methyl malonate N-(adamantan-1-yl)amide, N-(Adamantane-1-yl)malonamide, N-adamantan-1-yl-N'-(3-aminopropyl)malonamide, N-adamantan-1-yl-N'-(3-aminopropyl)-N'-methylmalonamide, N-adamantan-1-yl-N'-piperazinemalonamide, N-adamantan-1-yl-N'-(4-(4-aminobutyryl)piperazine)malonamide, N-(3-aminopropyl)-N'-morpholinemalonamide, N-(3-aminopropyl)-N'-piperidinemalonamide, N-(3-aminopropyl)-N'-decahydroquinolinemalonamide, N-(3-aminopropyl)-N'-decahydroisoquinolinemalonamide, N-(3-aminopropyl)-N'-(6,6-dimethylbicyclo[3.1.1] heptane-2-ylmethyl) malonate diamide, malonate adamantane-1-yl ester-N-(3-dimethylaminopropyl)amide, malonate adamantane-1-yl ester-N-(3-morpholinylpropyl)amide, malonate adamantane-1-yl ester-4-(3-trifluoromethylbenzyl)piperazinamide, malonate adamantane-1-yl ester-4-(3-fluorobenzyl)piperazinamide, malonate adamantane-1-yl ester-4-(2-methoxyphenyl)piperazinamide, malonate-N-(3,5-dimethyl)adamantan-1-yl amide, N-(3,5-dimethyladamantan-1-yl)-N'-(3-aminopropyl) malonate diamide, N-(3,5-dimethyladamantan-1-yl)-N'-(3-morpholinylpropyl) malonate diamide, N-(3,5-dimethyladamantan-1-yl)-N'-(3-morpholinylpropyl) malonate diamide, )-N'-piperazine malonate diamide, N-(3,5-dimethyladamantan-1-yl)-N'-4-(3-trifluoromethylbenzyl)piperazine malonate diamide, N-(3,5-dimethyladamantan-1-yl)-N'-4-(3-fluorobenzyl)piperazine malonate diamide, N-(3,5-dimethyladamantan-1-yl)-N'-4-(2-methoxyphenyl)piperazine malonate diamide amide, N-(3,5-dimethyladamantan-1-yl)-N'-4-(4-methoxyphenyl)piperazine malonate diamide, N-(3,5-dimethyladamantan-1-yl)-N'-4-(2,4-dimethoxyphenyl)piperazine malonate diamide and N-(3,5-dimethyladamantan-1-yl)-N'-4-(2-morpholinyl-2-oxoethyl)piperazine malonate diamide.

6. The compound for use according to any one of claims 1 to 5, characterized in that Promoting metallothionein biosynthesis is useful for restoring metal homeostasis and thereby for treating neurodegenerative diseases.

7. The compound for use according to claim 6, characterized in that The neurodegenerative disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease and Huntington's disease.

8. The compound for use according to claim 7, characterized in that The neurodegenerative disease is Alzheimer's disease or Parkinson's disease.

9. A pharmaceutical composition for therapeutically treating the human or animal body by promoting metallothionein biosynthesis in the patient, the pharmaceutical composition comprising a compound as claimed in any one of claims 1 to 5, at least one pharmaceutically acceptable excipient and optionally one or more other pharmaceutically acceptable ingredients.

10. The pharmaceutical composition according to claim 9, characterized in that The pharmaceutical composition is used for treating neurodegenerative diseases.

11. The pharmaceutical composition according to claim 10, characterized in that The pharmaceutical composition is used for treating Alzheimer's disease or Parkinson's disease.

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