阅读说明：本技术 3,6-二取代的-2-吡啶醛肟骨架 (3, 6-disubstituted-2-pyridine aldoxime skeletons ) 是由 拉希德·巴蒂 理查德·布朗 若泽·迪亚斯 亚历克斯·马扬-因斯通 贾格迪什·耶里 于 2020-02-14 设计创作，主要内容包括：本发明涉及式(I)的化合物、或其药物学上可接受的盐之一：其中,R1、R2和-X-Y-具有特定的定义。本发明还涉及此类化合物用于治疗中的用途；及该类化合物的制备方法。(The present invention relates to a compound of formula (I), or one of its pharmaceutically acceptable salts:)

1. A compound of formula (I), or one of its pharmaceutically acceptable salts:

wherein:

r1 is H, or a linear or cyclic (preferably aromatic) C1-C7 alkoxy radical, preferably R1 is H, methoxy or benzyloxy;

-X-Y-is-CH₂-(CH₂)_n-、-C≡C-、or-X-Y-is Br and R2 is absent;

n is an integer of 0 to 5;

r2 is a group selected from alkyl, aryl, aralkyl, heteroaryl, -R3-N (R4) (R5), radical a, radical B, radical C and radical D, wherein radical a or radical B or radical C or radical D may be attached to-Y-X-via an alkyl group, preferably an ethyl group:

r3 is C1-C4 alkyl, and

r4 and R5 are identical or different and each independently represents H, a naphthyl radical, a 5-fluoroquinolin-4-yl radical, a quinolin-4-yl radical or a 8-methoxyquinolin-4-yl radical, or

R4 and R5 form together with the nitrogen atom a 4-benzyl-piperazin-1-yl radical or a 3, 7-dimethyl-2, 6-dioxy-2, 3,6, 7-tetrahydro-1H-purin-1-yl radical.

2. The compound of claim 1, wherein the compound has the following backbone 1:

wherein R1 is as defined in claim 1.

3. The compound of claim 1, wherein the compound has the following backbone 2:

wherein R1 and R2 are as defined in claim 1.

4. A compound according to claim 3, wherein R2 is selected from radical a, radical B, radical C and radical D, preferably wherein radical C or radical D is linked to-Y-X-via an alkyl group, more preferably an ethyl group:

5. the compound of claim 3, wherein R2 is alkyl, heteroaryl, aralkyl, or-R3-N (R4) (R5);

wherein R3 is C1-C4 alkyl, preferably R3 is selected from methyl, ethyl and n-propyl;

r4 is H; and is

R5 is selected from naphthyl radical, 5-fluoroquinolin-4-yl radical, quinolin-4-yl radical, or 8-methoxyquinolin-4-yl radical.

6. A compound according to claim 3, wherein R2 is-R3-N (R4) (R5), wherein R3 is C1-C4 alkyl, preferably R3 is selected from methyl, ethyl and N-propyl; and is

R4 and R5 form together with the nitrogen atom a 4-benzylpiperazin-1-yl radical or a 3, 7-dimethyl-2, 6-dioxy-2, 3,6, 7-tetrahydro-1H-purin-1-yl radical.

7. The compound of claim 1, wherein the compound has the following backbone 3:

wherein R1 and R2 are as defined in claim 1, and n is an integer from 0 to 5.

8. The compound of claim 7, wherein R2 is alkyl, aryl, aralkyl, or-R3-N (R4) (R5),

wherein R3 is C1-C4 alkyl, preferably R3 is selected from methyl, ethyl and n-propyl,

r4 is H, and

r5 is selected from naphthyl radical, 5-fluoroquinolin-4-yl radical, quinolin-4-yl radical, or 8-methoxyquinolin-4-yl radical.

9. The compound of claim 7, wherein R2 is-R3-N (R4) (R5), wherein R3 is C1-C4 alkyl, preferably R3 is selected from methyl, ethyl and N-propyl, and

r4 and R5 form together with the nitrogen atom a 4-benzylpiperazin-1-yl radical.

10. The compound of claim 1, wherein the compound has the following backbone 4:

wherein R1 and R2 are as defined in claim 1.

11. A compound according to claim 10, wherein R2 is selected from radical a, radical C and radical D, preferably wherein radical C or radical D is linked to-Y-X-through an alkyl group, more preferably an ethyl group:

12. the compound of any one of claims 1 to 11, wherein the compound is selected from:

6-bromopyridine aldoxime 2:

6- (5-phenylpent-1-yn-1-yl) pyridine aldoxime 5:

6- (5-phenylpentyl) pyridine aldoxime 7:

6- (pentadecyl-1-yn-1-yl) pyridine aldoxime 9:

6-pentadecylpyridinealdoxime 10:

6- (pyridin-3-ylethynylene) pyridylaldoxime 12:

2- ((hydroxyimino) methyl) -6- (pyridin-1-chloride-3-ylethynylene) pyridine-1-chloride 13:

n- (4- {6- [ (hydroxyimino) methyl ] pyridin-2-yl } but-3-yn-1-yl) naphthalen-1-amine 19:

n- (4- {6- [ (hydroxyimino) methyl ] pyridin-2-yl } but-3-yn-1-yl) naphthalen-1-amine 20:

6- (4- (quinolin-4-ylamino) but-1-yn-1-yl) pyridine aldoxime 25:

3-hydroxy-6- (4- (quinolin-4-ylamino) butyl) picolinic acid methyl ester 26:

6- (4- ((5-fluoroquinolin-4-yl) amino) but-1-yn-1-yl) pyridine aldoxime 30:

6- (4- ((5-fluoroquinolin-4-yl) amino) butyl) pyridine aldoxime 31:

6- (4- ((8-methoxyquinolin-4-yl) amino) but-1-yn-1-yl) pyridine aldoxime 36:

6- (4- ((8-methoxyquinolin-4-yl) amino) butyl) pyridine aldoxime 37:

6- (3- (4-benzylpiperazin-1-yl) prop-1-yn-1-yl) pyridine aldoxime 42:

6- (3- (4-benzylpiperazin-1-yl) propyl) pyridine aldoxime 43:

6- (4- (4-benzylpiperazin-1-yl) but-1-yn-1-yl) pyridylaldoxime 47:

6- (4- (4-benzylpiperazin-1-yl) butyl) pyridine aldoxime 48:

6- (4- (3, 7-dimethyl-2, 6-dioxy-2, 3,6, 7-tetrahydro-1H-purin-1-yl) but-1-yn-1-yl) pyridine aldoxime 51:

(3aS,4S,6R,6aR) -6- (6-amino-9H-purin-9-yl) -N- (4- (6- (hydroxyimino) methyl) pyridin-2-yl) but-3-yn-1-yl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxol-4-carboxamide 56:

(2S,3S,4R,5R) -5- (6-amino-9H-purin-9-yl) -3, 4-dihydroxy-N- (4- (6- (hydroxyimino) methyl) pyridin-2-yl) but-3-yn-1-yl) tetrahydrofuran-2-carboxamide 57:

n- (9- ((3aR,4R, 6aR) -6- (((3- (6- (hydroxyimino) methyl) pyridin-2-yl) prop-2-yn-1-yl) oxy) methyl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolan-4-yl) -9H-purin-6-yl) benzamide 60:

6- (3- (((3aR,4R,6R,6aR) -6- (6-amino-9H-purin-9-yl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolan-4-yl) methoxy) prop-1-yn-1-yl) pyridinaldoxime 61:

3-methoxy-6- (5-phenylpent-1-yn-1-yl) pyridine aldoxime 64:

3-methoxy-6- (5-phenylpentyl) pyridine aldoxime 65:

3-methoxy-6- (4- (quinolin-4-ylamino) but-1-yn-1-yl) pyridine aldoxime 67:

4- ((4- (6- ((hydroxyimino) methyl) -5-methoxypyridin-2-yl) but-3-yn-1-yl) amino) quinoline-1-chloride 68:

3-methoxy-6- (4- (quinolin-4-ylamino) butyl) pyridine aldoxime 69:

4- ((4- (6- ((hydroxyimino) methyl) -5-methoxypyridin-2-yl) butyl) amino) quinolin-1-quat 70:

(3aS,4S,6R,6aR) -6- (6-amino-9H-purin-9-yl) -N- (4- (6- ((hydroxyimino) methyl) -5-methoxypyridin-2-yl) but-3-yn-1-yl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolane-4-carboxamide 72:

(3aS,4S,6R,6aR) -6- (6-amino-9H-purin-9-yl) -N- (2- (1- ((6- ((hydroxyimino) methyl) pyridin-2-yl) methyl) -1H-1,2, 3-triazol-4-yl) ethyl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolane-4-carboxamide 77:

(2S,3S,4R,5R) -5- (6-amino-9H-purin-9-yl) -3, 4-dihydroxy-N- (2- (1- ((6- ((hydroxyimino) -methyl) pyridin-2-yl) methyl) -1H-1,2, 3-triazol-4-yl) ethyl) tetrahydrofuran-2-carboxamide hydrochloride 78:

n- (9- ((3aR,4R, 6aR) -6- (((1- ((6- ((hydroxyimino) methyl) pyridin-2-yl) methyl) -1H-1,2, 3-triazol-4-yl) methoxy) methyl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolan-4-yl) -9H-purin-6-yl) benzamide 79:

more preferably, the compound is selected from:

(2S,3S,4R,5R) -5- (6-amino-9H-purin-9-yl) -3, 4-dihydroxy-N- (4- (6- (hydroxyimino) methyl) pyridin-2-yl) but-3-yn-1-yl) tetrahydrofuran-2-carboxamide 57:

6- (4- (quinolin-4-ylamino) but-1-yn-1-yl) pyridine aldoxime 25:

3-hydroxy-6- (4- (quinolin-4-ylamino) butyl) picolinic acid methyl ester 26:

(2S,3S,4R,5R) -5- (6-amino-9H-purin-9-yl) -3, 4-dihydroxy-N- (2- (1- ((6- ((hydroxyimino) -methyl) pyridin-2-yl) methyl) -1H-1,2, 3-triazol-4-yl) ethyl) tetrahydrofuran-2-carboxamide hydrochloride 78:

13. process for the preparation of a compound of formula (I) according to any one of claims 1 to 12, wherein-X-Y-is-CH₂-CH₂-or-C ≡ C-, the process comprising a reduction step of a sonogashira coupling reaction between 6-bromopyridine aldoxime and a compound comprising a terminal alkyne, optionally followed by reduction by reaction with hydrogen.

14. A compound according to any one of claims 1 to 12, wherein the compound is for use in therapy.

15. The compound of any one of claims 1 to 12, wherein the compound is for use in the treatment of nerve failure and/or respiratory failure due to poisoning by at least one organophosphorous nerve agent; and/or for the treatment of neurological diseases such as alzheimer's disease or parkinson's disease; and/or for the treatment of inflammation; and/or for the treatment of cancer; and/or for the treatment of diabetes; and/or for the treatment of pain.

Technical Field

The present invention relates to a novel compound having a 3, 6-disubstituted-2-pyridinealdoxime skeleton. Such compounds are useful in a number of therapeutic and non-therapeutic applications. The invention also relates to compositions, especially pharmaceutical compositions, comprising said compounds; and uses thereof.

Background

Organophosphorus nerve agents (OPNA) are highly toxic compounds including Chemical Warfare Agents (CWA) including sarin, soman, cyclosarin, taben (Tabu), O-ethyl S- [2- (diisopropylamino) ethyl ] methyl thiophosphate (VX) and pesticides such as paraoxon, parathion and tetraethyl pyrophosphate (TEPP). Their acute toxicity is due to irreversible inhibition of acetylcholinesterase (AChE) by its catalytic phosphorylation of serine, resulting in the inability of the enzyme to hydrolyze acetylcholine (ACh). This neurotransmitter accumulates at cholinergic synapses, resulting in permanent saturation of muscarinic and nicotinic receptors, ultimately leading to seizure and cessation of breathing. Depending on the type of OPNA and the dose administered, death may occur within minutes.

Efforts to control the diffusion of these agents have proven to be limited due to the similarity between chemical weapons and the chemical precursors of pesticides and the relatively simple chemical components involved in their synthesis. Therefore, developing effective measures against OPNA poisoning remains a challenging problem for protecting and treating civilian and military personnel. Current treatment of OPNA intoxication involves the combined use of atropine (an antimycin drug) and diazepam (an anticonvulsant drug) and standard pyridine oximes (pranopdoxime or 2-PAM, trimethoxy oxime, HI-6, oxidopxime or oxidophytxime) To reactivate acetylcholinesterase. Oximes act on the OPNA-inhibited acetylcholinesterase by attacking the phosphorous atom of phosphorylated serine, thereby removing the phosphonate and restoring the catalytic activity of the enzyme. The hybrid reactivation agent compound has a pyridine oximido structure coupled to a potential ligand at the enzyme peripheral site known as a Peripheral Site Ligand (PSL). The aim was to increase the affinity of the activators for acetylcholinesterase (Mercey G. et al, Chemical Research publication 756-766,2012, volume 45, phase 5 (Mercey G. et al, Accounts of Chemical Research 756-766,2012, Vol.45, No. 5)).

The efficiency of reactivation can be estimated by the secondary rate constant for reactivation of kr2, which is the maximum reactivation rate constant (kr) with reactivation of the acetylcholinesterase complex inhibited (K)_D) The apparent dissociation constant of (c). To date, none of the oxime compounds known has been equally effective against all of the OPNA-inhibited acetylcholinesterases.

More recently, WO2017/021319 discloses bifunctional compounds comprising specific Peripheral Site Ligand (PSL) moieties of aminoquinoline functional groups, which increase affinity for toxic hAChE (and therefore have a lower KD), making them effective reactivators of human acetylcholinesterase for inhibition by any type of organophosphorus compounds. However, these bifunctional compounds contain a hydroxyl group, which may be present at position 3 of the pyridine radical. This hydroxyl group needs to be protected and deprotected during the synthesis. Furthermore, the hydroxyl group may participate in the internal cyclization of the molecule.

Thus, there is still a need for compounds that are effective in therapeutic applications, in particular against opa poisoning, that are fast and easy to synthesize, have good yields, and are produced on a higher scale. These compounds must be stable without any internal cyclisation.

Surprisingly, the present inventors have now found that specific pyridylaldoxime compounds (bearing a hydrogen or a specific alkoxy group at the 3-position) meet these requirements. They can readily cross the blood brain barrier, especially since they are uncharged.

In fact, such compounds are fast, simple and very easy to produce. The obtained compounds do not show intramolecular cyclization and are useful in human therapy.

Notably, these compounds can act as antidotes to OPNA poisoning, or as antidotes to organophosphorous compounds, because they effectively and rapidly reactivate hAChE. Without being bound by any theory, these molecules appear to bind selectively to the catalytic site of hAChE. They show in particular a very high reactivation efficiency of the inhibited acetylcholinesterase. Once the oxime of the compound dephosphorizes the serine residue, it can be regenerated: thus, the compound may be used multiple times. These compounds are also agonists of the adenosine 2A receptor. Thus, they are useful in the treatment of inflammation; for the treatment of neurodegenerative diseases such as alzheimer's disease or parkinson's disease; for the treatment of cancer, in particular due to the inhibitory activity of Histone Deacetylase (HDAC) in the treatment of diabetes and/or in the treatment of pain.

Accordingly, a first object of the present invention is a compound of formula (I):

wherein the different groups are defined as in the detailed description below.

Another object of the present invention is a process for the preparation of the compound of formula (I), in particular by Sonogashira coupling reaction (Sonogashira reaction), as described below.

