Metal constancy restoring agent containing malonic acid or its derivative

Abstract

The present invention relates to the use as a promoter for the biosynthesis of metallothionein in a patient of the following formula (I): JPEG2025522153000058.jpg1532 (I) (wherein X and Y are each independently selected from -O - M + , -OR1, and -NR2R3, M + is selected from monovalent or polyvalent metal anions, and R1, R2 and R3 are each independently selected from hydrogen and saturated or unsaturated straight-chain, branched or cyclic hydrocarbon radicals having 1 to 25 carbon atoms, one or more of the carbon atoms may be substituted by O or N, and R2 and R3 may combine to form a heterocyclic hydrocarbon radical together with the nitrogen atom), malonic acid and its derivatives; and a pharmaceutical composition containing a compound of formula (I).

Description

Technical Field

[0001] The present invention relates to a new drug that restores metal homeostasis, a drug with potential for neurodegenerative diseases.

Background Art

[0002] Latest Technology Neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and prion disease are age-related neuron diseases characterized by the accumulation of misfolded proteins and neuronal cell death. All of these diseases are currently one of the greatest challenges in medicine and occur worldwide.

[0003] Among approximately 50 million cases in 2020, Alzheimer's disease is a representative one of these diseases. It is confirmed that one person develops dementia every three seconds on Earth, and approximately 70% of them are Alzheimer's disease. Generally, Alzheimer's disease is a type of dementia that affects a patient's thinking, behavior, and memory. The progression of this disease is moderate, and neurons related to the learning part of the brain decrease. All current approaches to treating Alzheimer's disease only provide temporary symptom relief and do not inhibit or reverse the underlying disease mechanism. This is because they are mainly developed based on the amyloid cascade hypothesis and have not led to satisfactory treatment methods.

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

[0005] [Chemical formula]

[0006] WO2017 / 142855 A1 discloses derivatives of itaconic acid and malonic acid according to the following formulas I and II, respectively:

[0007] [Chemical formula]

[0008] (In the formula, R 1 to R 6 each independently represents hydrogen or an optionally substituted alkyl, alkenyl, or alkynyl group having any number of carbon atoms, and any substituent may, in addition to hydroxy, sometimes also include a vast range of (hetero)aromatic hydrocarbons. These definitions also include derivatives of succinic acid and maleic or fumaric acid, and itaconic acid (also known as methylene succinic acid), malonic acid, their dimethyl esters, as well as fumaric acid and 2-methylfumaric acid (mesaconic acid) are preferably mentioned.

[0009] These compounds are said to act as immunomodulators and, as a result, can be used in a variety of different types of diseases, such as neurodegenerative diseases like Alzheimer's disease, Parkinson's disease, Huntington's disease and prion diseases. In a preferred embodiment, the compounds are said to inhibit the expression of various proteins. However, in the example, only free malonic acid and itaconic acid were investigated for their effect as inhibitors of succinate dehydrogenase, which is said to be based on the structural similarity to the substrate succinic acid. The potential effectiveness in the treatment of neurodegenerative diseases cannot be inferred from this.

[0010] To overcome the lack of this treatment strategy and find other treatment options, research has been conducted on new targets for the treatment of these diseases. In 2008, Bush and Tanzi proposed the "metal hypothesis of Alzheimer’s disease" (A. Bush and R. E. Tanzi, “Therapeutics for Alzheimer’s disease based on the metal hypothesis”, Neurotherapeutics 5(3), 421-432 (2008)), suggesting that the breakdown of metal homeostasis is a major cause of neurodegenerative diseases, and thus drugs that restore metal homeostasis could provide a promising new treatment strategy. Subsequently, metal chelating agents have been repeatedly proposed as potential therapeutic agents for neurodegenerative diseases. For example, "Metal Chelation Therapy and Parkinson’s Disease: A Critical Review on the Thermo-dynamics of Complex Formation between Relevant Metal Ions and Promising or Estab-lished Drugs" by M. Tosato and A. Di Marco, Biomolecules 9(7), 269 (2019) lists approximately 800 substances that have been studied in the past few years as therapeutic agents for Parkinson's disease, among which malonic acid is also listed as a metal chelating agent; similarly, Acevedo et al. in "Redox active metals in neurodegenerative diseases", Biol. Inorg. Chem. 24(8), 1141-1157 (2019) clarify that metal chelating agents are a means to prevent the misfolding and aggregation of β-amyloid ("Aβ") and α-synuclein, which are one of the main causes of Alzheimer's disease and Parkinson's disease, etc.In a review by P.A. Adlard and A.I. Bush (“Metals and Alzheimer’s Disease: How Far Have We Come in the Clinic?”, J. Alzheimer’s Dis. 62(3), 1369-1379 (2018)), the chelating agent clioquinol (5-chloro-7-iodoquinolin-8-ol):

[0011]

Chem.

[0012] the same has been claimed.

[0013] In recent years, our working group has developed a new screening assay using transgenic strains of the nematode Caenorhabditis elegans, which is in the Rhabditida group. This is based on this new strategy of restoring metal homeostasis, specifically by upregulating metallothionein biosynthesis in the body: see Pretsch et al., “Prolongation of metallothionein induction combats As and α-synu-clein toxicity in aged transgenic Caenorhabditis elegans”, Sci. Rep. 10, 11707 (July 16, 2020). They explain that by prolonging the endogenous biosynthesis of metallothionein, a cytoplasmic protein that binds heavy metals, the expression of toxic β-amyloid (“Aβ”) and α-synuclein is suppressed, and the lifespan of the corresponding nematode larvae can be significantly extended. Emodin (1,3,8-trihydroxy-6-methylanthracene-9,10-dione) was identified as one of the agents to achieve this:

[0014]

Chem.

[0015] In a subsequent paper (D. Pretsch, “Abnormal metal homeostasis as a common drug target to combat neurodegenerative diseases”, Neural Regen. Res. 16(12), 2388-2389 (2021)), not only the effects of emodin but also those of clioquinol, as well as other effective (especially copper) chelates such as D-penicillamine (2-amino-3-mercapto-3-methylbutanoic acid) and diethyldithiocarbamic acid

[0016]

Chem.

[0017] are further stated not to be based solely on their chelating properties, as other effective (especially copper) chelates such as these failed in mouse test models related to Parkinson's disease. In this study, D-penicillamine showed no neuroprotective effect, but the neurotoxicity increased in the presence of diethyldithiocarbamic acid. In the C. elegans assay, the mechanisms of action of emodin and clioquinol should be unrelated to the binding of heavy metal ions by chelation, as they lead to an extension of the biosynthesis of metallothionein in the body.

[0018] Hama et al. (“Malonic acid suppresses lipopolysaccharide-induced BV2 microglia cell activation by inhibiting the p38 MAPK / NF-κB pathway”, Anim. Cells Syst. (Seoul) 25(2), 110-118 (2021)) also disclose malonic acid as a potential therapeutic agent for Alzheimer's disease and Parkinson's disease, as it has the ability to inhibit the expression of various cytokines such as interleukin, and as a result, can inhibit various signaling pathways involved in the occurrence of inflammation (e.g., the MAPK pathway).

[0019] Under such circumstances, an object of the present invention is to identify a chemical compound that is suitable as an agent for promoting the biosynthesis of metallothionein in vivo and can be an active agent with new potential for neurodegenerative diseases, using a new assay method based on C. elegans as a model organism.

Summary of the Invention

[0020] The present invention achieves this object by providing malonic acid of the following formula (I) and its derivatives for use as an agent for promoting the biosynthesis of metallothionein in a patient:

[0021]

Chemical formula

[0022] In the formula, X and Y are each independently selected from -O - M + , -OR1, and -NR2R3, where M + is selected from monovalent or polyvalent metal anions, and R1, R2, and R3 are each independently selected from hydrogen and saturated, unsaturated, straight-chain, branched, or cyclic hydrocarbon radicals having 1 to 25 carbon atoms, and one or more carbon atoms may be substituted with O or N, and R2 and R3 may optionally be bonded together to form a heterocyclic hydrocarbon radical together with the nitrogen atom.