Another object of the present invention is a pharmaceutical composition comprising at least one compound of formula (I) and at least one pharmaceutically acceptable carrier.

Another object of the present invention is a compound according to the invention for use as a medicament.

Another object of the present invention is a compound according to the invention for use in the treatment of neurological and/or respiratory failure caused by poisoning with at least one organophosphorus nerve agent.

Another object of the invention is a compound according to the invention for use in the treatment of inflammation.

Another object of the present invention are compounds according to the invention for use in the treatment of neurological diseases such as alzheimer's disease or parkinson's disease.

Another object of the present invention is a compound according to the invention for use in the treatment of cancer.

Another object of the present invention are the compounds according to the invention for use in the treatment of diabetes.

Another object of the invention is a compound according to the invention for use in the treatment of pain.

A first object of the present invention is a compound of formula (I), or one of its pharmaceutically acceptable salts:

wherein:

r1 is H, or a linear or cyclic (preferably aromatic) C1-C7 alkoxy radical, preferably R1 is methoxy or benzyloxy;

-X-Y-is-CH₂-(CH₂)_n-、-C≡C-、or-X-Y-is Br and R2 is absent;

n is an integer of 0 to 5;

r2 is a group selected from alkyl, aryl, aralkyl, heteroaryl, -R3-N (R4) (R5), radical a, radical B, radical C and radical D, wherein radical a or radical B or radical C or radical D may be attached to-Y-X-via an alkyl group, preferably an ethyl group:

r3 is C1-C4 alkyl, and

r4 and R5 are the same or different and each independently represents H, a naphthyl radical, a 5-fluoroquinolin-4-yl radical, a quinolin-4-yl radical or an 8-methoxyquinolin-4-yl radical, or

R4 and R5 form together with the nitrogen atom a 4-benzylpiperazin-1-yl radical or a 3, 7-dimethyl-2, 6-dioxy-2, 3,6, 7-tetrahydro-1H-purin-1-yl radical.

For the definition of-X-Y-, the point of attachment of the triazole group is indicated by an asterisk on each side of the triazole group:

the point of attachment of any one of the moieties a to D (in the definition of R2) to the remainder of the molecule of formula (I) is indicated by an asterisk:

"Bz" in radical a denotes benzoyl, i.e. Ph-C (═ O) -.

"pharmaceutically acceptable salt" refers to any salt of a compound of formula (I) with an acid or base. The pharmaceutically acceptable salt may be a hydrochloride salt. The salts may be obtained using a pyridine of formula (I) to give pyridine salts. For example, when R4 and/or R5 are the same or different and each independently represents a 5-fluoroquinolin-4-yl radical, a quinolin-4-yl radical, or an 8-methoxyquinolin-4-yl radical, the radicals may be complexed with HCl to give a 5-fluoroquinolin-4-yl radical, a 4-quinolinyl radical, or an 8-methoxy-4-quinolinyl radical, respectively. Preferred pharmaceutically acceptable salts are the 5-fluoro-4-quinoline, 4-quinoline and 8-methoxy-4-quinoline radicals.

The oximes of the compounds of formula (I) may be labelled with one or more isotopes, for example,¹⁵N、¹⁸O、²h or³H. In fact, such stable, non-toxic, non-radioactive isotopes would allow biological studies in vivo and in vitro.

"alkyl" means a linear hydrocarbon radical preferably containing from 1 to 20 carbon atoms, in particular from 1 to 15 carbon atoms, or a branched or cyclic hydrocarbon radical containing from 3to 20 carbon atoms. Examples of the alkyl group include a methyl group, an ethyl group, a n-propyl group, an isopropyl group, a n-butyl group, an isobutyl group, a tert-butyl group, a n-pentyl group, an isopentyl group, a n-hexyl group, a n-tridecyl group, a cyclohexyl group and a cyclohexylmethyl group, and preferably an ethyl group, a propyl group, a n-hexyl group, a n-tridecyl group, a cyclohexyl group or a cyclohexylmethyl group.

C1-C4 alkyl is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl.

"Linear C1-C7 alkoxy radical" refers to the radical Rad-O-, where Rad is a linear C1-C7 alkyl group. Preferably, the linear C1-C7 alkoxy radical is methoxy. By "cyclic (preferably aromatic) C1-C7 alkoxy radical" is meant the radical Rad-O-where Rad is a cyclic, preferably aromatic, C1-C7 alkyl group. Preferably, the cyclic C1-C7 alkoxy radical is an aromatic C1-C7 alkoxy radical, and more preferably benzyloxy.

"aryl" refers to a monocyclic or polycyclic aromatic hydrocarbon group which may be optionally substituted. Preferably, the aryl group is phenyl or Polycyclic Aromatic Hydrocarbon (PAH). The preferred polycyclic aromatic hydrocarbon is pyrene. The aryl group may be substituted with at least one alkyl group and/or with at least one cyano (-CN). A preferred example of aryl is phenyl.

"aralkyl" refers to an aryl group as described above which is attached to the compound of formula (I) through an alkyl group. Preferably, aralkyl is phenylpropyl. An aralkyl group may be substituted on the aryl group with at least one alkyl group and/or with at least one cyano group (-CN). A preferred aralkyl group is phenylpropyl.

"heteroaryl" refers to an aryl group in which at least one carbon atom of the aromatic ring is substituted with a heteroatom, and may be optionally substituted. The heteroatom may be nitrogen, oxygen, phosphorus or sulfur. Preferably, the heteroatom is nitrogen. Examples of heteroaryl groups include pyrrole, thiophene, furan, pyridine, pyrimidine, pyrazine, triazine, imidazole, thiazole, oxazole and isoxazole groups. Preferably, heteroaryl is pyridyl, e.g., 4-or 3-pyridyl. The heteroaryl group may be substituted with at least one alkyl group and/or at least one cyano (-CN). Preferably, the heteroaryl group is in the form of a salt, preferably a pyridyl group, e.g., 4-or 3-pyridine.

According to a first embodiment, preferably in formula (I) -X-Y-is Br and R2 is absent.

Thus, a compound of formula (I) or one of its pharmaceutically acceptable salts has the following backbone 1:

skeleton 1

Wherein R1 is as defined above.

The compound of backbone 1 is a compound wherein R1 is H or a linear or cyclic (preferably aromatic) C1-C7 alkoxy radical. Preferably, the compound of backbone 1 is a compound wherein R1 is H, methoxy or benzyloxy.

According to a second embodiment, preferably-X-Y-in formula (I) is-C-Y- (backbone 2):

skeleton 2

Wherein R1 and R2 are as defined above.

The compound of backbone 2 is a bifunctional compound.

Preferably, the compound of backbone 2 is a compound wherein R1 is selected from H and methoxy.

Preferably, the compound of backbone 2 is a compound wherein R2 is selected from the group consisting of radical a, radical B, radical C and radical D, preferably wherein radical C or radical D is linked to-Y-X-via an alkyl group, more preferably an ethyl group:

alternatively, preferably, the compound of backbone 2 is a compound wherein R2 is alkyl, heteroaryl, aralkyl or-R3-N (R4) (R5),

wherein R3 is C1-C4 alkyl, preferably R3 is selected from methyl, ethyl and n-propyl,

r4 is H, and

r5 is selected from naphthyl radical, 5-fluoroquinolin-4-yl radical, quinolin-4-yl radical, or 8-methoxyquinolin-4-yl radical.

Alternatively, preferably, the compound of backbone 2 is a compound wherein R2 is-R3-N (R4) (R5) wherein R3 is C1-C4 alkyl, preferably R3 is selected from methyl, ethyl and N-propyl, and

r4 and R5 form together with the nitrogen atom a 4-benzylpiperazin-1-yl radical or a 3, 7-dimethyl-2, 6-dioxy-2, 3,6, 7-tetrahydro-1H-purin-1-yl radical.

Compounds of framework 2 have a reduced pKa and increased reactivation efficiency due to increased affinity compared to compounds without triple binding and with an-OH group of R1 and compared to reference molecules such as pralidoxime (2-PAM) and HI-6 (kr2 units: mM)^-1min^-1)。

According to a third embodiment, in formula (I) preferably-X-Y-is-CH₂-(CH₂)_n-, where n is an integer of 0 to 5 (backbone 3):

skeleton 3

Wherein R1 and R2 are as defined above.

The compound of backbone 3 is a bifunctional compound.

Preferably, the compound of backbone 3 is a compound wherein R1 is selected from H and methoxy.

Preferably, the compound of backbone 3 is a compound wherein R2 is alkyl, aryl, aralkyl or-R3-N (R4) (R5),

wherein R3 is C1-C4 alkyl, preferably R3 is selected from methyl, ethyl and n-propyl,

r4 is H, and

r5 is selected from naphthyl radical, 5-fluoroquinolin-4-yl radical, quinolin-4-yl radical, or 8-methoxyquinolin-4-yl radical.

Alternatively, preferably, the compound of backbone 3 is a compound wherein R2 is-R3-N (R4) (R5) wherein R3 is C1-C4 alkyl, preferably R3 is selected from methyl, ethyl and N-propyl, and

r4 and R5 form together with the nitrogen atom a 4-benzylpiperazin-1-yl radical.

According to a fourth embodiment, in formula (I) preferably-X-Y-is(backbone 4):

skeleton 4

Wherein R1 and R2 are as defined above.

The compound of backbone 4 is a trifunctional compound.

Preferably, the compound of backbone 4 is a compound wherein R1 is selected from H, methoxy and benzyloxy; preferably, R1 is H.

Preferably, the compound of backbone 4 is a compound wherein R2 is selected from the group consisting of radical a, radical C and radical D, preferably wherein radical C or radical D is linked to-Y-X-via an alkyl group, more preferably an ethyl group:

the compounds of backbone 4 selectively target the catalytic site of hAChE and show good reactivation kinetics.

Preferably, the compound of formula (I) is selected from the following compounds and pharmaceutically acceptable salts thereof:

6-bromopyridine aldoxime 2:

6- (5-phenylpent-1-yn-1-yl) pyridine aldoxime 5:

6- (5-phenylpentyl) pyridine aldoxime 7:

6- (pentadecyl-1-yn-1-yl) pyridine aldoxime 9:

6-pentadecylpyridinealdoxime 10:

6- (pyridin-3-ylethynylene) pyridylaldoxime 12:

2- ((hydroxyimino) methyl) -6- (pyridin-1-chloride-3-ylethynylene) pyridine-1-chloride 13:

n- (4- {6- [ (hydroxyimino) methyl ] pyridin-2-yl } but-3-yn-1-yl) naphthalen-1-amine 19:

n- (4- {6- [ (hydroxyimino) methyl ] pyridin-2-yl } but-3-yn-1-yl) naphthalen-1-amine 20:

6- (4- (quinolin-4-ylamino) but-1-yn-1-yl) pyridine aldoxime 25:

3-hydroxy-6- (4- (quinolin-4-ylamino) butyl) picolinic acid methyl ester 26:

6- (4- ((5-fluoroquinolin-4-yl) amino) but-1-yn-1-yl) pyridine aldoxime 30:

6- (4- ((5-fluoroquinolin-4-yl) amino) butyl) pyridine aldoxime 31:

6- (4- ((8-methoxyquinolin-4-yl) amino) but-1-yn-1-yl) pyridine aldoxime 36:

6- (4- ((8-methoxyquinolin-4-yl) amino) butyl) pyridine aldoxime 37:

6- (3- (4-benzylpiperazin-1-yl) prop-1-yn-1-yl) pyridine aldoxime 42:

6- (3- (4-benzylpiperazin-1-yl) propyl) pyridine aldoxime 43:

6- (4- (4-benzylpiperazin-1-yl) but-1-yn-1-yl) pyridylaldoxime 47:

6- (4- (4-benzylpiperazin-1-yl) butyl) pyridine aldoxime 48:

6- (4- (3, 7-dimethyl-2, 6-dioxy-2, 3,6, 7-tetrahydro-1H-purin-1-yl) but-1-yn-1-yl) pyridine aldoxime 51:

(3aS,4S,6R,6aR) -6- (6-amino-9H-purin-9-yl) -N- (4- (6- (hydroxyimino) methyl) pyridin-2-yl) but-3-yn-1-yl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxol-4-carboxamide 56:

(2S,3S,4R,5R) -5- (6-amino-9H-purin-9-yl) -3, 4-dihydroxy-N- (4- (6- (hydroxyimino) methyl) pyridin-2-yl) but-3-yn-1-yl) tetrahydrofuran-2-carboxamide 57:

n- (9- ((3aR,4R, 6aR) -6- (((3- (6- (hydroxyimino) methyl) pyridin-2-yl) prop-2-yn-1-yl) oxy) methyl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolan-4-yl) -9H-purin-6-yl) benzamide 60:

6- (3- (((3aR,4R,6R,6aR) -6- (6-amino-9H-purin-9-yl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolan-4-yl) methoxy) prop-1-yn-1-yl) pyridinaldoxime 61:

3-methoxy-6- (5-phenylpent-1-yn-1-yl) pyridine aldoxime 64:

3-methoxy-6- (5-phenylpentyl) pyridine aldoxime 65:

3-methoxy-6- (4- (quinolin-4-ylamino) but-1-yn-1-yl) pyridine aldoxime 67:

4- ((4- (6- ((hydroxyimino) methyl) -5-methoxypyridin-2-yl) but-3-yn-1-yl) amino) quinoline-1-chloride 68:

3-methoxy-6- (4- (quinolin-4-ylamino) butyl) pyridine aldoxime 69:

4- ((4- (6- ((hydroxyimino) methyl) -5-methoxypyridin-2-yl) butyl) amino) quinolin-1-quat 70:

(3aS,4S,6R,6aR) -6- (6-amino-9H-purin-9-yl) -N- (4- (6- ((hydroxyimino) methyl) -5-methoxypyridin-2-yl) but-3-yn-1-yl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolane-4-carboxamide 72:

(3aS,4S,6R,6aR) -6- (6-amino-9H-purin-9-yl) -N- (2- (1- ((6- ((hydroxyimino) methyl) pyridin-2-yl) methyl) -1H-1,2, 3-triazol-4-yl) ethyl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolane-4-carboxamide 77:

(2S,3S,4R,5R) -5- (6-amino-9H-purin-9-yl) -3, 4-dihydroxy-N- (2- (1- ((6- ((hydroxyimino) -methyl) pyridin-2-yl) methyl) -1H-1,2, 3-triazol-4-yl) ethyl) tetrahydrofuran-2-carboxamide hydrochloride 78:

n- (9- ((3aR,4R, 6aR) -6- (((1- ((6- ((hydroxyimino) methyl) pyridin-2-yl) methyl) -1H-1,2, 3-triazol-4-yl) methoxy) methyl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolan-4-yl) -9H-purin-6-yl) benzamide 79:

more preferably, the compound of formula (I) is selected from the following compounds:

(2S,3S,4R,5R) -5- (6-amino-9H-purin-9-yl) -3, 4-dihydroxy-N- (4- (6- (hydroxyimino) methyl) pyridin-2-yl) but-3-yn-1-yl) tetrahydrofuran-2-carboxamide 57:

6- (4- (quinolin-4-ylamino) but-1-yn-1-yl) pyridine aldoxime 25:

3-hydroxy-6- (4- (quinolin-4-ylamino) butyl) picolinic acid methyl ester 26:

(2S,3S,4R,5R) -5- (6-amino-9H-purin-9-yl) -3, 4-dihydroxy-N- (2- (1- ((6- ((hydroxyimino) -methyl) pyridin-2-yl) methyl) -1H-1,2, 3-triazol-4-yl) ethyl) tetrahydrofuran-2-carboxamide hydrochloride 78:

preparation of Compounds of formula (I)

The compound of formula (I) according to the invention or one of its pharmaceutically acceptable salts can be synthesized by any suitable method. For example, a compound of formula (I) or one of its pharmaceutically acceptable salts may be prepared according to the following scheme:

reacting a compound of formula (I) of backbone 1 (i.e. wherein X-Y is Br) with R2-X-Y-H (wherein X-Y is-C.ident.C-) to obtain a compound of formula (I) of backbone 2 (wherein X-Y is-C.ident.C-).

Then, by selective hydrogenation (by H)₂) Compounds of formula (I) wherein X-Y is-CH are readily available₂-CH₂- (framework 3).

To obtain the compound of backbone 4, alkyne R2-C ≡ CH is reacted with the corresponding linkable reactivator by linking chemistry.

The following examples illustrate these methods.

Preparation of Compounds of formula (I)

The compound of formula (I) according to the invention or one of its pharmaceutically acceptable salts can be synthesized by any suitable method known to the person skilled in the art.

Preferably, the compounds of formula (I) are synthesized according to the methods described below. This method is chemoselective. In particular, it does not require any prior oxime protection steps. The method comprises a minimum number of steps (one or two) and is performed rapidly at ambient temperature.

Skeleton 1

Specifically, compounds of backbone 1:

skeleton 1

Can be obtained by reacting a pyridylaldehyde precursor or a pyridonitrile derivative with hydroxylamine hydrochloride, preferably in an organic solvent. In all cases, hydroxylamine hydrochloride can be used¹⁵And marking the N element.

Such synthesis of 6-bromopyridine aldoxime 2 is illustrated in the examples.

Skeleton 2

In particular, the process for the synthesis of the compound of formula (I) (i.e. backbone (2)) may comprise the late step of the sonogashira coupling reaction between the compound of backbone 1 (i.e. 6-bromopyridine aldoxime) and the compound containing a terminal alkyne (see figure above), preferably consisting of the late step of the sonogashira coupling reaction between the compound of backbone 1 (i.e. 6-bromopyridine aldoxime) and the compound containing a terminal alkyne.

Such sonogashira coupling reactions can be carried out in the presence of a compound such as Tetrahydrofuran (THF), triethylamine (Et)₃N) or preferably mixtures thereof; in catalysts such as Pd (PPh)₃)₄And CuI.

This sonogashira coupling reaction is carried out without protecting the oxime moiety.

Skeleton 3

The alkyne obtained (backbone 2) can then be reduced by reaction with hydrogen, for example in the presence of a palladium catalyst (e.g. Pd/C), to obtain the corresponding alkyl group (backbone 3) in a selective hydrogenation step.

Also, the hydrogenation step is carried out without protecting the oxime moiety.

Skeleton 4

As described above and illustrated in the above schemes, to obtain compounds of backbone 4, the alkyne R2-C ≡ CH is reacted with the corresponding linkable reactivator by linking chemistry.

Accordingly, an object of the present invention is a process for the preparation of compounds of formula (I), wherein-X-Y-is-CH₂-CH₂-or-C ≡ C-, R1 and R2 are as defined above, including the step of sonogashira coupling reaction between 6-bromopyridine aldoxime and a compound containing a terminal alkyne, optionally followed by a reduction reaction by reaction with hydrogen.

Medical use of the compounds of the invention

The compounds of the invention may be used for the treatment of neurological and/or respiratory failure due to poisoning by at least one organophosphorus nerve agent, which may preferably be selected from warfare agents such as VX, taben (Tabun), sarin, cyclosarin and soman, and pesticides such as paraoxon, parathion and TEPP. The compounds of the invention are useful in the treatment of neurological and/or respiratory failure due to poisoning by at least one organophosphorus nerve agent, because they have a potent organophosphorus reactivation effect against cholinesterase.

These compounds may alternatively be used for the treatment of diseases involving a reduction in acetylcholine production, which may be overcome by administration of acetylcholinesterase inhibitors. Examples of such diseases include, in particular, neurological diseases such as Alzheimer's disease or Parkinson's disease.

The compounds of the present invention are also agonists of the adenosine 2A receptor. Thus, they are useful in the treatment of inflammation; for the treatment of cancer; for the treatment of diabetes; and/or for the treatment of pain.

The compounds of the present invention are typically included in a pharmaceutical composition comprising at least one compound of the present invention and a pharmaceutically acceptable carrier.

The amount of a compound of formula (I) or one of its pharmaceutically acceptable salts in the composition according to the invention may vary within a wide range depending on the patient, the mode of administration and the desired effect.

The compounds or compositions according to the invention may be administered orally or parenterally, for example, by the topical, parenteral, intramuscular, intravenous, cutaneous, nasal or rectal route.

The pharmaceutical compositions of the present invention may take various forms including granules, powders, tablets, capsules, syrups, emulsions, suspensions and forms for non-oral administration, for example, injections, sprays, transdermal patches or suppositories. These pharmaceutical forms can be prepared by known conventional techniques.

For example, the preparation of solid pharmaceutical forms for oral administration can be carried out by the following procedure: excipients (e.g., lactose, sucrose, starch, or mannitol), dehydrating agents (e.g., calcium carbonate, calcium carboxymethylcellulose, alginic acid, sodium carboxymethylcellulose, colloidal silicon dioxide, croscarmellose sodium, crospovidone, guar gum, magnesium aluminum silicate, microcrystalline cellulose, powdered cellulose, pregelatinized starch, sodium alginate, or glycolstarch ester), a binder (e.g., α -starch, gum arabic, carboxymethyl cellulose, polyvinyl pyrrolidone, hydroxypropyl cellulose, alginic acid, carbomer, dextrin, ethyl cellulose, sodium alginate, maltodextrin, liquid glucose, magnesium aluminum silicate, hydroxyethyl cellulose, methyl cellulose, or guar gum) and a lubricant (e.g., talc, magnesium stearate, or polyethylene 6000) are added to the active compound, and the resulting mixture is then tableted. If necessary, the tablets may be coated by known techniques to mask the taste (e.g. with cocoa, mint, borneol or cinnamon powder) or to allow enteric dissolution or sustained release of the active compound. Among the coating products that may be used are: for example, ethyl cellulose, hydroxymethyl cellulose, polyoxyethylene glycol, cellulose acetate phthalate, hydroxypropylmethyl cellulose phthalate and(methacrylic-acrylic acid copolymer),(hydroxypropylmethylcellulose + polyethylene glycol + titanium oxide + lactose monohydrate). A pharmaceutically acceptable colorant (e.g., yellow iron oxide, red iron oxide, or quinoline yellow) may be added.

Liquid pharmaceutical forms for oral administration include solutions, suspensions and emulsions. Aqueous solutions may be obtained by dissolving the active ingredient in water, followed by the addition of flavorings, colorants, stabilizers, and/or thickeners, if necessary. To improve solubility, ethanol, propylene glycol or any other pharmaceutically acceptable non-aqueous solvent may be added. Aqueous suspensions for oral administration can be obtained by dispersing the finely divided active ingredient in water using viscous products, for example, natural or synthetic gums or resins, methylcellulose or sodium carboxymethylcellulose.

For example, the pharmaceutical form for injection can be obtained by the following process. A dispersant is used (for example,80、60(Nikko Chemicals), polyethylene glycol, carboxymethylcellulose or sodium alginate), preservatives (e.g. methylparaben, propylparaben, benzyl alcohol, chlorobutanol or phenol), isotonicity agents (e.g. sodium chloride, glycerol, sorbitol or glucose) and optionally other additives such as solubilizers (e.g. sodium salicylate or sodium acetate) or stabilizers (e.g. human serum albumin), the active compounds being dissolved, suspended or emulsified in an aqueous medium (e.g. distilled water, physiological saline or ringer's solution) or an oil medium (e.g. olive oil, sesame oil, cottonseed oil, corn oil or propylene glycol).

Topical (epidermal) pharmaceutical forms can be obtained from solid, semi-solid or liquid compositions containing the active compound. For example, to obtain a solid form, the active compound may be treated with excipients (e.g. lactose, mannitol, starch, microcrystalline cellulose or sucrose) and thickeners (e.g. natural gums, cellulose derivatives or acrylic polymers) in order to convert it into a powder. As mentioned above, the liquid pharmaceutical composition is prepared in substantially the same manner as the injectable dosage form. The semi-solid pharmaceutical dosage form is preferably in the form of an aqueous or oily gel or jam. These compositions may optionally comprise a pH adjusting agent (e.g., carbonic acid, phosphoric acid, citric acid, hydrochloric acid, or sodium hydroxide) and a preservative (e.g., parabens, chlorobutanol, or benzalkonium chloride).

Also described herein is a method for treating neurological and/or respiratory failure caused by poisoning with at least one organophosphorus nerve agent comprising administering at least one compound according to the present invention.

Also described herein is a method for treating a neurological disease such as alzheimer's disease or parkinson's disease comprising administering at least one compound according to the invention.

Also described herein is a method for treating inflammation comprising administering at least one compound according to the invention.

Also described herein is a method for treating cancer comprising administering at least one compound according to the invention.

Also described herein is a method for treating diabetes comprising administering at least one compound according to the invention.

Also described herein is a method for treating pain comprising administering at least one compound according to the invention.

In the context of the present invention, the term "treatment" means therapeutic, symptomatic and/or prophylactic treatment. In particular, it may refer to slowing the progression of the disease, reducing or inhibiting at least one symptom or complication, or in any way improving the health of the patient.

Administration of a compound or composition according to the invention can be performed before, during, or after exposure of the subject to the organophosphorous nerve agent.

In the present invention, the terms "subject" and "patient" are used interchangeably and refer to a human subject.

The amount of a compound of formula (I) or one of its pharmaceutically acceptable salts to be administered according to the invention may vary within a wide range depending on the patient, the mode of administration and the desired effect. In particular, the amount of the compound of formula (I) or one of its pharmaceutically acceptable salts may be comprised between 200mg and 4000mg, up to a dose of 3 times per day.

The compound or composition according to the invention may be co-administered with at least one other active agent, for example, an antimycin agent (especially atropine), an anticonvulsant agent (especially diazepam or a prodrug thereof, e.g. abamectin) and/or a biological scavenger capable of capturing and/or degrading OPNA in the blood, such as human butyrylcholinesterase.

The term "co-administration" means that administration of a compound or composition according to the invention and administration of the other active agent may be simultaneous, sequential and/or separate.

Other uses of the Compounds of the invention

The compounds of the invention may further be used as tools for biological studies in vivo and/or in vitro. In the present application, a compound of formula (I) or one of its pharmaceutically acceptable salts may comprise one or more isotopes which will allow its detection.

The following examples are provided as illustrative and not limiting descriptions of the present invention.

Examples

Example 1: synthesis of Compounds of the invention

Synthesis of I-bifunctional pyridoxal oxime analogs

Synthesis of 6- (5-phenylpentyl) pyridine aldoxime:

scheme 1: synthesis of 6-bromopyridine aldoxime

Scheme 2: synthesis of 6-substituted pyridine aldoximes

Scheme 3: alternative synthetic routes to 6-substituted pyridine aldoximes

Scheme 4: selective hydrogenation

6-bromopyridine aldoxime 2:

the compound is according to L.Zhang et al¹Synthesized by published work; hydroxylamine hydrochloride (2.24g, 32.3mmol) and sodium acetate (2.65g, 32.3mmol) were added to a solution of 6-bromopyridylaldehyde 1(3.00g, 16.1mmol) in anhydrous ethanol (50mL) at room temperature (rt). After the addition, the colorless solution with white suspension was stirred at 90 ℃ for 3 hours. The solution was cooled to room temperature and concentrated in vacuo. The white solid obtained was dissolved in EtOAc (50 mL). By H₂The organic layer was O washed (5X 20mL), dried (MgSO)₄) Filtered and concentrated in vacuo to afford the title compound 2(3.21g, 16.0mmol, 99%) as a white solid. The physical and spectral data were consistent with the reported values.¹mp 168-²164 ℃) and 166 ℃); IR (near Infrared) V_max 3203，3084，2912，1546，1158，1119，704cm^-1；¹HNMR(400MHz，CDCl₃)δ11.90(s，1H，CHNOH)，8.04(s，1H，CHNOH)，7.82-7.74(m，2H，NCCHCHCH，NCCHCH)，7.63(dd，J＝6.8，1.7Hz，1H，NCCHCH)；¹³C NMR(100MHz，CDCl₃)δ153.3，147.5，141.0，140.1，128.1，119.3；C₆H₅BrN₂O⁺HRMS (ESI)⁺The m/z calculation was 200.9658, and the value was found to be 200.9657.

Reference documents:

1.Bioorg.Med.Chem.Lett.2016,26,778–781

6- (5-phenylpent-1-yn-1-yl) pyridylaldehyde 4:

to THF/Et₃To a degassed solution of bromopicolinafaldehyde 1(568mg, 3.056mmol, 1.1 equiv.) in N (10mL/30mL) was added Pd [ PPh ]₃]₄(482mg, 0.0.417mmol, 0.15 equiv.) and CuI (159mg, 0.834mmol, 0.3 equiv.). After degassing the reaction mixture at room temperature for 5 minutes, alkyne 3(400mg, 2.78mmol, 1 eq.) was added dropwise and the reaction mixture was stirred at room temperature for 16 hours. Upon completion (monitored by TLC), the reaction mixture was concentrated under reduced pressure and the residue was purified by column chromatography (EtOAc/PE6:94 to EtOAc/PE 1:9) to afford the desired coupled pyridylaldehyde 4(500mg, 72%) as a colorless oil. R_f(20% EtOAc + PE) 0.65; IR (near Infrared) V_max 3026，2928，2856，2229，1710，1580，1451，1211，987，805，698，647，542cm^-1；¹HNMR(400MHz，CDCl₃)δ(ppm)9.99(s，1H，H₁₈)，7.82-7.71(m，2H，H₃，H₄)，7.53(dd，J＝7.5Hz，1H，H₅)，7.26-7.10(m，5H，H₁₃-H₁₇)，2.74(t，J＝7.5Hz，2H，H₁₁)，2.43(t，J＝7.1Hz，2H，H9)，1.92(quintet，J＝7.1，7.5Hz，2H，H₁₀)；¹³C NMR(100MHz，CDCl₃)δ(ppm)193.09(C18)，152.76(C2)，144.43(C6)，141.21(C12)，137.21(C4)，130.92(C5)，128.47(C14,C16)，128.41(C13,C17)，126.02(C15)，119.94(C3)，92.32(C7)，79.93(C8)，34.90(C11)，29.74(C10)，18.81(C9)；C₁₇H₁₆NO⁺HRMS (ESI)⁺) The m/z calculation was 250.1226, and the value was found to be 250.1239.