[0023] Using the above-described assay method recently developed, the inventors have surprisingly found that both malonic acid itself of formula (I) and various derivatives thereof, namely both its esters, amides and salts, are effective as agents for promoting metallothionein biosynthesis in the C. elegans model organism and thereby restoring metal homeostasis, and as a result, are very likely to be suitable as agents for combating neurodegenerative diseases. Unsubstituted malonic acid has been disclosed several times as a therapeutic agent for neurodegenerative diseases due to its chelating properties, but in the nematode assay, it was not predictable whether either it or its derivatives would show a positive effect on the promotion of metallothionein biosynthesis in cells. This result is particularly surprising for long-chain derivatives, some of which are even substituted with aromatic compounds and some have relatively bulky structures, and the accessibility to the lone pair electrons of the central nucleophilic moiety C(=O)-CH2-C(=O) is limited, so the compatibility for the formation of metal chelates is significantly reduced.

[0024] However, this effectiveness was confirmed only for malonic acid derivatives in which the malonic acid moiety is not substituted at the central α-carbon atom, which is supported by the subsequent examples and comparative examples, and this is also surprising, clearly contradicting the idea that the reason for the effectiveness in this assay is only the ability of the compound to form a chelate with metal ions. This is also proven by the ineffectiveness of compound (108), N-(3-aminopropyl)-N'-tetrahydroquinoline malonic diamide, in Comparative Example 8, while all other aromatic substituted compounds show positive results.

[0025] In one group of embodiments (all of which are salts of malonic acid), at least one of X and Y of formula (I) is -O - M + but preferably both X and Y are -O - M + where M + is independently selected from monovalent and polyvalent metal anions, M +is preferably an alkali metal or alkaline earth metal ion, more preferably an alkali metal ion. Malonates containing only one carboxylate ion, as well as malonates in which both carboxyl groups are present as ionized carboxylates, have each been proven to be effective. However, in the latter case, since the manufacturing process is somewhat simpler, these are currently preferred in the present invention.

[0026] In the case of only a single malonate (in this case, only one of X and Y is -O - M + represents, and the other represents an ester group -OR1 or an amide group -NR2R3), and in both the case of diamides, diesters, and ester amides (in this case, both X and Y represent -OR1 or NR2R3), R1, R2, and R3 are preferably each independently selected from hydrogen and saturated or unsaturated linear, branched, or cyclic hydrocarbon radicals having 1 to 15 carbon atoms, and even more preferably selected from hydrogen and hydrocarbon radicals having 1 to 10 carbon atoms, because when the molecular weight of the compound is low, the amount of the active substance administered decreases.

[0027] In particular, the compounds according to the present invention are selected from the following: malonic acid (1), malonic acid diamide (2), disodium malonate (3), dimethyl malonate (4), potassium methyl malonate (5), methyl malonate amide (6), N-(3-aminopropyl) malonamide (7), adamantan-1-yl malonate (8), adamantan-1-yl ethyl malonate (9), adamantan-1-yl malonate-N-(3-aminopropyl)amide (10), adamantan-1-yl malonate-N-(3-aminopropyl)-N-methylamide (11), adamantan-1-yl malonate piperazine amide (12), adamantan-1-yl malonate-4-(4-aminobutyryl)piperazine amide (13), adamantan-1-yl malonate-4-aminopiperidine amide (14), adamantan-1-yl malonate-(4-(4-aminobutyryl)amino)piperidine amide (15), methyl malonate-N-(adamantan-1-yl)amide (16), N-(adamantan-1-yl)malonamide (17), N-adamantan-1-yl-N'-(3-aminopropyl)malonic diamide (18), N-adamantan-1-yl-N'-(3-aminopropyl)-N'-methylmalonic diamide (19), N-adamantan-1-yl-N'-piperazine malonic diamide (20), N-adamantan-1-yl-N'-(4-(4-aminobutyryl)piperazine)malonic diamide (21), N-(3-aminopropyl)-N'-morpholine malonic diamide (22), N-(3-aminopropyl)-N'-piperidine malonic diamide (23), N-(3-aminopropyl)-N'-decahydroquinoline malonic diamide (24), N-(3-aminopropyl)-N'-decahydroisoquinoline malonic diamide (25), N-(3-aminopropyl)-N'-(6,6-dimethylbicyclo[3.1.1) Diheptan-2-ylmalonamide (26), adamantan-1-yl malonate-N-(3-dimethylaminopropyl)amide (27), adamantan-1-yl malonate-N-(3-morpholinopropyl)amide (28), adamantan-1-yl malonate-4-(3-trifluoromethylbenzyl)piperazineamide (29), adamantan-1-yl malonate-4-(3-fluorobenzyl)piperazineamide (30), adamantan-1-yl malonate-4-(2-methoxyphenyl)piperazineamide (31), N-(3,5-dimethyl)adamantan-1-ylmalonamide (32), N-(3,5-dimethyladamantan-1-yl)-N'-(3-aminopropyl)malonamide (33), N-(3,5-dimethyladamantan-1-yl)-N'-(3-morpholinopropyl)malonamide (34), N-(3,5-dimethyladamantan-1-yl)-N'-piperazinylmalonamide (35), N-(3,5-dimethyladamantan-1-yl)-N'-4-(3-trifluoromethylbenzyl)piperazinylmalonamide (36), N-(3,5-dimethyladamantan-1-yl)-N'-4-(3-fluorobenzyl)piperazinylmalonamide (37), N-(3,5-dimethyladamantan-1-yl)-N'-4-(2-methoxyphenyl)piperazinylmalonamide (38), N-(3,5-dimethyladamantan-1-yl)-N'-4-(4-methoxyphenyl)piperazinylmalonamide (39), N-(3,5-dimethyladamantan-1-yl)-N'-4-(2,4-dimethoxyphenyl)piperazinylmalonamide (40), and N-(3,5-dimethyladamantan-1-yl)-N'-4-(2-morpholino-2-oxoethyl)piperazinylmalonamide (41).

[0028] In a preferred embodiment, promoting the biosynthesis of metallothionein in a patient restores metal homeostasis, thereby assisting in the treatment of the patient's neurodegenerative disease, which is particularly preferably selected from Alzheimer's disease, Parkinson's disease, and Huntington's disease, and in particular Alzheimer's disease can potentially be most effectively treated by restoring metal homeostasis according to the above "metal hypothesis", which according to Pretsch et al. (cited above) should be achievable by promoting metallothionein biosynthesis.

[0029] Accordingly, in a second aspect, the present invention also provides a pharmaceutical composition for therapeutically treating the body of a human or animal by promoting the biosynthesis of metallothionein in the patient, 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 components, the composition being useful preferably for the treatment of neurodegenerative diseases, more preferably Alzheimer's disease or Parkinson's disease, and particularly Alzheimer's disease.

Examples

[0030] The present invention will be described in more detail below by way of examples and comparative examples, which are for illustrative purposes only and should not be construed as limiting.

[0031] The compounds of the examples and comparative examples described below were either purchased commercially or prepared by the methods shown in the synthesis examples, and then tested for suitability as active agents against neurodegenerative diseases using the assay methods disclosed in the aforementioned "Pretsch et al.".

[0032] The test compounds were synthesized in a standard reaction sequence, and all starting products were either commercially available or pre-synthesized from commercially available reagents and used in the synthesis without further purification. Recorded at 400 MHz using an Avance AV400 spectrometer (Bruker) 1The property evaluation was performed using the 1H NMR spectrum and the mass spectrum recorded using LCMS-8040 (Shimadzu).

[0033] Specifically, in this first series of tests, the effectiveness of the following compounds was examined. Further tests are currently the subject of the inventors' research work.

[0034] As examples of the present invention, the following have been tested so far:

[0035]

Chemical formula

[0036]

Chemical formula

[0037]

Chemical formula

[0038]

Chemical formula

[0039]

Chemical formula

[0040]

Chemical formula

[0041]

Chemical formula

[0042]

Chemical formula

[0043] And also, the following compounds were examined as comparative examples:

[0044] [Chemical formula]

[0045] [Chemical formula]

[0046] [Chemical formula]

[0047] Among these, compounds (1) to (7) and (101) to (107) were obtained from commercially available products, and the other compounds (8) to (41) and (108) were prepared by the methods described below.