6- (5-phenylpent-1-yn-1-yl) pyridine aldoxime 5:

the method comprises the following steps:

acetaldehyde 4(100mg, 0.402mmol, 1 equiv.), hydroxylamine hydrochloride (56mg, 0.803mmol, 2 equiv.) and CH₃CO₂A solution of Na (100mg, 1.206mmol, 3 equiv.) in dry ethanol (6mL) was stirred at reflux for 16 h. Go toAfter completion (monitored by TLC), the solid was removed by filtration through a short pad of celite, the solvent was evaporated and the residue was purified by column chromatography (EtOAc/PE 1:9) to provide oxime 5(100mg, 94%) as a white solid. R_f(20% EtOAc + PE) 0.35; IR (near Infrared) V_max 3177，3005，2933，2876，2226，1568，1495，1445，1257，1159，985，807，734，703，657，576，490cm^-1；¹HNMR(400MHz，CDCl₃)δ(ppm)8.85(s，1H，OH)，8.24(s，1H，H₁₈)，7.68(dd，J＝0.7，7.8Hz，1H，H₃)，7.56(t，J＝7.8Hz，1H，H₄)，7.29(dd，J＝0.7，7.7Hz，1H，H₅)，7.29-7.08(m，5H，H₁₃-H₁₇)，2.71(t，J＝7.5Hz，2H，H₁₁)，2.39(t，J＝7.1H，2H，H9)，1.89(quintet,J＝7.1,7.5Hz,2H,H₁₀)；¹³C NMR(100MHz，CDCl₃)δ(ppm)151.95(C2)，150.51(C18)，144.46(C6)，141.33(C12)，136.73(C4)，128.50(C14,C16)，128.35(C13,C17)，127.13(C5)，125.92(C15)，119.23(C3)，91.47(C7)，80.19(C8)，34.84(C11)，29.78(C10)，18.77(C9)；C₁₇H₁₇N₂O₁ ⁺HRMS (ESI)⁺) The m/z calculation was 265.1335, and the value was found to be 265.1360.

The method 2 comprises the following steps:

to THF/Et₃To a degassed solution of oxime 2(77mg, 0.381mmol, 1.1 equiv) in N (5mL/2mL) was added Pd [ PPh ]₃]₄(60mg, 0.052mmol, 0.15 equiv.) and CuI (20mg, 0.104mmol, 0.3 equiv.). After degassing the reaction mixture at room temperature for 5 minutes, alkyne 3(50mg, 0.347mmol, 1 eq) was added dropwise and the reaction mixture was stirred at room temperature for 16 hours. Upon completion (monitored by TLC), the reaction mixture was concentrated under reduced pressure and the residue was purified by column chromatography (EtOAc/PE 1:9) to afford the desired coupled oxime 5(68mg, 74%) as a white solid.

6- (5-phenylpentyl) pyridylaldehyde 6:

to a degassed solution of 6-substituted pyridylaldehyde 4(200mg, 0.802mmol, 1 eq) in dry EtOAc (4mL) was added 10% Pd/C (21mg, 0.201mmol, 0.25 eq). By H₂After three washes, at room temperature and at H₂(1atm.) the reaction mixture was stirred for 90 minutes. Upon completion (monitored by TLC), the catalyst was removed by short column celite filtration, the solvent was evaporated, and the residue was purified by column chromatography (EtOAc/PE 1:9) to afford oxime 6(185mg, 91%) as a colourless liquid; r_f(20% EtOAc + PE) 0.70; IR (near Infrared) V_max 3026，2929，2856，1709，1591，1455，1213，1089，745，689，646，570，496cm^-1；¹HNMR(400MHz，CDCl₃)δ(ppm)9.97(s，1H，H₁₈)，7.73-7.63(m，2H，H₃，H₄)，7.26(dd，J＝1.5，7.7Hz，1H，H₅)，7.24-7.05(m，5H，H₁₃-H₁₇)，2.80(t，J＝7.7Hz，2H，H7)，2.54(t,J＝7.7Hz,2H,H11)，1.73(quintet，J＝7.7Hz，2H，H8)，1.60(quintet,J＝7.7Hz,2H,H₁₀)，1.35(quintet,J＝7.3,7.8Hz,2H,H₉)；¹³C NMR(100mhz，CDCl₃)δ(ppm)193.87(C18)，163.12(C6)，152.35(C2)，142.50(C12)，137.08(C4)，128.33(C14,C16)，128.20(C13,C17)，127.04(C5)，125.60(C15)，119.08(C3)，37.99(C7)，35.74(C11)，31.20(C10)，29.59(C8)，28.84(C9)；C₁₇H₂₀NO⁺HRMS (ESI)⁺) The m/z calculation was 254.1537, and the value was found to be 254.1539.

6- (5-phenylpentyl) pyridine aldoxime 7:

acetaldehyde 6(150mg, 0.592mmol, 1 equiv.), hydroxylamine hydrochloride (82mg, 1.184mmol, 2 equiv.) and CH₃CO₂A solution of Na (146mg, 1.776mmol, 3 equiv.) in dry ethanol (12mL) was stirred at reflux for 16 h. After completion (monitored by TLC), solids were removed by filtration through a short pad of celiteThe material was removed, the solvent was evaporated, and the residue was purified by column chromatography (EtOAc/PE 1:9) to provide the desired oxime 7(135mg, 85%) as a white solid. R_f(20% EtOAc + PE) 0.40; IR (near Infrared) V_max 3080，2926，2856，1720，1575，1452，1269，986，780，699，658，569，458cm^-1；¹H NMR(400MHz,CDCl₃)δ(ppm)10.20(br s,1H,-OH),8.30(s,1H,H₁₈),7.59(br d,J＝8.0Hz,1H,H₃),7.50(t,J＝7.8Hz,1H,H₄),7.27-6.93(m,6H,H₅,H₁₃-H₁₇),2.74(t,J＝7.8Hz,2H,H₇),2.50(t,J＝7.5Hz,2H,H₁₁),1.68(quintet,J＝7.5,7.8Hz,2H,H₈),1.57(quintet,J＝7.5Hz,2H,H₁₀),1.32(quintet,J＝7.1,7.8Hz,2H,H₉)；¹³C NMR(100MHz,CDCl₃) δ (ppm)162.23, × 160.10(C6),151.36, × 150.73(C2),150.40(C18),142.61, × 142.23(C12), × 138.33,136.97(C4), × 129.46,128.33(C14, C16),128.16(C13, C17),125.61, × 125.53(C5), × 124.23,123.01(C15), × 120.84,118.112(C3),37.86, × 37.29(C7),35.75(C11),31.22, × 31.10(C10),29.82(C8),28.91, × 28.69(C9) (cis and trans mixture); c₁₇H₂₁N₂O⁺The calculated result of HRMS (ESI +) m/z of (D) was 269.1648, and the found value was 269.1670.

Synthesis of 6-pentadecylpyridinealdoxime 10：

6- (pentadecyl-1-yn-1-yl) pyridine aldoxime 9:

to THF/Et₃To a degassed solution of oxime 2(51mg, 0.252mmol, 1.05 equiv) in N (4mL/2mL) was added Pd [ PPh ]₃]₄(42mg, 0.036mmol, 0.15 equiv.) and CuI (14mg, 0.072mmol, 0.3 equiv.). After degassing the reaction mixture at room temperature for 5 minutes, alkyne 8 (5) was added dropwise0mg, 0.240mmol, 1 eq) and the reaction mixture was stirred at room temperature for 16 h. Upon completion (monitored by TLC), the reaction mixture was concentrated under reduced pressure and the residue was purified by column chromatography (EtOAc/PE6:94) to afford the desired coupled oxime 9(65mg, 83%) as a white solid. R_f(20% EtOAc + PE) 0.55; IR (near Infrared) Vmax 3179,3092,2914,2850,2226,1722,1567,1450,1268,1160,992,809,733,709,657,640,549,496cm^-1；¹H NMR(400MHz,CDCl₃)δ(ppm)8.50(s,1H,OH),8.26(s,1H,H₂₂),7.72(br d,J＝7.8Hz,1H,H₃),7.61(t,J＝7.8Hz,1H,H₄),7.23(br d,J＝7.7Hz,1H,H₅),2.41(t,J＝7.2Hz,2H,H₉),1.61(quintet,J＝7.2Hz,2H,H₁₀),1.41(m,2H,H₁₁),1.24(s,18H,H₁₂-H₂₀),0.85(t,J＝6.5Hz,3H,H₂₁)；¹³C NMR(100MHz,CDCl₃)δ(ppm)151.89(C2),150.73(C22),143.69(C6),136.64(C4),127.13(C5),125.92(C15),119.12(C3),92.10(C7),79.76(C8),31.91,29.64,29.49,29.35,29.13,29.00,28.31,22.68,19.40,14.11(C9-C21)；；C₂₁H₃₃N₂O₁ ⁺The calculated result of HRMS (ESI +) m/z of (D) was 329.2587, and the found value was 329.2549.

6-pentadecylpyridinealdoxime 10:

10% Pd/C (3mg, 0.027mmol, 0.25 equiv.) was added to a degassed solution of oxime 9(35mg, 0.107mmol, 1 equiv.) in dry EtOAc (2 mL). By H₂After three washes, at room temperature and at H₂(1atm.) the reaction mixture was stirred for 2 hours. Upon completion, the catalyst was removed by short column celite filtration, the solvent was evaporated, and the residue was purified by column chromatography (EtOAc/PE6:94) to provide oxime 3as a white solid (30mg, 85%); r_f(20% EtOAc + PE) 0.65; IR (near Infrared) v_max 3187,3083,2914,2849,1575,1457,1160,985,777,718,656,517,479cm^-1；¹H NMR(400MHz,CDCl₃)δ(ppm)8.36(br s,1H,OH),8.25(s,1H,H₂₂),7.61-6.54(m,2H,H₃,H₄),7.11(dd,J＝2.5,6.1Hz,1H,H₅),2.78(t,J＝7.2Hz,2H,H₇),1.70(m,2H,H₈),1.24(s,24H,H₁₂-H₂₀),0.86(t,J＝6.6Hz,1H,H₂₁)；¹³C NMR(100MHz,CDCl₃)δ(ppm)162.71(C6),151.12(C2),151.02(C23),136.73(C4),123.09(C5),118.21(C3),38.28,31.92,29.99,29.69,29.56,29.49,29.41,29.36,22.69,14.12(C7-C22)。C₂₁H₃₇N₂O⁺The calculated result of HRMS (ESI +) m/z of (D) was 333.2900, and the found value was 333.2918.

Synthesis of 2- ((hydroxyimino) methyl) -6- (pyridin-1-chloride-3-ylethynylene) pyridine-1-chloride 13:

3-fluoro-6- (pyridin-3-ynylidene) pyridine aldoxime 12:

to THF/Et₃Pd [ PPh ] was added to a degassed solution of oxime 2(211mg, 1.05mmol, 1.05 equiv.) in N (3mL/3mL)₃]₄(173mg, 0.15mmol, 0.15 equiv.) and CuI (57mg, 0.30mmol, 0.3 equiv.). After degassing the reaction mixture at room temperature for 5 minutes, the degassed alkyne 11(103mg, 1mmol, 1 eq) in dry THF (3mL) was added dropwise and the reaction mixture was stirred at room temperature for 16 hours. Upon completion (monitored by TLC), the reaction mixture was concentrated under reduced pressure and the residue was purified by column chromatography (EtOAc/PE 45:55) to afford the desired coupled oxime 12(200mg, 90%) as a white solid. R_f(60% EtOAc + PE) 0.35; IR (near Infrared) v_max3065,2764,1578,1561,1443,1288,1143,1042,990,981,798,697,637,563,499cm^-1；¹H NMR(400MHz,DMSO-d6)δ(ppm)11.84(s,1H,OH),8.83(br s,1H,H₁₄),8.65(br s,1H,H₁₂),8.11-8.04(m,2H,H₁₀,H₁₅),7.91(br t,J＝7.8Hz,1H,H₄),7.83(br d,J＝7.8Hz,1H,H₃),7.68(br d,J＝7.8Hz,1H,H₅),7.50(dd,J＝4.8,7.8Hz,1H,H₁₁)；¹³C NMR(100MHz,DMSO-d6)δ(ppm)152.855(C2),151.93(C14),149.70(C12),148.34(C15),141.51(C6),139.03(C10),137.65(C4),127.50(C5),123.78(C11),119.84(C3),118.48(C9),91.27(C7),85.48(C8)；C₁₃H₁₀N₃O⁺The calculated result of HRMS (ESI +) m/z of (D) was 224.0818, and the found value was 224.0840.

2- ((hydroxyimino) methyl) -6- (pyridin-1-chloride-3-ynylidene) pyridine-1-chloride 13:

to compound 12(40mg) in water (1mL) was added 1.2N HCl (1mL) and stirred for 2 minutes, and stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure to provide hydrochloride salt 13 as a white solid in quantitative yield. IR (near Infrared) v_max 3018,2970,2502,2080,1561,1465,1290,1005,813,726,674,548,499cm^-1；¹H NMR(400MHz,D₂O)δ(ppm)9.05(br s,1H),8.83(br d,J＝5.8Hz,1H),8.75(dt d,J＝1.6,8.3Hz,1H),8.19(s,1H),8.15-8.04(m,2H),7.87(dd,J＝0.6,8.0Hz,1H),7.81(br d,J＝0.6,7.8Hz,1H)；¹³C NMR(100MHz,D₂O)δ(ppm)150.85,149.74,148.30,144.75,142.04,141.28,139.9,130.03,128.12,123.99,122.81,92.14,85.20；C₁₃H₁₀N₃O⁺The calculated result of HRMS (ESI +) m/z of (D) was 224.0818, and the found value was 224.0826.

Synthesis of 6- (4- (naphthalen-1-ylamino) butyl) pyridine aldoxime 20:

2- (naphthylamino) -ethanol 15:

according to Couty et al¹Procedures for the synthesis of substituted aromatic amines; a solution of 1-iodonaphthalene 14(2.50g, 9.8mmol), 2-aminoethanol (1.78mL, 29.5mmol), copper chloride (132mg, 1.0mmol) and freshly pulverized KOH (1.10g, 19.7mmol) in dimethyl sulfoxide (2mL) was stirred at room temperature for 18 h. Adding saturated NH to the brownish red solution₄Aqueous Cl (5mL) and the solution was extracted (EtOAc, 3X 20 mL). Washed (brine, 20mL), dried (MgSO4), filtered and the combined extracts concentrated in vacuo. Silica gel chromatography (30% ethyl acetate in light petroleum ether) gave the title compound 15(1.68g, 9.0mmol) as a beige oil. IR (near Infrared) v_max 3404,3051,2973,2869,1581,785cm^-1；¹HNMR(400MHz,CDCl₃)δ(ppm)7.93-7.76(m,2H,ArH),7.52-7.29(m,4H,ArH),6.67(d,J＝7.3Hz,1H,CHCNH),4.01(t,J＝5.1Hz,2H,NHCH₂CH₂OH),3.49(t,J＝5.1Hz,2H,NHCH₂CH₂OH)；¹³C NMR(101MHz,CDCl₃)δ(ppm)142.8,134.6,128.7,126.4,125.9,125.0,123.9,120.0,118.5,105.5,61.0,46.6。

2- [ (2-hydroxyethyl) (naphthalen-1-yl) amino ] acetonitrile 16:

according to Couty et al¹Procedure for N-cyanomethylation of aromatic aminoethanol; a solution of 2- (naphthylamino) -ethanol 15(745mg, 4.0mmol) and paraformaldehyde (717mg, 8.0mmol) in MeCN (20mL) was heated to 90 ℃ for 18 hours. The white suspension was cooled to room temperature (rt) and TMSCN (1.06mL, 8.0mmol) and AcOH (0.46mL, 8.0mmol) were added to the reaction and the pale yellow reaction solution was stirred at 90 ℃ for 18 h. The reaction was cooled to room temperature and H was added₂O (40mL) and extract the water mixture (CH)₂Cl₂10 mL). The organic extract was washed with aqueous NaOH (1M, 20mL), brine (10mL) and dried (MgSO)₄) Filtered and concentrated in vacuoAn organic extract. Silica gel chromatography (30% ethyl acetate in light petroleum ether) afforded the title compound 16(850mg, 3.8mmol, 94% over two steps) as a colorless solid. mp 70-71 deg.C (literature value)¹71-73 ℃); IR (near Infrared) v_max 3422,3050,2956,2236,1705,1418,802cm^-1；¹HNMR(400MHz,CDCl₃)δ(ppm)8.17(d,J＝8.3Hz,1H,8-CH),7.91(d,J＝7.6Hz,1H,2-CH),7.75(d,J＝7.6Hz,1H,4-CH),7.61-7.42(m,4H,ArH),4.17(s,2H,CH₂CN),3.78(br t,J＝5.0,2H,NCH₂CH₂OH),3.54(t,J＝5.0Hz,2H,NCH₂CH₂OH)；¹³C NMR(101MHz,CDCl₃)δ(ppm)145.8,134.8,129.7,128.5,126.2,126.1,125.8,125.7,122.9,119.1,116.0,61.1,55.1,44.7。