[0048] Synthesis Example 1 Preparation of adamantan-1-yl malonate (8)

[0049] [Chemical formula]

[0050] A toluene solution of 1.0 g (6.57 mmol) of adamantan-1-ol and 0.95 g (6.57 mmol) of 2,2-dimethyl-1,3-dioxane-4,6-dione was refluxed for 12 h, and then the reaction mixture was cooled to room temperature. Subsequently, 60 ml of saturated aqueous NaHCO3 solution was slowly added to cause 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, the organic phase was dried over MgSO4, filtered, and evaporated to dryness to obtain 1.5 g of the title compound as colorless crystals. 11H NMR (CD3OD): 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, [M-H] - ), 283 (47, [M+HCOO - ).

[0051] Synthesis Example 2 Preparation of adamantan-1-yl ethyl malonate (9)

[0052]

Chem.

[0053] To a solution of 35 mg of adamantan-1-yl ethyl 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 as a colorless liquid (35 mg). 1 1H NMR (CDCl3): δ = 1.25 (t, 3H, J = 7.14 Hz), 1.62 (m, 6H), 2.08 - 2.13 (m, 9H), 3.23 (s, 2H), 4.16 (q, 2H, J = 7.14 Hz). MS (ESI+): m / z (%) 267 (13, [M+H] + ), 284 (6, [M+H2O] + ), 289 (100, [M+Na] + ).

[0054] Synthesis Example 3 Preparation of adamantan-1-yl ethyl malonate-N-(3-aminopropyl)amide (10)

[0055]

Chem.

[0056] A solution of 22 mg (0.126 mmol) of tert-butyl 3-aminopropylcarbamate (N-Boc-1,3-propanediamine) (104), 30 mg (0.126 mmol) of adamantan-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 with 54 mg (0.126 mmol) of (1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylaminomorpholinocarbene hexafluorophosphate (COMU). After stirring for 2 h and warming to room temperature, the reaction mixture was diluted with ethyl acetate, washed with 1N HCl and saturated aqueous NaHCO3, dried over MgSO4, filtered and evaporated to dryness. The residue (47 mg) containing Boc-protected adamantan-1-yl malonate-N-(3-aminopropyl)amide (10a) was used in the next step without further purification. MS (ESI+): m / z (%) 395 (100, [M+H] + ), 417 (100, [M+Na] + ), 811 (100, [2M+Na] + ).

[0057] The residue thus obtained was dissolved in 8 ml of dichloromethane, 1.26 ml of 4N HCl in dioxane was added, and the mixture was stirred at room temperature for 2 h. Then, the volatile components were removed in vacuo to obtain the hydrochloride salt of the title compound (10) in the form of a yellow oil (34 mg). 1 H NMR (CDCl3): 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] + ).

[0058] Synthesis Example 4 Preparation of Adamantan-1-yl malonate-N-(3-aminopropyl)-N-methylamide (11)

[0059] [Chemical formula]

[0060] 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 a 1.6 M solution of propane phosphonic anhydride (T3P) in ethyl acetate, and the mixture was stirred at room temperature for 1 h. Next, the reaction mixture was mixed with saturated brine for hydrolysis and extracted with dichloromethane. The extract was dried over MgSO4, filtered, and evaporated to dryness. The residue was purified by column chromatography (dichloromethane / ethyl acetate 70:30) to obtain Boc-protected adamantan-1-yl malonate-N-(3-aminopropyl)-N-methylamide (11a) in the form of a colorless oil (370 mg). MS (ESI+): m / z (%) 409 (100, [M+H] + , 431 (100, [M+Na] + ), 309 (100, [M-Boc] + ), 839 (100, [2M+Na] + ).

[0061] The product thus obtained (0.96 mmol) was dissolved in 20 ml of dichloromethane, 9.6 ml of 4 N HCl in dioxane was added, and the mixture was stirred at room temperature for 1.5 h. Then, the volatile components were removed in vacuo to obtain the hydrochloride salt of the title compound (11) in the form of a yellow oil (260 mg). 11H NMR (DMSO-d6): 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] + ).

[0062] Synthesis Example 5 Preparation of Adamantan-1-yl Malonate Piperazine Amide (12)

[0063]

Chemical Structure

[0064] Synthesis was carried out in the same manner as in Synthesis Example 4 by adding adamantyl malonate (8) (150 mg, 0.630 mmol), 1-piperazinecarbamic acid tert-butyl ester (117 mg, 0.630 mmol), and TEA (176 μl, 1.26 mmol) to DMF (7 ml) together with 512 μl of a 1.6 M ethyl acetate solution of T3P. The residue was purified by column chromatography (dichloromethane / ethyl acetate 90:10 → 50:50) to obtain the Boc-protected title compound (12a) in the form of a white solid (232 mg). MS (ESI+): m / z (%) 407 (100, [M+H] + ), 429 (100, [M+Na] + ), 836 (100, [2M+Na] + ).

[0065] The product thus obtained (0.571 mmol) was deprotected in the same manner as in Synthesis Example 4 to obtain the hydrochloride salt of the title compound (12) in the form of a white solid (169 mg). 11H NMR (DMSO-d6): δ = 1.6 (t, 6H, J = 3.0 Hz), 2.03 (d, 6H, J = 3.0 Hz), 2.11 (brs, 3H), 3.05 (dt, 4H, J = 22.6, 5.2 Hz), 3.32 (brs, 1H), 3.48 (s, 2H), 3.63 (dt, 4H, J = 22.6, 5.2 Hz) MS (ESI+): m / z (%) 307 (100, [M+H] + ), 329 (25, [M+Na] + ), 635 (15, [2M+Na] + ).

[0066] Synthesis Example 6 Preparation of Adamantan-1-yl Malonate-4-(4-aminobutyryl)piperazine Amide (13)

[0067]

Chemical Structure

[0068] Synthesis was carried out in the same manner as in Synthesis Example 4. Adamantan-1-yl Malonate Piperazine Amide (12) (180 mg, 0.525 mmol), 4-(tert-butoxycarbonylamino)butanoic acid (107 mg, 0.525 mmol), and triethylamine (TEA) (219 μl, 1.575 mmol) were added to DMF (7 ml) together with 427 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (dichloromethane / ethyl acetate 80:20) gave the Boc-protected title compound (13a) in the form of a white solid (190 mg). MS (ESI+): m / z (%) 492 (100, [M+H] + ), 514 (100, [M+Na] + ), 392 (72, {M+H-Boc] + ).

[0069] The product thus obtained (0.386 mmol) was deprotected in the same manner as in Synthesis Example 4 to obtain the hydrochloride salt of the title compound (12) in the form of a white powder (151 mg). 1 H NMR (DMSO-d6): 1.59 (brs, 6H), 1.76 (quint, 2H, J = 7.4 Hz), 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] + ).

[0070] Synthesis Example 7 Preparation of Adamantan-1-yl malonate-4-aminopiperidine amide (14)

[0071]

Chemical formula

[0072] The synthesis was carried out in the same manner as in Synthesis Example 4 by adding adamantan-1-yl malonate (8) (150 mg, 0.630 mmol), tert-butyl piperidin-4-yl-carbamate (126 mg, 0.630 mmol) and triethylamine (TEA) (176 μl, 1.26 mmol) to DMF (7 ml) together with 512 μl of a 1.6 M ethyl acetate solution of T3P. The residue was purified by column chromatography (dichloromethane / ethyl acetate 90:10 → 80:20) to obtain the Boc-protected title compound (14a) in the form of a white solid (236 mg). MS (ESI+): m / z (%) 421 (100, [M+H] +), 443 (100, [M+Na] + ), 864 (100, [2M+Na] + ).

[0073] The product thus obtained (0.561 mmol) was deprotected in the same manner as in Synthesis Example 4 to obtain the hydrochloride of the title compound (14) in the form of a white powder (175 mg). MS (ESI+): m / z (%) 321 (100, [M+H] + ), 343 (7, [M+Na] + ), 641 (24, [2M+H] + ), 663 (95, [2M+Na] + ).