1- (naphthalen-1-yl) azetidine-2-carbonitrile 17:

according to Couty et al¹A procedure for the production of aromatic azetidines; to 0 ℃ CH₂Cl₂2- [ (2-hydroxyethyl) (naphthalen-1-yl) amino group in (10mL)]Acetonitrile 16(500mg, 2.2mmol) and Et₃To a solution of N (0.77mL, 5.5mmol) was added MsCl (0.21mL, 2.6mmol) dropwise. The colorless reaction solution was stirred at 0 ℃ for 30 minutes and slowly warmed to room temperature. The reaction was stirred at room temperature for a further 30 minutes. Addition of H₂O (20mL), organic layer separated and aqueous layer extracted (CH)₂Cl₂20 mL). After drying (MgSO)₄) The combined extracts were washed with aqueous HCl (2M, 10mL) and brine (10mL) before being filtered and concentrated in vacuo. The pale yellow residue was taken directly to the next step and extracted in dry THF (15 mL). Adding to the solution at 0 deg.C^tBuOK (297mg, 2.6 mol). The reaction was allowed to slowly warm to room temperature and H was added₂O (20 mL). The solution was extracted (EtOAc, 3X 20mL), the combined organics were washed with brine (20mL), dried (MgSO)₄) Filtered and concentrated in vacuo. Silica gel chromatography (10% ethyl acetate in light petroleum ether) afforded the title compound as a colorless solidTitle compound 17(850mg, 3.8mmol, 94% yield over two steps). mp 129-¹130 ℃.); IR (near Infrared) v_max 3433,3045,2958,2248,1577,788cm^-1；¹HNMR(400MHz,CDCl₃)δ(ppm)7.96-7.82(m,2H,ArH),7.56-7.38(m,4H,ArH),6.75(d,J＝7.3Hz,1H,2-CH),4.93(dd,J＝8.3,6.6Hz,1H,NCH₂CH₂CHCN),4.51(ddd,J＝8.3,6.6,4.9Hz,1H,NCHHCH₂CHCN),3.88(dt,J＝8.3,6.8Hz,1H,NCHHCH₂CHCN),2.91-2.80(m,1H,2H,NCH₂CHHCHCN),2.78-2.66(m,1H,NCH₂CHHCHCN)；¹³C NMR(101MHz,CDCl₃)δ(ppm)145.0,134.7,128.6,126.1,125.6,125.2,125.1,122.9,122.5,118.4,109.5,54.3,51.0,22.7。

N- (3-butyl-1-yl) naphthylamine 18:

according to Couty et al²Procedures for the formation of aromatic homopropargylamines from aromatic azetidines; to a solution of 1- (naphthalen-1-yl) azetidine-2-carbonitrile 17(1.00g, 4.8mmol) in toluene (15mL) was added dibutyltin oxide (298mg, 1.2mmol) and TMSN₃(0.95mL, 7.2mmol) and the reaction stirred at 60 ℃ for 96 h. The brown reaction solution was cooled to room temperature and concentrated in vacuo. Silica gel chromatography (2% ethyl acetate in hexanes) gave title compound 18(454mg, 2.3mmol, 48%) as a colorless oil. IR (near Infrared) v_max 3293,3050,2975,2117,1690,767cm^-1；¹HNMR(400MHz,CDCl₃)δ(ppm)7.90-7.79(m,2H,5-CH,8-CH),7.57-7.29(m,4H,3-CH,4-CH,6-CH,7-CH),6.66(d,J＝7.3Hz,1H,2-CH),3.50(t,J＝6.5Hz,2H,NHCH₂CH₂CCH),2.69(td,J＝6.5,2.7Hz,2H,NHCH₂CH₂CCH),2.11(t,J＝2.7,1H,NHCH₂CH₂CCH)；¹³C NMR(101MHz,CDCl₃)δ(ppm)146.7,134.4,128.7,126.4,125.9,125.0,123.8,121.0,119.9,118.3,85.2,70.4,27.4,18.9。

N- (4- {6- [ (hydroxyimino) methyl ] pyridin-2-yl } but-3-yn-1-yl) naphthalen-1-amine 19:

to a solution of N- (3-but-1-yl) naphthylamine 18(400mg, 2.0mmol) in anhydrous THF/Et₃Pd (PPh) was added to a degassed solution in N (7mL/3mL)₃)₄(238mg, 0.2mmol) and CuI (78mg, 0.4 mmol). To the resulting orange reaction mixture was added dropwise a degassed solution of 6-bromopyridine aldoxime 2(453mg, 2.2mmol) in anhydrous THF (20 mL). The brown solution was stirred at room temperature for 16 hours. The reaction was concentrated in vacuo. Silica gel chromatography (hexane to 10% ethyl acetate in hexane) afforded the title compound 19(350mg, 54%) as an orange solid: mp is 143-144 ℃;¹HNMR(400MHz,CDCl₃)δ(ppm)8.28(s,1H,NOH),7.95-7.73(m,4H,ArH),7.67(t,J＝7.8Hz,1H,3-CH),7.53-7.33(m,4H,ArH),6.70(d,J＝7.8Hz,1H,2-CH),3.66(t,J＝6.7Hz,2H,NHCH₂CH₂),2.96(t,J＝6.7Hz,2H,NHCH₂CH₂)；¹³C NMR(101MHz,CDCl₃)δ(ppm)152.0,150.5,143.1,142.1,136.8,134.4,128.7,127.8,127.3,126.4,125.9,125.0,123.8,120.0,119.8,105.2,88.6,81.4,42.6,19.8；IR(neat)ν3350,3152,3047,2864,2645,2200cm^-1，C₂₀H₁₈N₃O⁺HRMS (ESI)⁺The calculated m/z was 316.1444, which was found to be 316.1445.

N- (4- {6- [ (hydroxyimino) methyl ] pyridin-2-yl } but-3-yn-1-yl) naphthalen-1-amine 20:

to N- (4- {6- [ (1E) - (hydroxyimino) methyl]To a degassed suspension of pyridin-2-yl } but-3-yn-1-yl) naphthalen-1-amine 19(173mg, 0.5mmol) in dry methanol (10mL) was added Pearlman's catalyst (77mg, 0.5 mmol). The reaction vessel was evacuated and flushed with hydrogen five times. The black reaction mixture was stirred at room temperature for 18 hours. The catalyst was removed by filtration through celite and under vacuumRemoving the solvent. Chromatography on silica gel (50% ethyl acetate in hexanes) afforded the title compound 20(60mg, 34%) as a colorless solid: mp is 151-; IR (near Infrared) v_max 3351,3047,2867,2642,lcm^-1；¹HNMR(400MHz,CDCl₃)δ(ppm)8.21(s,1H,NOH),7.77-7.67(m,2H,ArH),7.60-7.49(m,2H,ArH),7.41-7.30(m,2H,ArH),7.26(t,J＝8.2Hz,1H,NCCHCHCH),7.15(d,J＝8.2Hz,1H,NCCHCHCH),7.09(dd,J＝7.1,1.7Hz,1H,7-CH),6.53(d,J＝7.5Hz,1H,2-CH),3.26(t,J＝6.9Hz,2H,NHCH₂CH₂CH₂CH₂),2.85(t,J＝6.7Hz,2H,NHCH₂CH₂CH₂CH₂),2.00-1.65(m,4H,NHCH₂CH₂CH₂CH₂)；¹³C NMR(101MHz,CDCl₃)δ(ppm)162.0,151.1,151.0,143.4,137.1,134.3,128.6,126.6,125.7,124.7,123.4,123.3,119.9,118.6,117.3,104.4,44.1,37.7,28.8,27.6；C₂₀H₂₂N₃O⁺HRMS (ESI) + m/z calculated 320.1757, found 320.1759.

Reference material:

county et al, J.org.chem, 2016, 81, pages 2899-2910;

coutty et al, chem.Comms, 2016, 52, p.10072, 10075.

Synthesis of methyl 3-hydroxy-6- (4- (quinolin-4-ylamino) butyl) picolinate 26:

4-bromoquinoline 22:

according to Margolis et al¹Procedure for the synthesis of bromoquinoline; PBr is prepared from₃(3.34 mL, 35.5 mmol) was added dropwise to a solution of 4-quinolinol 21(5.00 g, 34.4 mmol) in DMF (50mL) at 60 ℃. After addition, the color was observed to change from yellowTurning bright orange with air bubbles. The orange reaction mixture was stirred at 45 ℃ for 45 minutes. The solution was cooled to room temperature and washed with H₂Diluted with O (20mL) and saturated NaHCO was added slowly₃Aqueous solution, basifying the reaction mixture to pH 10. By CH₂Cl₂(5X 20mL) of the extract solution, then combine the organic solutions and dilute with H₂O (20mL) and dried (MgSO)₄) Filtered and concentrated in vacuo. Silica gel chromatography (EtOAc) afforded title compound 22(5.56 g, 26.7 mmol, 78%) as a cream solid. The physical and spectral data were consistent with the reported values. mp-28-29 deg.C (literature value)²29.5-30.5 ℃); IR (near Infrared) v 3062,1615,1058 cm^–1；¹HNMR(400 MHz,CDCl₃)δ8.68(d,J＝4.6 Hz,1H,NCH),8.20(dd,J＝8.4,0.9Hz,1H,NCCHCHCHCH),8.11(d,J＝8.4 Hz,1H,NCCHCHCHCH),7.78(ddd,J＝8.4,7.0,1.4 Hz,1H,NCCHCH),7.71(d,J＝4.6 Hz,1H,NCHCH),7.66(ddd,J＝8.4,7.0,1.4Hz,1H,NCCHCHCHCH)；¹³C NMR(100MHz,CDCl₃)δ149.9,149.0,134.2,130.4,129.9,127.9,127.9,126.8,125.1。

N- (but-3-yn-1-yl) quinolin-4-amine 24:

according to Musonda et al³Procedure for the synthesis of alkylated quinolines; commercial 3-n-butyl-1-amine 23(4.72mL, 57.7mmol) was added to 4-bromoquinoline 22(3.00g, 14.4mmol) to form a thin cream-colored paste. The paste was heated to 80 ℃ for 1 hour without stirring. The temperature was raised to 140 ℃ and the paste was heated with stirring for 18 hours. The viscous brown reaction mixture was cooled to room temperature and purified by silica gel chromatography (20% MeOH in EtOAc) to provide the title compound 24 as a cream solid (2.82g, 14.4mmol, 100%): mp is 165-166 ℃; IR (near Infrared) v_max3281,3169,3067,1573,1151cm^–1；¹H NMR(400MHz,DMSO-d₆)δ8.45(d,J＝5.9Hz,1H,NCH),8.30(dd,J＝8.3,1.2Hz,1H,NCCHCHCHCH),7.93(br.s,1H,NH),7.82(dd,J＝8.3,1.2Hz,1H,NCCHCHCHCH),7.71(ddd,J＝8.3,7.0,1.2Hz,1H,NCCHCHCHCH),7.51(ddd,J＝8.3,7.0,1.2Hz,1H,NCCHCHCHCH),6.63(d,J＝5.9Hz,1H,NCHCH),3.54(q,J＝7.0Hz,2H,NHCH₂CH₂CCH),2.91(t,J＝2.7Hz,1H,NHCH₂CH₂CCH),2.58(td,J＝6.8,2.7Hz,2H,NHCH₂CH₂CCH)；¹³C NMR(100MHz,DMSO-d₆)δ151.3,148.2,145.1,130.2,126.4,124.7,122.1,118.1,98.3,82.1,72.6,41.4,17.8；C₁₃H₁₃N₂ ⁺HRMS (ESI)⁺The calculated m/z was 197.1073, which was found to be 197.1072.

6- (4- (quinolin-4-ylamino) but-1-yn-1-yl) pyridine aldoxime 25:

to N- (but-3-yn-1-yl) quinolin-4-amine 24(1.00g, 5.1mmol) in dry THF/Et₃Pd (PPh) was added to a degassed solution in N (50mL/15mL)₃)₄(588mg, 0.5mmol) and CuI (194mg, 1.0 mmol). To the resulting orange reaction mixture was added dropwise a degassed solution (20mL) of 6-bromopyridine aldoxime 2(1.13g, 5.6mmol) in anhydrous THF. The brown solution was stirred at room temperature for 18 hours. The reaction was concentrated in vacuo. Silica gel chromatography (20% MeOH in EtOAc) afforded the title compound 25(1.55g, 4.9mmol, 96%) as an orange solid: mp 202-; IR (near Infrared) v_max 3291,3068,2947,2241,1617,1222,1051cm^–1；¹H NMR(400MHz,DMSO-d₆)δ11.90-11.69(m,1H,CHNOH),8.43(br d,J＝5.4Hz,1H,NCH),8.23(d,J＝8.3Hz,1H,NCCH),8.03(s,1H,CHNOH),7.85-7.65(m,4H,ArH),7.52-7.38(m,3H,ArH),6.60(d,J＝5.4Hz,1H,NCHCH),3.61(q,J＝6.7Hz,2H,NHCH₂CH₂),2.88(t,J＝6.7Hz,2H,NHCH₂CH₂)；¹³C NMR(100MHz,DMSO-d₆)δ152.5,150.3,149.8,148.4,147.8,142.4,137.8,129.0,128.6,126.8,124.1,121.7,118.9,118.7,98.5,88.7,81.1,48.6,18.6；C₁₉H₁₇N₄O⁺HRMS (ESI)⁺Calculated m/z is 317.1397, foundThe value is 317.1396.

3-hydroxy-6- (4- (quinolin-4-ylamino) butyl) picolinic acid methyl ester 26:

to a degassed suspension of (E) -6- (4- (quinolin-4-ylamino) but-1-yn-1-yl) pyridylaldoxime 25(575mg, 1.8mmol) in dry methanol (20mL) was added a Pearlman catalyst (255mg, 1.8 mmol). The reaction vessel was evacuated and flushed with hydrogen five times. The black reaction mixture was stirred at room temperature for 18 hours. The catalyst was removed by filtration through celite, and the solvent was removed in vacuo to provide the title compound 26 as a milky solid (550mg, 1.7mmol, 94%): mp 218-219 deg.C; IR (near Infrared) v_max 3145,3026,2985,1593,1224,1026,658cm^–1；¹H NMR(400MHz,D₂O)δ8.15(s,1H,CHNOH),8.01(d,J＝7.1Hz,1H,NCH),7.89-7.49(m,7H,ArH),6.62(d,J＝7.1Hz,1H,NCHCH),3.54(t,J＝6.6Hz,2H,NHCH₂CH₂CH₂CH₂),3.09(t,J＝7.2Hz,2H,NHCH₂CH₂CH₂CH₂),1.99-1.76(m,4H,NHCH₂CH₂CH₂CH₂)；¹³C NMR(100MHz,D₂O)δ156.1,146.7,145.0,142.7,141.7,137.6,134.1,127.5,127.4,123.2,122.3,122.1,120.2,116.8,98.2,42.9,33.1,26.1,17.2；C₁₉H₂₁N₄O⁺HRMS (ESI)⁺The m/z calculation was 321.1710, and the value was found to be 3321.1713.