[0074] Synthesis Example 8 Preparation of Adamantan-1-yl malonate (4-(4-aminobutyryl)amino)piperidine amide (15)

[0075]

Chemical Structure

[0076] Synthesis was carried out in the same manner as in Synthesis Example 4 by adding adamantan-1-yl malonate-4-aminopiperidine amide (14) (140 mg, 0.392 mmol), 4-(tert-butoxycarbonylamino)butanoic acid (80 mg, 0.392 mmol) and triethylamine (TEA) (164 μl, 1.176 mmol) to DMF (7 ml) together with 319 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (dichloromethane / ethyl acetate 80:20 → ethyl acetate) gave the Boc-protected title compound (15a) in the form of a yellow oil (83 mg). MS (ESI+): m / z (%) 506 (100, [M+H] + ), 528 (100, [M+Na] + ), 406 (78, [M-Boc+H] + ).

[0077] The product thus obtained (0.164 mmol) was deprotected in the same manner as in Synthesis Example 4 to obtain the hydrochloride of the title compound (14) in the form of a yellow powder (80 mg). 1 H NMR (DMSO-d6): δ = 1.64 (dt, 3H, J=13.1, 2.8 Hz), 1.66 (dt, 3H, J=13.1, 2.8 Hz), 1.74 (sept, 3H, J=2.8 Hz), 1.80-1.87 (m, 6H), 2.09-2.13 (m, 6H), 2.16 (t, 2H, J=7.4 Hz), 2.65 (t, 2H, J=7.7 Hz), 3.04-3.13 (m, 2H), 3.35 (ddd, 2H, J=14.7, 6.8, 2.8 Hz), 3.55 (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] + ).

[0078] Synthesis Example 9 Preparation of methyl malonate-N-(adamantan-1-yl)amide (16)

[0079]

Chemical formula

[0080] Methyl malonyl chloride (322 μl, 3.005 mmol) was dissolved in 30 ml of dichloromethane under an inert gas atmosphere and cooled to -20 °C. Then, a 10 ml solution of 1-adamantanamine (107) (1.0 g, 6.612 mmol) in dichloromethane was slowly added using a syringe. The reaction mixture was stirred at room temperature for 1 h, quenched with saturated aqueous NH4Cl solution, and extracted with dichloromethane. The organic phase was dried over MgSO4, filtered, and evaporated to dryness to obtain the title compound as white crystals (760 mg). 1 1H NMR (CDCl3): 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] + ).

[0081] Synthesis Example 10 Preparation of N-(adamantan-1-yl)malonamide (17)

[0082]

Chemical Structure

[0083] To a solution of 750 mg (2.98 mmol) of methyl malonate-N-(adamantan-1-yl)amide (16) in 30 ml of THF / H2O (2:1) was added 6 ml of 1 M aqueous NaOH solution, and the mixture was stirred at room temperature. After 1 h, THF was removed by rotary evaporator, and the remaining aqueous solution was acidified with 2 N HCl and extracted with ethyl acetate. The organic phase was dried over MgSO4, filtered, and evaporated to dryness to obtain the title compound (17) as an off-white powder (670 mg). MS (ESI-): m / z (%) 236 (100, [M-H] - ), 473 (100, [2M-H] - ).

[0084] Synthesis Example 11 Preparation of N-adamantan-1-yl-N'-(3-aminopropyl)malonic diamide (18)

[0085]

Chem.

[0086] To a solution of N-(adamantan-1-yl)malonic amide (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) cooled to 0 °C in dichloromethane (8 ml) was added diisopropylcarbodiimide (DIC) (40 μl, 0.253 mmol). After stirring for 4 h and warming to room temperature, the reaction mixture was evaporated to dryness. The residue containing Boc-protected N-adamantan-1-yl-N'-(3-aminopropyl)malonic diamide (18a) (59 mg, 0.202 mmol) was used in the next step without further purification. MS (ESI+): m / z (%) 394 (100, [M+H] + ), 415 (100, [M+Na] + ), 294 (70, [M-BOC+H] + ), 787 (40, [2M+H] + ).

[0087] The residue thus obtained was dissolved in 6 ml of dichloromethane, 2 ml of 4N HCl in dioxane was added, and the mixture was stirred at room temperature for 2 h. Thereafter, the volatile components were removed in vacuo, and the hydrochloride of the title compound (18) was obtained in the form of a yellow oil (55 mg). 11H NMR (DMSO-d6): 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] + ).

[0088] Synthesis Example 12 Preparation of N-adamantan-1-yl-N'-(3-aminopropyl)-N'-methylmalonamide (19)

[0089]

Chemical Structure

[0090] Synthesis was carried out in the same manner as in Synthesis Example 4 by adding 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) to DMF (10 ml) together with 175 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (dichloromethane / ethyl acetate 70:30) gave the Boc-protected title compound (19a) in the form of a yellow oil (83 mg). MS (ESI+): m / z (%) 408 (100, [M+H] + ), 430 (100, [M+Na] + ), 308 (100, [M-BOC+H] + ), 815 (50, [2M+H] + ).

[0091] The product thus obtained (46 mg, 0.113 mmol) was deprotected in the same manner as in Synthesis Example 4 to obtain the hydrochloride salt of the title compound (19) in the form of a yellow oil (33 mg). 1 H NMR (DMSO-d6): 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] + ).

[0092] Synthesis Example 13 Preparation of N-adamantan-1-yl-N'-piperazine malonamide (20)

[0093]

Chemical formula

[0094] The synthesis was carried out in the same manner as in Synthesis Example 4 by adding commercially available 4-(tert-butoxycarbonyl)piperidine amide of malonic acid (250 mg, 0.918 mmol), 1-adamantanamine (107) (139 mg, 0.918 mmol) and triethylamine (TEA) (265 μl, 1.84 mmol) to DMF (17 ml) together with 750 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (dichloromethane / ethyl acetate 90:10→50:50) gave the Boc-protected title compound (20a) in the form of a white solid (260 mg). 11H NMR (CDCl3): δ = 1.46 (s, 9H), 1.67 (t, 7H, J = 3.9 Hz), 1.98 (d, 6H, J = 3.1 Hz), 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] + ).

[0095] The product thus obtained (0.647 mmol) was deprotected in the same manner as in Synthesis Example 4 to obtain the hydrochloride of the title compound (20) in the form of a white powder (192 mg). 1 1H NMR (DMSO-d6): 1.61 (brs, 6H), 1.9 (d, 6H, J = 2.95 Hz), 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] + ).

[0096] Synthesis Example 14 Preparation of N-adamantan-1-yl-N'-(4-(4-aminobutyryl)piperazine)malondiamide (21)

[0097]

Chemical Structure

[0098] The synthesis was carried out in the same manner as in Synthesis Example 4 by adding hydrochloride of N-adamantan-1-yl-N'-piperazine malonic diamide (20) (140 mg, 0.410 mmol), 4-(tert-butoxycarbonylamino)butanoic acid (83 mg, 0.410 mmol) and triethylamine (TEA) (171 μl, 1.23 mmol) to DMF (7 ml) together with 333 μl of 1.6 M T3P ethyl acetate solution. Purification by column chromatography (ethyl acetate / methanol 95:5) gave the Boc-protected title compound (21a) in the form of a white solid (180 mg). MS (ESI+): m / z (%) 491 (100, [M+H] + ), 513 (100, [M+Na] + ), 391 (38, [M-Boc+H] + ).

[0099] The product thus obtained (0.367 mmol) was deprotected in the same manner as in Synthesis Example 4 to give the hydrochloride of the title compound (21) in the form of a white powder (150 mg). 1 H NMR (DMSO-d6): 1.59 (brs, 6H), 1.77 (quint, 2H, J=6.4 Hz), 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).

[0100] Synthesis Example 15 Preparation of N-(3-aminopropyl)-N'-morpholine malonic diamide (22)

[0101]

Chemical formula

[0102] The synthesis was carried out in the same manner as in Synthesis Example 4 by adding commercially available N-morpholinomaleamic acid amide (90 mg, 0.520 mmol), tert-butyl 3-aminopropylcarbamate (N-Boc-1,3-propanediamine) (104) (91 mg, 0.520 mmol), and triethylamine (TEA) (145 μl, 1.04 mmol) to DMF (5 ml) together with 423 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (ethyl acetate / methanol 95:5) gave the Boc-protected title compound (22a) in the form of a yellow oil (90 mg). MS (ESI+): m / z (%) 330 (100, [M+H] + ), 352 (100, [M+Na] + ), 230 (100, [M-Boc+H] + ), 681 (100, [2M+Na] + ).