Reference material:

margolis, B.J. et al, J.org.chem, 2007, 72, p.2232-2235;

charette, A.B. et al, J.org.chem, 2017, 82, p. 5046-5067.

Musonda, C.C et al, bioorg.Med.chem.Lett., 2007, 17 th, p.4733-4736.

Synthesis of 6- (4- ((5-fluoroquinolin-4-yl) amino) butyl) pyridine aldoxime 31:

4-bromo-5-fluoroquinoline 28:

according to Kilpin-Guy et al¹Synthesis of substituted quinolines from anilines and Pulley et al²A modified procedure for the synthesis of bromoquinoline from quinalditol; meldrum's acid (9.49g, 65.9mmol) and triethyl orthoformate (21.3mL, 128.0mmol) were added to a solution of m-fluoroaniline 27(6.00g, 54.0mmol) in ethanol (100mL) at room temperature. The yellow solution was stirred at 90 ℃ for 2.5 hours. The solution was cooled to 0 ℃, and the resulting yellow solid was filtered and washed with cold ethanol (20 mL). The resulting pale yellow solid was dried and slowly added over 5 minutes to diphenyl ether (50mL) at reflux at 280 ℃. After addition, a large amount of white gas was observed and the colorless solution turned orange/brown. Reflux was maintained for 5 minutes and the reaction mixture was cooled to room temperature. During this time, the solution turned dark brown. To the solution was added petroleum ether (50mL) and the resulting brown crystals were isolated by filtration. Silica gel chromatography (10% methanol in EtOAc) afforded an inseparable mixture of 5-fluoroquinolin-4-ol and 7-fluoroquinolin-4-ol (about 9:1, by 19FNMR detection, 7.76g, 47.5 mmol). The mixture was directly taken to the next step.

To an inseparable mixture of 5-fluoroquinolin-4-ol and 7-fluoroquinolin-4-ol (4.00g, 24.5mmol) in DMF (30mL) was added phosphorus tribromide (1.86mL, 19.7mmol) at 60 ℃ and the mixture was stirred at 45 ℃ for 45 minutes. After cooling to room temperature, H was added₂O (25mL) and saturated Na was added₂CO₃Aqueous solution to adjust the pH of the solution to 10. Obtained H for crystals₂O (10mL) wash. Silica gel chromatography (25% acetoacetate in hexanes) gave only 4-bromo-5-fluoroquinoline 28(2.50g, 11.1mmol, 61% yield in three steps) as an orange solid. mp is 89-90 ℃; IR (near Infrared) v 3091,3041,1621cm^–1；¹HNMR(400MHz,CDCl₃)δ8.67(d,J＝4.6Hz,1H,NCH),8.22(dd,J＝9.2,5.9Hz,1H,NCCH),7.75(dd,J＝9.2,2.7Hz,1H,NCCHCHCH),7.68(d,J＝4.6Hz,1H,NCHCH),7.44(ddd,J＝9.2,5.9,2.7Hz,1H,NCCHCH)；¹³C NMR(100MHz,CDCl₃)δ161.9,150.6,149.6,133.7,128.9,124.7,124.1,118.0,113.1；¹⁹FNMR(376MHz,CDCl₃)δ108.7；C₉H₆BrFN⁺HRMS (ESI)⁺The calculated m/z was 225.9662, which was found to be 225.9658.

N- (but-3-yn-1-yl) -5-fluoroquinolin-4-amine 29:

according to the modified procedure of Musonda et al for the synthesis of alkylated quinolines³(ii) a 3-butyl-1-amine 23(0.55mL, 6.6mmol) was added to 4-bromo-5-fluoroquinoline 28(1.50g, 6.6mmol) to form an orange thin slurry. The slurry was heated to 100 ℃ and stirred for 18 hours. The reaction was further heated to 120 ℃ for 2 hours. The viscous, brown reaction mixture was cooled to room temperature and purified by silica gel chromatography (100% EtOAc to 20% MeOH in EtOAc) to provide the title compound 29 as a milky solid (1.06g, 4.9mmol, 75%). mp-225-226 ℃; IR (near Infrared) v_max 3079,2911,2240,1966cm^-1；¹HNMR(400MHz,DMSO-d₆)δ8.39(d,J＝5.4Hz,1H,NCH),8.27(dd,J＝10.8,8.3Hz,1H,NCCH),7.47(dd,J＝10.8,2.5Hz,1H,NCCHCHCH),7.44-7.39(m,1H,NHCH₂CH₂CCH),7.35(ddd,J＝10.8,8.3,2.5Hz,1H,NCCHCHCH),6.49(d,J＝5.4Hz,1H,NCHCH),3.45(q,J＝7.1Hz,2H,NHCH₂CH₂CCH),2.88(t,J＝2.7Hz,1H,NHCH₂CH₂CCH),2.55(td,J＝7.1,2.7Hz,2H,NHCH₂CH₂CCH)；¹³C NMR(100MHz,DMSO-d₆)δ163.4,160.9,151.9,149.7,124.5,115.8,113.5,112.2,98.2,82.2,72.4,41.2,17.7；¹⁹FNMR(376MHz,DMSO-d₆)δ112.1；C₁₃H₁₂FN₂ ⁺HRMS (ESI)⁺The calculated m/z was 215.0979, found to be 215.0983 Da.

6- (4- ((5-fluoroquinolin-4-yl) amino) but-1-yn-1-yl) pyridine aldoxime 30:

to N- (but-3-yn-1-yl) -5-fluoroquinolin-4-amine 29(0.50g, 2.3mmol) in dry THF/Et₃Pd (PPh) was added to a degassed solution in N (7mL/3mL)₃)₄(270mg, 0.2mmol) and CuI (89mg, 0.4 mmol). To the orange reaction mixture obtained was added dropwise a degassed solution of 6-bromopyridine aldoxime 2(516mg, 2.6mmol) in anhydrous THF (20 mL). The brown solution was stirred at room temperature for 16 hours. The reaction was concentrated in vacuo. Silica gel chromatography (EtOAc) showed title compound 30(140mg, 0.4mmol, 18%) as a creamy solid: 169 ℃ under mp & 168-; IR (near Infrared) v_max 3500,3435,3034,2960,2239,1966,1599,991cm^-1；¹H NMR(400MHz,DMSO-d₆)δ11.79(s,1H,CHNOH),8.42(d,J＝5.4Hz,1H,NCHCH),8.30(dd,J＝9.2,7.1Hz,1H,NCCHCHCHCF),8.02(s,1H,CHNOH),7.83-7.69(m,2H,ArH),7.53(t,J＝5.6Hz,1H,NHCH₂CH₂),7.48(dd,J＝10.8,2.7Hz,1H,NCCHCHCH),7.41(d,J＝7.1Hz,1H,NCCHCHCHCF),7.35(td,J＝8.7,2.7Hz,1H,NCCHCHCH),6.58(d,J＝5.4Hz,1H,NCHCH),3.66-3.51(m,2H,NHCH₂CH₂),2.86(t,J＝7.0Hz,2H,NHCH₂CH₂)；¹³C NMR(100MHz,DMSO-d₆)δ163.4,160.9,152.4,151.9,149.8,148.4,142.4,137.3,126.9,124.5,119.0,115.9,113.5,112.2,98.4,88.7,81.1,41.0,18.6；¹⁹FNMR(376MHz,DMSO-d₆)δ111.9；C₁₉H₁₆FN₄O⁺HRMS (ESI)⁺The calculated m/z was 335.1303, which was found to be 335.1298.

6- (4- ((5-fluoroquinolin-4-yl) amino) butyl) pyridine aldoxime 31:

to a degassed suspension of 6- (4- ((5-fluoroquinolin-4-yl) amino) but-1-yn-1-yl) pyridylaldoxime 30(50mg, 0.1mmol) in dry methanol (5mL) was added Pearlman's catalyst (21mg, 0.1 mmol). The reaction vessel was evacuated and flushed with hydrogen five times. The black reaction mixture was stirred at room temperature for 18 hours. The catalyst was removed by filtration through celite, and the solvent was removed in vacuo to provide the title compound 31(50mg, 0.1mmol, 99%) as a milky solid: mp 206-; IR (near Infrared) v_max 3247,2935,2859,1978,1584,806cm^–1；¹H NMR(400MHz,D₂O)δ8.44(t,J＝8.1Hz,1H,NCCHCHCH),8.19(d,J＝7.3Hz,1H,NCHCH),8.16-8.13(m,2H,CHNOH,NHCH₂CH₂CH₂CH₂),7.91-7.81(m,2H,NCCHCHCHCF,NCCHCHCHCF),7.70(d,J＝8.1Hz,1H,NCCHCHCH),7.42-7.32(m,2H,NCCHCHCH),6.70(d,J＝7.3Hz,NCHCH),3.58(t,J＝6.8Hz,2H,NHCH₂CH₂CH₂CH₂),3.12(t,J＝7.7Hz,2H,NHCH₂CH₂CH₂CH₂),2.01-1.76(m,4H,NHCH₂CH₂CH₂CH₂)；¹³C NMR(100MHz,D₂O)δ166.0,136.5,161.5,159.1,155.8,146.5,141.9,139.0,127.0,125.6,124.0,116.2,113.5,105.1,98.0,42.8,39.9,26.9,25.9；¹⁹FNMR(376MHz,D₂O)δ103.1；C₁₉H₂₂FN₄O⁺Of (ESI)⁺The calculated m/z was 339.1980, which was found to be 339.1980.

Reference material:

kiplin Guy R et al, bioorg.Med.chem.Lett., 2005, 15 th, p.1015-1018;

pulley et al, J.org.chem., 2007, 72, p.2232-2235;

musonda, c.c. et al, bioorg.med.chem.lett, 2007, 17 th, p.4733-4736.

Synthesis of 6- (4- ((8-methoxyquinolin-4-yl) amino) butyl) pyridine aldoxime 37:

8-methoxyquinolin-4-ol 33:

reprogramming according to Kilpin-Guy et al for the synthesis of substituted quinolines from anilines¹(ii) a Meldrum's acid (3.57g, 24.8mmol) and triethyl orthoformate (8.00mL, 48.0mmol) were added to a solution of o-anisidine 32(2.50g, 20.3mmol) in ethanol (20mL) at room temperature. The yellow solution was stirred at 90 ℃ for 2.5 hours. The solution was cooled to 0 ℃, and the resulting yellow solid was filtered and washed with cold ethanol (20 mL). The resulting pale yellow solid was dried and slowly added over 5 minutes to diphenyl ether (50mL) at reflux at 280 ℃. After addition, a large amount of white gas was observed and the colorless solution turned orange/brown. Reflux was maintained for 5 minutes and the reaction mixture was cooled to room temperature. During this time, the solution was dark brown. To the solution was added petroleum ether (50mL) and the resulting yellow crystals were isolated by filtration. Silica gel chromatography (5% methanol in EtOAc) afforded 8-methoxyquinolin-4-ol 33(7.76g, 47.5mmol) as a light orange solid: mp 168 deg.C (literature value)²168-; IR (near Infrared) v 2921,2851,1272,1041cm^-1；¹HNMR(400MHz,DMSO-d₆)δ(ppm)11.35(s,1H,OH),7.58-7.70(m,1H,NCH),7.38-7.43(m,1H,ArH),7.23-7.27(m,2H,OHCCCH,OMeCCH),6.95-7.08(m,1H,OHCCH),3.99(s,3H,OMe)；¹³C NMR(101MHz,DMSO-d₆)δ(ppm)177.1,149.0,139.3,130.5,123.2,119.1,116.7,111.4,109.6,56.6；C₁₀H₁₀NO₂ ⁺HRMS (ESI)⁺The calculated value of m/z was 176.0706, and found to be 176.0707.

4-bromo-8-methoxyquinoline 34:

following the reprogramming of Pulley et al for the synthesis of bromoquinolines from quinazolinols³(ii) a To a solution of 8-methoxyquinolin-4-ol 33(2.50g, 14.3mmol) in DMF (20mL) was added phosphorus tribromide (1.54mL, 16.4mmol) and the mixture was stirred at 45 ℃ for 45 minutes. After cooling to room temperature, H was added₂O (25mL) and saturated Na was added₂CO₃Aqueous solution to adjust the pH of the solution to 10. The resulting milky white crystals were filtered and combined with H₂O (10mL) gave 4-bromo-5-fluoroquinoline 34(2.58g, 10.8mmol, 76%) as a milky white solid. mp is 99-101 ℃; IR (near Infrared) v_max 1252,1085cm^-1；¹HNMR(400MHz,CDCl₃)δ(ppm)8.69(d,J＝4.6Hz,1H,NCH),7.78(d,J＝8.1Hz,1H,BrCCCH),7.75(d,J＝4.8Hz,1H,BrCCH),7.58(t,J＝8.2Hz,1H,ArH),7.14(d,J＝8.0Hz,1H,ArH),4.12(s,3H,OMe)；¹³C NMR(101MHz,CDCl₃)δ(ppm)155.5,148.5,134.1,129.0,128.0,125.8,118.5,108.5,56.3；C₁₀H₉BrNO⁺HRMS (ESI)⁺The calculated m/z was 237.9862, which was found to be 237.9865.

N- (but-3-yn-1-yl) -8-methoxyquinolin-4-amine 35:

according to the modified procedure of Musonda et al for the synthesis of alkylated quinolines⁴(ii) a 3-butyl-1-amine 23(1.20mL, 14.7mmol) was added to 4-bromo-8-methoxyquinoline 34(0.70g, 2.9mmol) to form an orange/yellow slurry. The slurry was heated to 80 ℃ for 1 hour without stirring. The temperature was raised to 100 ℃ by stirring for 18 hours. The viscous brown reaction mixture was cooled to room temperature and purified by alumina (basic) gel chromatography (10% MeOH in EtOAc) to give the title compound 35(0.80g, 2.9mmol, 99%) as a light orange solid. mp is 154-155 ℃; IR (near Infrared) v_max 3279,2938,2240,753cm^-1；¹HNMR(400MHz,DMSO-d₆)δ(ppm)8.36(d,J＝5.4Hz,1H,NCH),7.73(d,J＝7.8Hz,1H,MeOCCHCHCH),7.35(t,J＝8.1Hz,1H,MeOCCHCH),7.10(d,J＝7.3Hz,1H,OMeCCH),6.55(d,J＝5.4Hz,1H,NCHCH),3.91(s,3H,OMe),3.39-3.53(m,2H,CH₂CH₂CCH),2.89(t,J＝2.6Hz,1H,CH₂CH₂CCH),2.56(td,J＝7.1,2.7Hz,2H,CH₂CH₂CCH)；¹³CNMR(101MHz,DMSO-d₆)δ(ppm)155.2,149.9,148.8,139.6,124.3,119.6,113.3,108.5,99.1,82.5,72.6,55.8,41.5,18.0；C₁₄H₁₅N₂O⁺HRMS (ESI)⁺The calculated m/z was 227.1179, which was found to be 227.1183.

6- (4- ((8-methoxyquinolin-4-yl) amino) but-1-yn-1-yl) pyridine aldoxime 36:

to N- (but-3-yn-1-yl) -8-methoxyquinolin-4-amine 35(1.00g, 4.4mmol) in dry THF/Et₃Pd (PPh) was added to a degassed solution in N (7mL/3mL)₃)₄(511mg, 0.4mmol) and CuI (168mg, 0.9 mmol). To the orange reaction mixture obtained was added dropwise a degassed solution of 6-bromopyridine aldoxime 2(977mg, 4.8mmol) in anhydrous THF (20 mL). The brown solution was stirred at room temperature for 16 hours. The reaction was concentrated in vacuo. Chromatography on alumina (basic) gel (10% methanol in EtOAc) afforded the title compound 36(400mg, 1.1mmol, 26%) as a yellow solid: mp is 171-172 ℃; IR (near Infrared) v_max 3084,2900,2236,1617,1277,745cm^-1；¹H NMR(400MHz,DMSO-d₆)δ11.79(s,1H,CHNOH),8.35(d,J＝6.1Hz,1H,NCHCH),7.98(m,2H,NCC(OMe)CHCHCH,CHNOH),7.78(t,J＝7.8Hz,1H,NCCHCHCH),7.72(d,J＝7.8Hz,1H,NCCHCHCH),7.49(d,J＝8.2Hz,1H,NCC(OMe)CHCHCH),7.39(d,J＝7.8Hz,1H,NCCHCHCH),7.29(d,J＝8.2Hz,1H,NCC(OMe)CHCHCH),6.84(d,J＝6.1Hz,1H,NCHCH),3.99(s,3H,OMe),3.71(br t,J＝7.0,2H,NHCH₂CH₂),2.90(t,J＝7.0Hz,2H,NHCH₂CH₂)；¹³C NMR(100MHz,DMSO-d₆)δ152.6,152.5,152.1,148.3,145.1,142.3,137.3,134.2,126.8,125.4,119.0,118.5,113.7,110.3,99.0,88.3,81.3,56.1,48.5,18.7；C₂₀H₁₉N₄O₂ ⁺HRMS (ESI))⁺The calculated m/z was 347.1503, which was found to be 347.1506.

6- (4- ((8-methoxyquinolin-4-yl) amino) butyl) pyridine aldoxime 37:

to a degassed suspension of 6- (4- ((8-methoxyquinolin-4-yl) amino) but-1-yn-1-yl) pyridylaldoxime 36(110mg, 0.3mmol) in dry methanol (10mL) was added Pearlman's catalyst (9mg, 0.1 mmol). The reaction vessel was evacuated and flushed with hydrogen five times. The black reaction mixture was stirred at room temperature for 18 hours. The catalyst was removed by filtration through celite, and the solvent was removed in vacuo to provide the title compound 37(40mg, 0.1mmol, 36%) as a milky solid: mp is 107-108 ℃; IR (near Infrared) v_max 2927,1617,1581,980cm^–1；¹H NMR(400MHz,MeOD-d₆)δ8.29(d,J＝5.6Hz,1H,NCHCH),8.08(s,1H,CHNOH),7.71-7.57(m,3H,ArH),7.34(t,J＝8.2Hz,1H,NCC(OMe)CHCHCH),7.24-7.19(m,1H,NCC(OMe)CHCHCH),7.08(d,J＝7.8Hz,1H,NCCHCHCH),6.48(d,J＝5.6Hz,NCHCH),3.98(s,3H,OMe),3.36(t,J＝7.1Hz,2H,NHCH₂CH₂CH₂CH₂),2.83(t,J＝7.1Hz,2H,NHCH₂CH₂CH₂CH₂),1.94-1.66(m,4H,NHCH₂CH₂CH₂CH₂)；¹³C NMR(100MHz,MeOD-d₆)δ163.2,156.2,153.3,152.7,150.0,149.6,140.4,138.8,125.8,124.5,121.1,119.3,113.8,109.3,99.9,56.4,43.8,38.4,28.9,28.7；C₂₀H₂₃N₄O₂ ⁺HRMS (ESI)⁺The calculated m/z was 351.1816, which was found to be 351.1817.

Reference material:

kiplin Guy, R. et al, bioorg.Med.chem.Lett., 2005, 15 th page 1015-1018.

Lauer et al, j.am.chem.soc.,1946, 68 th, page 1268.

Pulley et al, J.org.chem.2007, 72 th, pp 2232-2235.

Musonda, c.c.; little, s.; yardley, v.; chibale, K., bioorg, Med, chem, Lett, 2007, No. 17, pages 4733-

Synthesis of 6- (3- (4-benzylpiperazin-1-yl) propyl) pyridine aldoxime 43:

n-benzylpiperazine 39:

the procedure for the synthesis of benzylpiperazine was followed by Bozell and Biannic¹(ii) a Piperazine 38(12.9g, 149.0mmol) in anhydrous CH at 0 deg.C₂Cl₂To the solution in (100mL) was added benzyl bromide (3.56mL, 29.8mmol) dropwise. The reaction was stirred at 0 ℃ for 1 hour. With saturated NaHCO₃The light yellow solution was washed with aqueous solution (2 solution (Omm)), dried (Na)₂SO₄) Filtered and concentrated in vacuo. Absolute ethanol was added and the white precipitate was filtered from the solution. The solution was concentrated in vacuo to afford title compound 39(24.8g, 141.0mmol, 94%) as a viscous yellow oil. IR (near Infrared) v_max 3289,2990,2960,2120,1120cm^-1；¹HNMR(400MHz,CDCl₃)δ(ppm)7.36-7.28(m,5H,ArH),3.50(s,2H,CH₂Ph),2.90(t,J＝4.9Hz,4H,CH₂N(Bn)CH₂),2.55-2.30(m,5H,CH₂NHCH₂)；¹³C NMR(101MHz,CDCl₃)δ(ppm)138.1,129.2,128.2,127.0,63.7,54.5,46.1。

1-benzyl-4-propynyl piperazine 41:

following the procedure of Corey, M for propargyl substitution of piperazine²(ii) a N-benzylpiperazine 39(1.75g, 9.93mmol), propargyl bromide 40 (80% in toluene) (1.28mL, 1)4.9mmol) and DIPEA (3.28mL, 19.9mmol) in CH₂Cl₂The solution in (50mL) was stirred at room temperature for 18 hours. Addition of H₂O (30mL) and the aqueous phase was separated and extracted (3 and separated and extracted). The combined organic layers were washed (brine, 30mL), dried (Na)₂SO₄) Filtered and concentrated in vacuo. Silica gel chromatography (50% ethyl acetate in hexanes) gave title compound 41(2.04g, 9.5mmol, 96%) as an orange oil. IR (near Infrared) v_max 3290,3026,2933,2807,2117,697cm^-1；¹HNMR(400MHz,CDCl₃)δ(ppm)7.37-7.28(m,5H,ArH),3.56(s,2H,PhCH₂),3.30(d,J＝2.5Hz,2H,NCH₂CCH),2.73-2.44(m,8H,PizCH₂),2.25(t,J＝2.5Hz,1H,NCH₂CCH)；¹³CNMR(101MHz,CDCl₃)δ(ppm)129.5,129.3,128.3,127.2,78.9,73.20,62.8,52.8,51.7,46.8；C_xH_xN_x ⁺HRMS (ESI)⁺The calculated m/z was 215.1543, which was found to be 215.1542.

6- (3- (4-benzylpiperazin-1-yl) prop-1-yn-1-yl) pyridine aldoxime 42:

to 1-benzyl-4-propynyl piperazine 41(1.30g, 6.1mmol) in anhydrous THF/Et₃Pd (PPh) was added to a degassed solution in N (7mL/3mL)₃)₄(0.70g, 0.6mmol) and CuI (0.23g, 1.2 mmol). To the orange reaction mixture obtained was added dropwise a degassed solution of 6-bromopyridine aldoxime 2(1.34g, 6.7mmol) in anhydrous THF (20 mL). The brown solution was stirred at room temperature for 18 hours. The reaction was concentrated in vacuo. Silica gel chromatography (EtOAc) showed title compound 42(750mg, 2.2mmol, 37%) as a cream solid: mp 143-; IR (near Infrared) v_max 3150,3048,2944,2808,2364,734cm^-1；¹HNMR(400MHz,CDCl₃)δ(ppm)12.17(s,1H,NOH),7.99(s,1H,CHNOH),7.71(dd,J＝8.0,1.0Hz,1H,NCCHCHCH),7.53(t,J＝8.0Hz,1H,NCCHCHCH),7.43-7.20(m,6H,ArH),3.70(s,2H,NCH₂CC),3.63(s,2H,PhCH₂),3.08-2.36(m,8H,PizCH₂)；¹³C NMR(101MHz,CDCl₃)δ(ppm)152.7,149.0,142.2,136.4,136.3,129.9,128.4,127.6,127.1,119.1,85.3,84.3,63.2,52.9,50.5,47.2；C₂₀H₂₃N₄O⁺HRMS (ESI)⁺The calculated value of m/z was 335.1866, and found to be 335.1863.

6- (3- (4-benzylpiperazin-1-yl) propyl) pyridine aldoxime 43:

to a degassed suspension of 6- (3- (4-benzylpiperazin-1-yl) propyl-1-yn-1-yl) pyridylaldoxime 42(200mg, 0.6mmol) in anhydrous methanol (10mL) was added Pearlman catalyst (44mg, 0.3 mmol). The reaction vessel was evacuated and flushed with hydrogen five times. The black reaction mixture was stirred at room temperature for 2 hours. The catalyst was removed by filtration through celite and the solvent was removed in vacuo. Silica gel Chromatography (CH)₂Cl₂To 10% methanol in CH₂Cl₂Middle) gave the title compound 43 (53%) as a pale yellow oil. IR near infrared) v_max 3162,3057,2939,2816,808cm^-1；¹HNMR(400MHz,CDCl₃)δ(ppm)8.19(s,1H,CHNOH),7.58-7.51(m,2H,ArH),7.36-7.27(m,5H,ArH),7.10(dd,J＝6.5,2.2Hz,1H,NCCHCHCH),3.55(s,2H,PhCH₂),2.82(t,J＝7.8Hz,1H,NCH₂CH₂CH₂),2.72-2.35(m,10H,PizCH₂,NCH₂CH₂CH₂),2.02(quin,J＝7.8Hz,NCH₂CH₂CH₂)；¹³C NMR(101MHz,CDCl₃)δ(ppm)161.3,151.8,150.3,137.6,136.6,129.4,128.3,127.2,122.8,118.0,62.9,57.7,25.8,25.5,35.8,26.3；C₂₀H₂₇N₄O⁺HRMS (ESI)⁺The calculated m/z was 339.2179, which was found to be 339.2176.

Reference to the literature

Bozell.J.J.et al, org.Lett.2013, 15 th, page 2730-2733.

Corey, M. et al, WO2017/184996A1 (2017).

Synthesis of 6- (3- (4-benzylpiperazin-1-yl) propyl) pyridine aldoxime 48:

3-butyl p-toluenesulfonate 45:

according to Winssinger et al¹Procedures for the formation of p-toluenesulfonyl protected alcohol; a solution of TsCl (8.98g, 47.1mmol) was added dropwise to CH at 0 deg.C₂Cl₂3-n-butan-1-ol 44(3.00g, 42.8mmol), DMAP (522mg, 4.3mmol) and Et (15mL)₃N (55.6mL, 7.70mL mmol). The yellow reaction solution was warmed to room temperature and stirred for 2 hours. Addition of H₂O (30mL), and the reaction was stirred at room temperature for 20 minutes. The organic layer was separated and the aqueous layer (CH) was extracted₂Cl₂5H2Cl2 and). Drying the combined extracts (Na)₂SO₄) Filtered and concentrated in vacuo. This afforded title compound 45(9.60g, 42.8mmol, 100%) as a red/brown oil. IR (near Infrared) v_max 3433,3045,2958,2248,1577,788cm^-1；¹HNMR(400MHz,CDCl₃)δ(ppm)7.81(d,J＝8.3Hz,2H,ArH),7.36(d,J＝8.3Hz,2H,ArH),4.11(t,J＝7.1Hz,2H,TsOCH₂CH₂CCH),2.56(td,J＝7.1,2.7Hz,2H,TsOCH₂TsOCH₂CH₂CCH),2.46(s,3H,PhCH₃),1.98(t,J＝2.7Hz,1H,TsOCH₂CH₂CCH)；¹³CNMR(101MHz,CDCl₃)δ(ppm)145.0,132.8,129.9,128.0,78.3,70.7,67.4,21.6,19.4。

1-benzyl-4- (butyl-3-yn-1-yl) piperazine 46:

according to Guarna et al²Procedures for forming alkylated piperazines; mixing Na₂CO₃(1.20g, 11.3mmol) and N-benzylpiperazine 39(2.00g, 11.3mmol) were added to a solution of 3-butyl-p-toluenesulfonate 45(2.30mL, 10.3mmol) in DMF (60 mL). The orange solution was stirred at 80 ℃ overnight. By H₂The reaction mixture was quenched O (10mL) and diethyl ether (10mL) was added. Separating the organic layer with H₂O (52O from organic layer), brine (10mL) and dried (Na)₂SO₄) Filtered and concentrated in vacuo. Silica gel chromatography (100% CH)₂Cl₂To 10% MeOH in CH₂Cl₂To yield the title compound 46(1.70g, 7.4mmol, 72%) as an orange oil. IR (near Infrared) v_max 3291,3026,2939,2807,2119,1676,697cm^-1；¹HNMR(400MHz,CDCl₃)δ(ppm)7.35-7.28(m,5H,ArH),3.52(s,2H,PhCH₂),2.61(t,J＝7.6Hz,2H,NCH₂CH₂CCH),2.58-2.42(m,8H,PizCH₂),2.41-2.34(m,2H,NCH₂CH₂CCH),1.97(t,J＝2.7Hz,1H,NCH₂CH₂CCH)；¹³CNMR(101MHz,CDCl₃)δ(ppm)138.1,129.2,128.2,127.0,82.8,69.0,63.0,57.0,52.9,52.8,16.8；C_xH_xN_x ⁺HRMS (ESI)⁺The calculated m/z was 229.1699, which was found to be 229.1695.

6- (4- (4-benzylpiperazin-1-yl) but-1-yn-1-yl) pyridylaldoxime 47:

to 1-benzyl-4- (butyl-3-yn-1-yl) piperazine 46(1.55g, 6.8mmol) in dry THF/Et₃Pd (PPh) was added to a degassed solution in N (7mL/3mL)₃)₄(1.16g, 0.7mmol) and CuI (0.19g, 1.4 mmol). To the orange reaction mixture obtained was added dropwise a degassed solution of 6-bromopyridine aldoxime 2(1.50g, 7.47mmol) in anhydrous THF (20 mL). The brown solution was stirred at room temperature for 18 hours. The reaction was concentrated in vacuo. Silica gel chromatography (50% acetoacetate in hexanes to acetoacetate) gave the title as a cream solidCompound 47(150mg, 0.4mmol, 6%). mp is 134-136 ℃; IR (near Infrared) v_max 3161,3060,2954,2808,2231,740cm^-1；¹HNMR(400MHz,CDCl₃)δ(ppm)11.48(s,1H,NOH),8.20(s,1H,CHNOH),7.76(dd,J＝8.0,1.0Hz,1H,NCCHCHCH),7.59(t,J＝8.0Hz,1H,NCCHCHCH),7.35-7.27(m,6H,ArH),3.56(s,2H,PhCH₂),2.82-2.41(m,12H,PizCH₂,NCH₂CH₂CC)；¹³C NMR(101MHz,CDCl₃)δ(ppm)152.9,149.7,143.1,137.4,136.5,129.5,128.3,127.3,126.7,119.1,89.1,80.8,63.0,56.6,52.6,52.5,17.4；C₂₁H₂₅N₄O⁺HRMS (ESI)⁺The calculated m/z was 349.2023, which was found to be 349.2018.