[0103] The product thus obtained (0.273 mmol) was deprotected in the same manner as in Synthesis Example 4 to give the hydrochloride salt of the title compound (22) in the form of a yellowish oil (76 mg). MS (ESI+): m / z (%) 230 (100, [M+H] + ), 252 (100, [M+Na] + ), 459 (11, [2M+H] + ), 481 (100, [2M+Na] + ).

[0104] Synthesis Example 16 Preparation of N-(3-aminopropyl)-N'-piperidinemaleamic diamide (23)

[0105]

Chemical formula

[0106] The synthesis was carried out in the same manner as the combination of Synthesis Examples 9, 10 and 4. First, similar to Synthesis Example 9, methyl malonate-N-piperidine amide was prepared from methyl malonate chloride (250 μl, 2.33 mmol) and piperidine (506 μl, 5.126 mmol) in 10 ml of dichloromethane at -20 °C. Next, similar to Synthesis Example 10, it was hydrolyzed with aqueous NaOH in THF / H2O (2:1) to obtain malonic acid-N-piperidine amide. MS (ESI+): m / z (%) 172 (100, [M+H] + ), 194 (50, [M+H] + ).

[0107] This (280 mg, 1.636 mmol) was then reacted in the same manner as Synthesis Example 4 with a mixture of tert-butyl 3-aminopropylcarbamate (N-Boc-1,3-propanediamine) (285 mg, 1.636 mmol) and TEA (456 μl, 3.272 mmol) in DMF (6 ml) together with 1.33 ml of a 1.6 M ethyl acetate solution of T3P. 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). 1 H NMR (CDCl3): δ = 1.43 (s, 9H), 1.53 - 1.60 (m, 4H), 1.63 - 1.68 (m, 5H), 3.14 (q, 2H, J = 6.5 Hz), 3.31 (s, 2H), 3.33 (q, 2H, J = 6.5 Hz), 3.45 (t, 2H, J = 4.8 Hz), 3.56 (t, 2H, J = 4.8 Hz), 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] + ).

[0108] The product thus obtained (1.13 mmol) was deprotected in the same manner as in Synthesis Example 4 to obtain the hydrochloride of the title compound (23) in the form of a white powder (330 mg). 1 H NMR (DMSO-d6): δ = 1.37-1.43 (m, 2H), 1.45-1.50 (m, 2H), 1.53-1.59 (m, 2H), 1.69 (quint, 2H, J=8.3 Hz), 2.75-2.83 (m, 2H), 3.12 (q, 2H, 4.8 Hz), 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] + ).

[0109] Synthesis Example 17 Preparation of N-(3-aminopropyl)-N'-dehydroquinoline malonamide (24)

[0110]

Chemical formula

[0111] The synthesis was carried out in the same manner as in Synthesis Example 16. Methyl malonyl chloride (175 μl, 1.63 mmol) and trans-dehydroquinoline (500 mg, 3.59 mmol) were used to prepare methyl malonyl-N-dehydroquinoline amide, which was then hydrolyzed to obtain malonyl-N-dehydroquinoline amide. MS (ESI+): m / z (%) 226 (100, [M+H] + ), 248 (85, [M+Na] + ).

[0112] This (190 mg, 0.843 mmol) was reacted with a solution prepared by adding tert-butyl 3-aminopropylcarbamate (N-Boc-1,3-propanediamine) (147 mg, 0.843 mmol) and TEA (236 μl, 1.686 mmol) to DMF (6 ml) together with 685 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (dichloromethane / ethyl acetate 50:50 → 40:60) gave the Boc-protected title compound (24a) in the form of a colorless oil (230 mg). MS (ESI+): m / z (%) 382 (100, [M+H] + ), 404 (80, [M+Na] + ), 282 (37, [M-Boc+H] + ), 785 (15, [2M+Na] + ).

[0113] The product thus obtained (0.603 mmol) was deprotected in a similar manner to give the hydrochloride salt of the title compound (24) in the form of a white powder (215 mg). MS (ESI+): m / z (%) 282 (100, [M+H) + , 304 (30, [M+Na] + ), 563 (80, [2M+H] + ), 585 (60, [2M+Na] + ).

[0114] Synthesis Example 18 Preparation of N-(3-aminopropyl)-N'-dehydroisoquinoline diamide malonate (25)

[0115]

Chemical formula

[0116] The synthesis was carried out in the same manner as in Synthesis Example 17. Methyl malonate chloride (175 μl, 1.63 mmol) and decahydroisoquinoline (500 mg, 3.59 mmol) were used to form methyl malonate-N-decahydroisoquinoline amide, which was then hydrolyzed to obtain malonic acid-N-decahydroisoquinoline amide. MS (ESI+): m / z (%) 226 (100, [M+H] + ), 248 (100, [M+Na] + ), 473 (36, [2M+Na] + ).

[0117] This (272 mg, 1.207 mmol) was reacted with a mixture of tert-butyl 3-aminopropylcarbamate (N-Boc-1,3-propanediamine) (210 mg, 1.207 mmol) and TEA (336 μl, 2.414 mmol) in DMF (10 ml) together with 980 μl of 1.6 M T3P ethyl acetate solution. Purification by column chromatography (ethyl acetate) gave the Boc-protected title compound (25a) in the form of a colorless oil (306 mg). MS (ESI+): m / z (%) 382 (100, [M+H] + ), 404 (100, [M+Na] + ), 282 (100, [M-Boc+H] + ), 785 (100, [2M+Na] + ).

[0118] The product thus obtained (0.786 mmol) was deprotected in the same manner to give the hydrochloride salt of the title compound (25) in the form of a white powder (280 mg). MS (ESI+): m / z (%) 282 (100, [M+H]+), 304 (20, [M+Na] + ), 563 (40, [2M+H] + ), 585 (50, [2M+Na] + ).

[0119] Synthesis Example 19 Preparation of N-(3-aminopropyl)-N'-(pinan-10-yl)malonamide (26)

[0120] [Chemical formula]

[0121] The synthesis was carried out in the same manner as in Synthesis Example 16. From methyl malonyl chloride (250 μl, 2.33 mmol) and (-)-cis-myrtanilamine (10-pinanamine) (860 mg, 5.13 mmol), methyl malonyl-N-(pinan-10-yl)amide was prepared, and this was hydrolyzed to obtain malonic acid-N-(pinan-10-yl)amide. MS (ESI+): m / z (%) 240 (100, [M+H] + ), 262 (55, [M+Na] + ), 501 (10, [2M+Na] + ). MS (ESI-) m / z (%) 238 (100, [M-H] - ), 477 (100, [2M-H] - ).

[0122] This (295 mg, 1.327 mmol) was reacted with tert-butyl 3-aminopropylcarbamate (N-Boc-1,3-propanediamine) (231 mg, 1.327 mmol) and TEA (371 μl, 2.654 mmol) in DMF (7 ml) together with 1.1 ml of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (dichloromethane / ethyl acetate 50:50 → ethyl acetate) gave the Boc-protected title compound (26a) in the form of a colorless oil (278 mg). 11H NMR (CDCl3): δ = 0.88 (d, 1H, J = 8.5 Hz), 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] + ).

[0123] The product thus obtained (0.703 mmol) was deprotected in the same manner to give the hydrochloride salt of the title compound (26) in the form of a white solid (260 mg). 1 1H NMR (DMSO - d6): δ = 0.84 (d, 1H, J = 8.5 Hz), 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] + ).

[0124] Synthesis Example 20 Preparation of N-(3-aminopropyl)-N'-tetrahydroquinoline malonic diamide (108)

[0125]

Chemical formula

[0126] The synthesis was carried out in the same manner as in Synthesis Example 17. Methyl malonate chloride (175 μl, 1.63 mmol) and 1,2,3,4-tetrahydroquinoline (478 mg, 3.59 mmol) were used to prepare methyl malonate-N-tetrahydroquinoline amide, which was then hydrolyzed to obtain malonic acid-N-tetrahydroquinoline amide. MS (ESI+): m / z (%) 220 (100, [M+H] + ), 242 (100, [M+Na] + ), 461 (17, [2M+Na] + ). MS (ESI-) m / z (%) 218 (100, [M-H] - ).

[0127] This (250 mg, 1.140 mmol) was reacted with tert-butyl 3-aminopropylcarbamate (N-Boc-1,3-propanediamine) (199 mg, 1.140 mmol) and TEA (318 μl, 2.280 mmol) in DMF (10 ml) together with 900 μl of 1.6 M T3P ethyl acetate solution. Purification by column chromatography (dichloromethane / ethyl acetate 70:30 → 40:60) gave the Boc-protected title compound (108a) in the form of a colorless oil (230 mg). 11H NMR (CDCl3): δ = 1.43 (s, 9H), 1.67 (quint, 2H, J=6.7 Hz), 1.98 (quint, 2H, J=6.7 Hz), 2.72 (brs, 2H), 3.16 (q, 2H, J=6.2 Hz), 3.34 (q, 2H, J=6.4 Hz), 3.47 (s, 2H), 3.82 (t, 2H, J=6.7 Hz), 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, [M - H] - ).

[0128] The product thus obtained (275 mg, 0.732 mmol) was deprotected in the same manner to obtain the hydrochloride of the title compound (108) in the form of a white powder (264 mg). 1 1H NMR (DMSO - d6): δ = 1.67 (quint, 2H, J=7.1 Hz), 1.87 (quint, 2H, J=6.6), 2.69 (t, 2H, J=6.6 Hz), 2.78 (q, 2H, J=6.6 Hz), 3.11 (q, 2H, 6.5 Hz), 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] + ).

[0129] Synthesis Example 21 Preparation of Adamantan-1-yl Malonate-N-(3-dimethylaminopropyl)amide (27)

[0130]

Chem.

[0131] To a solution of adamantan-1-yl malonate (8) (350 mg, 1.469 mmol) and 3-aminopropyldimethylamine (150 mg, 1.469 mmol) in DMF (10 ml) were slowly added triethylamine (TEA) (409 μl, 2.398 mmol) and 1.2 ml of a 1.6 M solution of propane phosphonic anhydride (T3P) in ethyl acetate, and the mixture was stirred at room temperature for 1 h. Next, the reaction mixture was mixed with saturated brine for hydrolysis and extracted with ethyl acetate. The extract was dried over MgSO4, filtered, and evaporated to dryness. The residue was purified by column chromatography (dichloromethane / ethyl acetate 50:50 → ethyl acetate / methanol 70:30) to obtain the title compound (27) in the form of a colorless oil (400 mg). 1 H NMR (CDCl3): 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] + ).

[0132] Synthesis Example 22 Preparation of Adamantan-1-yl Malonate-N-(3-morpholinopropyl)amide (28)

[0133]

Chem.

[0134] The synthesis was carried out in the same manner as in Synthesis Example 21. Adamantane-1-yl malonate (8) (190 mg, 0.797 mmol), 3-morpholinopropylamine (115 mg, 0.797 mmol), and triethylamine (TEA) (222 μl, 1.594 mmol) were added to DMF (5 ml) together with 650 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (ethyl acetate → ethyl acetate / methanol 90:10) gave the title compound (28) in the form of a colorless oil (250 mg). 1 H NMR (CDCl3): 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] + ).

[0135] Synthesis Example 23 Preparation of Adamantane-1-yl Malonate-4-(3-(Trifluoromethyl)Benzyl)Piperazine Amide (29)

[0136]

Chemical Structure

[0137] The synthesis was carried out in the same manner as in Synthesis Example 21. 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) were added to DMF (2 ml) together with 290 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (dichloromethane / ethyl acetate 90:10 → 50:50) gave the title compound (29) in the form of a colorless oil (116 mg). 1 H NMR (CDCl3): 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] + ).

[0138] Synthesis Example 24 Preparation of Adamantane-1-yl Malonate-4-(3-fluorobenzyl)piperazine Amide (30)

[0139]

Chemical Structure

[0140] The synthesis was carried out in the same manner as in Synthesis Example 21. 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) were added to DMF (4 ml) together with 433 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (dichloromethane / ethyl acetate 80:20 → 50:50) gave the title compound (30) in the form of a colorless oil (172 mg). 1 H NMR (CDCl3): 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] + ).

[0141] Synthesis Example 25 Preparation of Adamantane-1-yl Malonate-4-(2-Methoxyphenyl)Piperazine Amide (31)

[0142]

Chemical Structure

[0143] The synthesis was carried out in the same manner as in Synthesis Example 21. 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) were added to DMF (4 ml) together with 341 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (dichloromethane / ethyl acetate 95:5 → 85:15) gave the title compound (31) in the form of white crystals (130 mg). 1 H NMR (CDCl3): 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] + ).

[0144] Synthesis Example 26 Preparation of N-(3,5-dimethyladamantan-1-yl)malonamide (32)

[0145]

Chemical Structure

[0146] Methyl malonyl chloride (173 mg, 1.27 mmol) was dissolved in 10 ml of dichloromethane under an inert gas atmosphere and cooled to -20 °C. Then, a 10 ml dichloromethane solution of 3,5-dimethyl-1-adamantanamine (500 mg, 2.79 mmol) was slowly added using a syringe. After stirring the reaction mixture at room temperature for 1.5 h, the solvent was removed, and the residue was redissolved in ethyl acetate. The organic phase was washed with 1N HCl and saturated aqueous NaHCO3, dried over MgSO4, filtered, and evaporated to dryness. The residue of methyl malonate-N-(3,5-dimethyladamantan-1-yl)amide was dissolved in a mixture of THF and H2O (2:1), and 6 ml of 1M aqueous NaOH was added thereto, followed by stirring at room temperature. After 1 h, THF was removed using a rotary evaporator, the remaining aqueous solution was acidified with 2N HCl, and extracted with ethyl acetate. The organic phase was dried over MgSO4, filtered, and evaporated to dryness to obtain the title compound (32) in the form of white crystals (260 mg). 1 1H NMR (DMSO-d6): δ = 0.80 (s, 6H), 1.08 (s, 2H), 1.26 (qd, 4H, J = 6.18, 2.86 Hz), 1.55 (s, 4H), 1.72 (d, 2H, J = 2.97 Hz), 2.05 (quint, 1H, J = 3.10 Hz), 3.04 (s, 2H), 7.56 (brs, 1H), 12.39 (brs, 1H). MS (ESI+): m / z (%) 266 (100, [M+H] + ), 288 (9, [M+Na] + ).

[0147] Synthesis Example 27 Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-(3-aminopropyl)malonamide (33)

[0148]

Chemical Structure

[0149] The synthesis was carried out in the same manner as in Synthesis Example 21. 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) were added to DMF (4 ml) together with 306 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (dichloromethane / ethyl acetate 80:20 → 50:50) gave the title compound (33a) in the form of a colorless oil (110 mg). MS (ESI+): m / z (%) 422 (100, [M+H] + ), 444 (100, [M+Na] + ), 865 (90, [2M+Na] + ).

[0150] The residue thus obtained (110 mg, 0.261 mmol) was dissolved in 30 ml of dichloromethane, then 402 μl of trifluoroacetic acid (TFA) was added, and the mixture was stirred at room temperature for 4 h. Subsequently, 60 ml of saturated aqueous NaHCO3 was added, and the mixture was extracted with dichloromethane. The organic phase was dried over MgSO4, filtered, and evaporated to dryness to give the title compound (33) in the form of a yellowish oil (76 mg). 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 = 6.86 Hz), 1.72 (d, 2H, J = 2.48 Hz), 2.05 (m, 1H), 2.77 (t, 2H, J = 7.30 Hz), 2.96 (s, 2H), 3.11 (q, 2H, J = 6.36 Hz). MS (ESI+): m / z (%) 322 (100, [M+H] + ), 344 (76, [M+Na] + ), 643 (70, [2M+H] + ).

[0151] Synthesis Example 28 Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-(3-morpholinopropyl)malonic diamide (34)

[0152]

Chemical Structure

[0153] The synthesis was carried out in the same manner as in Synthesis Example 21. Malonic acid-N-(3,5-dimethyl)adamantan-1-ylamide (32) (48 mg, 0.181 mmol), 3-morpholinopropylamine (26 mg, 0.181 mmol) and triethylamine (TEA) (51 μl, 0.362 mmol) were added to DMF (2 ml) together with 147 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (ethyl acetate → ethyl acetate / methanol 85:15) gave the title compound (34) in the form of a colorless oil (50 mg). 1 H NMR (CDCl3): 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 = 2.95 Hz), 2.14 (quint, 1H, J = 3.15 Hz), 2.40 - 2.44 (m, 6H), 3.02 (s, 2H), 3.34 (q, 2H, J = 6.14 Hz), 3.71 (t, 4H, J = 4.67 Hz), 6.64 (brs, 1H), 7.64 (brs, 1H). MS (ESI+): m / z (%) 392 (100, [M+H] + ), 414 (24, [M+Na] + ), 805 (22, [2M+Na] + ).

[0154] Synthesis Example 29 Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-piperazine malonic diamide (35)

[0155] [Chem.]

[0156] The synthesis was carried out in the same manner as in Synthesis Example 4. Malonic acid - N-(3,5-dimethyl)adamantan-1-ylamide (32) (100 mg, 0.358 mmol), tert-butyl 1-piperazinecarbamate (67 mg, 0.358 mmol) and triethylamine (100 μl, 0.716 mmol) were added to DMF (5 ml) together with 291 μl of a 1.6 M ethyl acetate solution of T3P. The residue obtained after extraction with ethyl acetate was purified by column chromatography (dichloromethane / ethyl acetate 80:20 → 50:50) to obtain the Boc-protected title compound (35a) in the form of a colorless oil (150 mg). 1 H NMR (CDCl3): 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 = 2.68 Hz), 2.13 (quint, 1H, J = 3.15 Hz), 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] + ).

[0157] The product thus obtained (150 mg, 0.346 mmol) was deprotected in the same manner as in Synthesis Example 4 to obtain the hydrochloride salt of the title compound (35) in the form of a white solid (125 mg). 11H 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 = 2.84 Hz), 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] + ).

[0158] Synthesis Example 30 Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-4-(3-trifluoromethylbenzyl)piperazine diamide malonate (36)

[0159]

Chemical Structure

[0160] The synthesis was carried out in the same manner as in 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) were added to DMF (2 ml) together with 290 μl of a 1.6 M T3P ethyl acetate solution. Purification by column chromatography (dichloromethane / ethyl acetate 80:20 → 40:60) gave the title compound (36) in the form of a colorless oil (110 mg). 11H NMR (CDCl3): 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=11.77 Hz), 1.83 (brs, 2H), 2.13 (quint, 1H, J=3.13 Hz), 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] + ).

[0161] Synthesis Example 31 Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-4-(3-fluorobenzyl)piperazine diamide malonate (37)

[0162]

Chemical Structure

[0163] The synthesis was carried out in the same manner as in Synthesis Example 21. 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) were added to DMF (2 ml) together with 290 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (dichloromethane / ethyl acetate 50:50 → ethyl acetate) gave the title compound (37) in the form of a colorless oil (107 mg). 11H NMR (CDCl3): 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 = 11.47 Hz), 1.83 (d, 2H, J = 2.29 Hz), 2.13 (quint, 1H, J = 3.15 Hz), 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] + ).

[0164] Synthesis Example 32 Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-4-(2-methoxyphenyl)piperazine diamide malonate (38)

[0165]

Chemical Structure

[0166] The synthesis was carried out in the same manner as in 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 μl, 0.716 mmol) were added to DMF (2 ml) together with 290 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (dichloromethane / ethyl acetate 90:10 → 50:50) gave the title compound (38) in the form of white foam (120 mg). 11H NMR (CDCl3): 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 = 2.76 Hz), 2.13 (quint, 1H, J = 3.15 Hz), 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] + ).

[0167] Synthesis Example 33 Preparation of N-(3,5 - dimethyladamantan - 1 - yl)-N’ - 4-(4 - methoxyphenyl)piperazine diamide malonic acid (39)

[0168]

Chemical Structure

[0169] The synthesis was carried out in the same manner as in 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) were added to DMF (2 ml) together with 290 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (dichloromethane / ethyl acetate 90:10 → 50:50) gave the title compound (38) in the form of a white solid (113 mg). 11H NMR (CDCl3): 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 = 2.75 Hz), 2.13 (quint, 1H, J = 3.14 Hz), 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] + ).

[0170] Synthesis Example 34 Preparation of N-(3,5-dimethyladamantan-1-yl)-N’-4-(2,4-dimethoxyphenyl)piperazine diamide malonate (40)

[0171]

Chemical Structure

[0172] The synthesis was carried out in the same manner as in 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) were added to DMF (2 ml) together with 240 μl of a 1.6 M ethyl acetate solution of T3P. Purification by column chromatography (dichloromethane / ethyl acetate 90:10 → 40:60) gave the title compound (40) in the form of a white solid (120 mg). 11H NMR (CDCl3): 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 = 2.71 Hz), 2.13 (quint, 1H, J = 3.13 Hz), 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 = 8.61 Hz, 2.55 Hz), 6.49 (d, 2H, J = 2.65 Hz), 6.82 (d, 2H, J = 8.43 Hz). MS (ESI+): m / z (%) 470 (100, [M+H] + , 492 (87, [M+Na] + ), 961 (14, [2M+Na] + ).

[0173] Synthesis Example 35 Preparation of N-(3,5-dimethyladamantan-1-yl)-N'-4-(2-morpholino-2-oxoethyl)piperazine diamide malonate (41)

[0174]

Chemical Structure

[0175] The synthesis was carried out in the same manner as in Synthesis Example 21. Malonic acid - N-(3,5-dimethyladamantan-1-yl)amide (32) (68 mg, 0.256 mmol), 1-morpholino-2-(piperazin-1-yl)ethanone (55 mg, 0.256 mmol) and triethylamine (TEA) (72 μl, 0.512 mmol) were added to DMF (2 ml) together with 208 μl of 1.6 M T3P ethyl acetate solution. Purification by column chromatography (dichloromethane / ethyl acetate 90:10) gave the title compound (41) in the form of white foam (94 mg). 11H NMR (CDCl3): 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 = 11.88 Hz), 1.83 (d, 2H, J = 2.47 Hz), 2.12 (quint, 1H, J = 3.13 Hz), 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] + ).

[0176] Examples 1 to 41, Comparative Examples 1 to 8 - Alzheimer's Test Substances (1) to (41) and (101) to (108) prepared in the above synthesis examples or purchased were tested for potential suitability as therapeutic agents for Alzheimer's disease in a 96-well procedure in a standardized β-amyloid test system using the transgenic C. elegans CL2659 strain in the first-described screening assay using Caenorhabditis elegans (Pretsch et al., supra). For this purpose, L3-stage nematode larvae were used at a density of 10 - 20 per well. Expression of β-amyloid in muscle cells was induced by a temperature increase, and the associated paralysis was regularly recorded until the end of the test after 48 hours. In this model, using an active substance at a concentration that reduces the expression of β-amyloid compared to a control substance, a significant delay in the paralysis of larvae caused by neurotoxic active β-amyloid should be observed.

[0177] For this purpose, the above-mentioned substance was tested at concentrations of 1 mg / ml, 100 μg / ml, and 10 μg / ml, compared with quercetin as a negative control. The test results are shown in Table 1 below, and the concentration ranges (μg / ml) in which a delay in paralysis ("Alzheimer's") was observed are shown for the examples according to the present invention, but for the comparative examples, no effect was observed even at a concentration of 1 mg / ml.

[0178] Examples 42 to 82, Comparative Examples 9 to 16 - Parkinson's Test The same substance as above was tested for potential suitability as a therapeutic agent for Parkinson's disease in a 96-well procedure in a standardized α-synuclein test system using the transgenic C. elegans NL5901 strain in the first-mentioned screening assay using the nematode Caenorhabditis elegans (Pretsch et al., supra). This nematode strain expresses human α-synuclein conjugated to green fluorescent protein (GFP) in all muscle cells. With GFP labeling, the expressed protein levels can be directly detected using a multiplate reader for 24 to 48 h. In this test model, a significant decrease in the expression of neurotoxic α-synuclein should be observed with the active substance at a concentration at which the expression of α-synuclein is decreased compared to the control substance.

[0179] For this purpose, the substance was again tested at concentrations of 1 mg / ml, 100 μg / ml, and 10 μg / ml, this time compared with the first-mentioned Parkinson's disease therapeutic agent levodopa as a control. Also, the test results are shown superimposed in the following table, and the concentration ranges (μg / ml) in which a decrease in α-synuclein expression ("Parkinson's") was detected are again shown for the examples according to the present invention, but for the comparative examples of this test, no effect was again observed even at a concentration of 1 mg / ml.

[0180]

Table 1-1

[0181]

Table 1-2

[0182]

Table 1-3

[0183] The above test results clearly show that malonic acid and its multiple derivatives are promising drugs that may be effective against both Alzheimer's disease and Parkinson's disease, and even Huntington's disease and prion disease.

[0184] The results obtained with various substitution patterns show that the active molecule can include not only free carboxyl groups but also salts, esters, and substituted or unsubstituted amides without impairing its effectiveness. Since both small groups (methyl, ethyl) and larger groups (e.g., dimethyladamantyl and phenylpiperazinyl) showed equivalent effectiveness, the bulkiness of the substituent does not seem to play an important role.

[0185] As can be seen from the comparative examples, since it was proven that neither dimethylmalonic acid (101) nor dimethyl malonate dimethyl ester (102) is effective, this effectiveness is clearly based particularly on the presence of the unsubstituted malonic acid moiety. As the ineffectiveness of compounds (103) to (106) proves, the substituents contained in the effective malonic acid derivatives according to the present invention, such as (dimethyl)adamantyl groups and aminopropyl groups, have little influence on the degree of potency. The ineffectiveness of tetrahydroquinoline amide (108) is not due to the presence of aromatic substituents, of course. In particular, although non-aromatic decahydroquinoline amide (24) and its isomer decahydroisoquinoline amide (25) were already effective at concentrations exceeding 10 μg / ml, all phenylpiperazine amide derivatives were also positive.

[0186] Furthermore, the dilution rate of 1:10 selected in a series of tests leaves a wide margin for the actually effective concentration. This does not rule out the possibility that a compound that is already effective at a concentration exceeding, for example, 10 μg / ml and is thus evaluated as "++" may be effective only at a concentration just below the limit value of 100 μg / ml, for example, 90 μg / ml or higher. On the other hand, the effective limit of a compound that is effective only at a concentration exceeding 100 μg / ml and is thus evaluated as "+" may slightly exceed the limit value. The effectiveness of such a compound is actually only a few percent different.

[0187] Also, it should be borne in mind that the amount of each compound actually obtained in the test, i.e., the molar amount, naturally depends on the molecular weight, and the molecular weight varies within a wide range. For example, free malonic acid (1) (104.06 g / mol) and malonic acid diamide (2) (102.09 g / mol) have a molecular weight of 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 ratio of about 5 already predicts almost half of the dilution rate of 1:10. However, the three derivatives of malonic acid itself with relatively low molecular weights were effective only when the concentration exceeded 100 μg / ml, while the compound (41) with the second highest molecular weight after the aromatic compound (36), i.e., 460.62 g / mol, was already effective when exceeding 10 μg / ml.

[0188] Therefore, further investigation regarding the optimization of effectiveness by further varying the substitution pattern and the dilution rate in the test is currently the subject of research by the inventors.

[0189] Therefore, the present invention provides a group of compounds that should be suitable for the treatment of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease because they can induce the endogenous biosynthesis of metallothionein even at extremely low concentrations due to their effectiveness in the C. elegans assay.

Claims

1. Malonic acid of the following formula (I) and its derivatives for use as a promoter of metallothionein biosynthesis in a patient: 【Chemical Formula 53】 (I) wherein X and Y are each independently -O － M ＋ , -OR １ , and -NR ２ R ３ is selected from, M ＋ is selected from monovalent or polyvalent metal anions, R １ , R ２ and R ３ are each independently selected from hydrogen and saturated or unsaturated linear, branched or cyclic hydrocarbon radicals having from 1 to 25 carbon atoms, where one or more carbon atoms may be substituted by O or N, R ２ and R ３ may combine to form a heterocyclic hydrocarbon radical together with the nitrogen atom.

2. In formula (I), both X and Y are -O － M ＋ where each M ＋ is independently selected from monovalent and polyvalent metal anions, and M ＋ preferably represents an alkali metal or alkaline earth metal ion, more preferably represents an alkali metal ion, a compound for use according to claim 1.

3. R １ , R ２ and R ３ are each independently selected from hydrogen and saturated or unsaturated straight-chain, branched or cyclic carbon atoms having 1 to 15 carbon atoms, a compound for use according to claim 1 or 2.

4. R １ , R ２ and R ３ is each independently selected from hydrogen and saturated or unsaturated carbon atoms having from 1 to 10 carbon atoms, a compound for use according to claim 3, characterized in that.

5. A 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 diamide, disodium malonate, dimethyl malonate, potassium methyl malonate, methyl malonate amide, N-(3-aminopropyl) malonamide, adamantan-1-yl malonate, adamantan-1-yl ethyl malonate, adamantan-1-yl malonate -N-(3-aminopropyl) amide, adamantan-1-yl malonate -N-(3-aminopropyl)-N-methyl amide, adamantan-1-yl malonate -piperazine amide, adamantan-1-yl malonate -4-(4-aminobutyryl) piperazine amide, adamantan-1-yl malonate -4-aminopiperidine amide, adamantan-1-yl malonate -(4-(4-aminobutyryl)amino)piperidine amide, methyl malonate -N-(adamantan-1-yl) amide, N-(adamantan-1-yl) malonamide, N-adamantan-1-yl-N'-(3-aminopropyl) malonic diamide, N-adamantan-1-yl-N'-(3-aminopropyl)-N'-methyl malonic diamide, N-adamantan-1-yl-N'-piperazine malonic diamide, N-adamantan-1-yl-N'-(4-(4-aminobutyryl)piperazine) malonic diamide, N-(3-aminopropyl)-N'-morpholine malonic diamide, N-(3-aminopropyl)-N'-piperidine malonic diamide, N-(3-aminopropyl)-N'-decahydroquinoline malonic diamide, N-(3-aminopropyl)-N'-decahydroisoquinoline malonic diamide, N-(3-aminopropyl)-N'-(6,6-Dimethylbicyclo[3.1.1]heptan-2-ylmethyl)malonic diamide, adamantan-1-yl malonate-N-(3-dimethylaminopropyl)amide, adamantan-1-yl malonate-N-(3-morpholinopropyl)amide, adamantan-1-yl malonate-4-(3-trifluoromethylbenzyl)piperazineamide, adamantan-1-yl malonate-4-(3-fluorobenzyl)piperazineamide, adamantan-1-yl malonate-4-(2-methoxyphenyl)piperazineamide, malonic acid-N-(3,5-dimethyl)adamantan-1-ylamide, N-(3,5-dimethyladamantan-1-yl)-N'-(3-aminopropyl)malonic diamide, N-(3,5-dimethyladamantan-1-yl)-N'-(3-morpholinopropyl)malonic diamide, N-(3,5-dimethyladamantan-1-yl)-N'-piperazine malonic diamide, N-(3,5-dimethyladamantan-1-yl)-N'-4-(3-trifluoromethylbenzyl)piperazine malonic diamide, N-(3,5-dimethyladamantan-1-yl)-N'-4-(3-fluorobenzyl)piperazine malonic diamide, N-(3,5-dimethyladamantan-1-yl)-N'-4-(2-methoxyphenyl)piperazine malonic diamide, N-(3,5-dimethyladamantan-1-yl)-N'-4-(4-methoxyphenyl)piperazine malonic diamide, N-(3,5-dimethyladamantan-1-yl)-N'-4-(2,4-dimethoxyphenyl)piperazine malonic diamide, and N-(3,5-dimethyladamantan-1-yl)-N'-4-(2-morpholino-2-oxoethyl)piperazine malonic diamide.,

6. A compound for use according to any one of claims 1 to 5, characterized in that promoting metallothionein biosynthesis restores metal homeostasis and thereby serves for the treatment of neurodegenerative diseases.

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

8. A 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 body of a human or animal by promoting metallothionein biosynthesis in a patient, comprising a compound as defined in any one of claims 1 to 5, at least one pharmaceutically acceptable excipient, and optionally one or more other pharmaceutically acceptable components.

10. A pharmaceutical composition according to claim 9, characterized in that it serves for the treatment of neurodegenerative diseases.

11. A pharmaceutical composition according to claim 10, characterized in that it serves for the treatment of Alzheimer's disease or Parkinson's disease.

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