Reference to the literature

Winssinger et al, chem.comms., 2010, 46, page 5476-5478.

Guarna et al, J.Med.chem, 2010, 53 th, page 7119-7128.

6- (4- (4-benzylpiperazin-1-yl) butyl) pyridine aldoxime 48:

to a degassed suspension of 6- (4- (4-benzylpiperazin-1-yl) but-1-yn-1-yl) pyridylaldoxime 47(60mg, 0.2mmol) in anhydrous methanol (5mL) was added palladium (10% on carbon, 4mg, 0.04 mmol). The reaction vessel was evacuated and flushed with hydrogen five times. The black reaction mixture was stirred at room temperature for 1.5 hours. The catalyst was removed by filtration through celite and the solvent was removed in vacuo to afford title compound 48(53mg, 0.2mmol, 87%) as a yellow oil. IR (near Infrared) v_max 3181,3060,2938,2818,791cm^-1,¹HNMR(400MHz,CDCl₃)δ(ppm)8.18(s,1H,CHNOH),7.62-7.50(m,2H,ArH),7.34-7.29(m,5H,ArH),7.11(dd,J＝7.3,1.4Hz,1H,NCCHCHCH),5.31(s,2H,PhCH₂),2.82(t,J＝7.3Hz,2H,NCH₂CH₂CH₂CH₂),2.59-2.31(m,10H,PizCH₂,NCH₂CH₂CH₂CH₂),1.76(quin,J＝7.3Hz,NCH₂CH₂CH₂CH₂),1.58(br s,2H,NCH₂CH₂CH₂CH₂)；¹³C NMR(101MHz,CDCl₃)δ(ppm)161.9,151.5,150.6,136.7,136.6,129.3,128.2,127.1,122.9,118.1,63.0,58.4,53.0,52.7,37.9,27.6,26.1；C₂₀H₂₇N₄O⁺HRMS (ESI)⁺The calculated m/z was 353.2336, which was found to be 353.2332.

Synthesis of 6- (4- (3, 7-dimethyl-2, 6-dioxy-2, 3,6, 7-tetrahydro-1H-purin-1-yl) but-1-yn-1-yl) pyridine aldoxime 51:

1- (but-3-yn-1-yl) -3, 7-dimethyldihydro-1H-purine-2, 6-dione 50:

according toWait for¹An improved procedure for the synthesis of substituted amines by Mitsunobu reaction; to a solution of 3-butyl-1-ol 44(0.10ml, 1.4mmol), theobromine 49(500mg, 2.8mmol) and triphenylphosphine (728mg, 2.8mmol) in THF (15ml) was added ADDP (700mg, 2.8mmol) at room temperature. The yellow reaction was heated to 60 ℃ for 24 hours. Diluted cream color reaction solution (H)₂O, 50mL), aqueous solution was extracted (EtOAc, 3 ioac solution). The combined organic layers were dried (MgSO)₄) Filtered and concentrated in vacuo. The white residue was purified by silica gel chromatography (EtOAc) to give 1- (but-3-yn-1-yl) -3, 7-dimethyl-1H-purine-2, 6-dione 50(120mg, 0.5mmol, 37%) as a white solid: mp 192-; IR (near Infrared) v_max 3228,3107,2951,1697,1651cm^-1；¹HNMR(400MHz,DMSO-d₆)δ(ppm)8.00(s,1H,NCHN),3.99(t,J＝7.6Hz,2H,NCH₂CH₂CCH),3.87(s,3H,NCHN(CH₃)),3.40(s,3H,NCON(CH₃)),2.84(t,J＝2.7Hz,1H,NCH₂CH₂CCH),2.45(td,J＝7.6,2.7Hz,1H,NCH₂CH₂CCH)；¹³C NMR(101MHz,DMSO-d₆)δ(ppm)154.6,151.1,148.7,143.5,107.0,81.5,73.0,40.6,33.6,29.8,17.3；C₁₁H₁₃N₄O₂ ⁺HRMS (ESI)⁺Calculated m/z is 233.1033, found to be 233.1035, and C₁₁H₁₂N₄NaO₂ ⁺Calculated m/z of 255.0852, found a value of 255.0857.

6- (4- (3, 7-dimethyl-2, 6-dioxy-2, 3,6, 7-tetrahydro-1H-purin-1-yl) but-1-yn-1-yl) pyridine aldoxime 51:

to 1- (but-3-yn-1-yl) -3, 7-dimethyldihydro-1H-purine-2, 6-dione 50(200mg, 0.9mmol) in anhydrous THF/Et₃Pd (PPh) was added to a degassed solution in N (7mL/3mL)₃)₄(99mg, 0.1mmol) and CuI (33mg, 0.2 mmol). To the orange reaction mixture obtained was added dropwise a degassed solution of 6-bromopyridine aldoxime 2(190mg, 1.0mmol) in anhydrous THF (10 mL). The brown solution was stirred at room temperature for 16 hours. The reaction was concentrated in vacuo to give an orange solid as crude product. Silica gel chromatography (EtOAc to 10% MeOH in EtOAc) afforded the title compound 51(111mg, 0.3mmol, 36%) as colorless: mp is 210-211 ℃; IR (near Infrared) v_max 3178,3087,2872,2230,1700,1647,759cm^-1,¹H NMR(400MHz,DMSO-d₆)δ11.75(s,1H,CHNOH),8.03(s,1H,CHNOH),8.00(s,1H,NCHN),7.79(t,J＝8.1Hz,1H,NCCHCHCH),7.73(dd,J＝8.1,1.0Hz,1H,NCCHCHCH),7.39(dd,J＝8.1,1.0Hz,1H,NCCHCHCH),4.12(t,J＝7.5Hz,2H,NHCH₂CH₂),3.88(s,3H,NCHN(CH₃),3.42(s,3H,NCON(CH3),2.75(t,J＝7.5Hz,2H,NHCH₂CH₂)；¹³C NMR(100MHz,DMSO-d₆)δ154.2,152.4,150.7,148.4,148.3,143.1,142.3,137.3,126.8,119.0,118.9,106.5,87.4,81.3,33.2,29.4,17.7；C₁₇H₁₇N₆O₃ ⁺HRMS (ESI)⁺The calculated m/z was 353.1357, which was found to be 353.1358.

Reference to the literature

1.S. et al, Tet.letters, 1993, 34 th, 1639-1642.

Synthesis of II-bifunctional ethylcarbazole analogs

(3aS,4S,6R,6aR) -6- (6-amino-9H-purin-9-yl) -N- (but-3-yn-1-yl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolane-4-carboxamide 54:

(3aS,4S,6R,6aR) -6- (6-amino-9H-purin-9-yl) -2, 2-dimethyltetrahydrofuran [3,4-d ]][1,3]Synthesis of dioxolane-4-carboxylic acid 53 by using Debnath, J¹By the improved procedure of (1).

To a stirred solution of acid 53(500mg, 1.56mmol, 1 equiv.) in dry pyridine (15mL) was added 1-amino-3-butene 22(140mL, 1.71mmol, 1.1 equiv.) and EDCI (598mg, 3.12mmol, 2 equiv.) successively and the reaction mixture was stirred at room temperature under a nitrogen atmosphere overnight. Upon completion, the reaction mixture was directly concentrated under reduced pressure and the residue was purified by column chromatography (EtOAc to EtOAc/MeOH 95:5) to afford the desired amide 54 as a pale yellow solid (500mg, 80%). R_f(neat EtOAc) 0.18; IR (near Infrared) v_max 3289,3142,1672,1601,1526,1206,1090,1058,868,789,645,514cm^-1；¹H NMR(400MHz,CDCl₃)δ(ppm)8.32(s,1H),7.89(s,1H),7.28(m,1H),6.33(s,2H),6.13(d,J＝2.5Hz,1H),5.47(dd,J＝2.1,6.2Hz,1H),5.39(dd,J＝2.5,6.2Hz,1H),4.74(d,J＝2.1Hz,1H),3.21(m,2H),2.21(m,1H),2.10(m,1H),1.82(t,J＝2.6Hz,1H),1.63(s,3H),1.40(s,3H)；¹³C NMR(100MHz,CDCl₃)δ(ppm)168.98,155.86,153.11,148.99,139.82,120.21,114.37,91.70,86.33,83.59,82.88,80.81,69.83,37.49,26.95,25.07,18.85；C₁₇H₂₁N₆O₄ ⁺HRMS (ESI)⁺) The calculated m/z was 373.1575, which was found to be 373.1619.

(3aS,4S,6R,6aR) -6- (6-amino-9H-purin-9-yl) -N- (4- (6-formylpyridin-2-yl) but-3-yn-1-yl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolane-4-carboxamide 55:

pd [ PPh ] is added₃]₄(233mg, 0.202mmol, 0.15 equiv.) and CuI (77mg, 0.403mmol, 0.3 equiv.) are added to THF/Et₃Methyl 6-bromopyridylaldehyde 1(275mg, 1.478mmol, 1.1 equiv.) in N (10mL/8mL) was degassed. After degassing the reaction mixture at room temperature for 5 minutes, a degassed solution of alkyne 54(500mg, 1.344mmol, 1 eq) in THF (10mL) was added dropwise and the reaction mixture was stirred at room temperature for 16 hours. Upon completion, the reaction mixture was concentrated under reduced pressure and the residue was purified by column chromatography (pure EtOAc to EtOAc/MeOH 9:1) to afford the desired coupled pyridylaldehyde 55(510mg, 80%) as a concentrated syrup. IR (near Infrared) v_max 3318,1638,1582,1452,1209,1078,868,797,646,509cm^-1；¹H NMR(400MHz,CDCl₃)δ(ppm)9.90(s,1H),8.24(s,1H),7.85-7.65(m,3H),7.38(m,1H),6.40(s,1H),6.02(s,1H),5.33(s,2H),4.70(s,1H),3.35(m,2H),2.60-2.32(m,2H),1.57(s,3H),1.31(s,3H)；¹³C NMR(100MHz,CDCl₃)δ(ppm)192.88,169.13,155.67,153.12,152.63,148.97,143.63,139.87,137.20,130.85,120.19,114.62,91.94,88.87,85.74,83.55,82.54,80.66,37.41,27.09,25.11,20.08；C₂₃H₂₄N₇O₅ ⁺HRMS (ESI)⁺) The calculation result of m/z was 478.1806, and the value was found to be 478.1833.

(3aS,4S,6R,6aR) -6- (6-amino-9H-purin-9-yl) -N- (4- (6- (hydroxyimino) methyl) pyridin-2-yl) but-3-yn-1-yl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolane-4-carboxamide 56:

the following solutions were stirred at reflux for 16 hours: pyridine aldehyde 55(80mg, 0.168mmol, 1 equiv.), hydroxylamine hydrochloride (23mg, 0.336mmol, 2 equiv.), and CH₃CO₂A solution of Na (41mg, 0.503mmol, 3 equiv.) in dry ethanol (5 mL). After concentration under reduced pressure, with CH₂Cl₂The crude product was washed (5 x 10mL) to remove all impurities. The compound present in the round-bottom flask was pyridine aldoxime 56, dried under high vacuum (82mg, quantitative yield), and purified by¹H NMR confirmed. R_f(EtOAc); IR (near Infrared) v_max 3186,2925,1643,1579,1207,1089,980,867,797,726,649,509cm^-1；¹H NMR(400MHz,CDCl₃)δ(ppm)8.20(s,1H),8.11(s,1H),7.92(m,2H),7.46(m,2H),7.02(m,3H),6.05(d,J＝2.7Hz,1H),5.35(dd,J＝2.0,6.2Hz,1H),5.29(dd,J＝2.7,6.2Hz,1H),4.74(d,J＝2.0Hz,1H),3.41(m,2H),2.57-2.33(m,2H),1.59(s,3H),1.433(s,3H)；¹³C NMR(100MHz,CDCl₃)δ(ppm)169.14,155.54,152.89,152.05,149.16,148.60,142.61,139.82,136.66,126.72,119.58,119.47,114.74,91.86,88.15,85.78,83.45,82.68,81.13,37.45,27.11,25.14,20.21；C₂₃H₂₅N₈O₅ ⁺HRMS (ESI)⁺) The m/z calculation was 493.1901, and the value was found to be 493.1942.

(2S,3S,4R,5R) -5- (6-amino-9H-purin-9-yl) -3, 4-dihydroxy-N- (4- (6- (hydroxyimino) methyl) pyridin-2-yl) but-3-yn-1-yl) tetrahydrofuran-2-carboxamide 57:

1.2N HCl (185. mu.L, 0.610mmol, 1)0 equiv.) was added to a stirred solution of oxime 56(30mg, 0.061mmol, 1 equiv.) in dry methanol (5mL) and the reaction mixture was stirred at 55 ℃ for 5 hours. After completion, the reaction mixture was directly concentrated under reduced pressure and purified by reverse phase column chromatography (MeOH/H)₂O1:4) to provide salt 57 as a white solid in quantitative yield. IR (near Infrared) v_max3192,2927,1644,1580,1448,1305,1254,1045,989,808,726,642,533cm^-1；¹HNMR(400MHz,CD₃OD)δ(ppm)8.26(s,1H),8.25(s,1H),7.90(s,1H),7.68(br d,J＝7.8Hz,1H),7.0(t,J＝7.8Hz,1H),7.13(d,J＝7.5Hz,1H),6.30(d,J＝7.9Hz,1H),4.85(s,1H),4.54(s,1H),4.39(br d,J＝4.3Hz,1H),3.63(m,2H),2.74(m,2H)；¹³C NMR(100MHz,CD₃OD)δ(ppm)172.76,157.47,154.03,153.96,150.15,149.49,143.97,142.67,138.49,128.24,121.22,120.73,90.84,89.91,86.78,82.11,75.30,73.56,38.93,21.02；C₂₃H₂₅N₈O₅ ⁺HRMS (ESI)⁺) The m/z calculation was 493.1901, and the value was found to be 493.1942.

Reference to the literature

Debnath, j, et al, bioorg.med.chem., 2010, 18 th, page 8257-8263.

Synthesis of III-bifunctional pseudoethylcarbazole analogs

N- (9- ((3aR,4R, 6aR) -6- (((3- (6-formylpyridin-2-yl) prop-2-yn-1-yl) oxy) methyl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolan-4-yl) -9H-purin-6-yl) benzamide 59:

n- (9- ((3aR,4R,6R,6aR) -2, 2-dimethyl-6- ((prop-2-yn-1-yloxy) methyl) -tetrahydrofuran [3,4-d][1,3]Dioxolan-4-yl) -9H-purin-6-yl) benzamide 58 was synthesized by using Silvia, F¹According to known procedures.

To THF/Et₃Pd [ PPh ] was added to a degassed solution of 6-bromopyridylaldehyde 1(91mg, 0.490mmol, 1.1 equiv.) in N (3mL/2mL)₃]₄(77mg, 0.067mmol, 0.15 equiv.) and CuI (25mg, 0.134mmol, 0.3 equiv.). After degassing the reaction mixture at room temperature for 5 minutes, a degassed solution of alkyne 58(200mg, 0.445mmol, 1 eq) in THF (3mL) was added dropwise and the reaction mixture was stirred at room temperature for 16 hours. Upon completion, the reaction mixture was concentrated under reduced pressure and the residue was purified by column chromatography (EtOAc/petroleum ether 4:1) to afford the desired coupled pyridylaldehyde 59(200mg, 81%) as a concentrated syrup. IR (near Infrared) v_max 2935,1704,1609,1580,1452,1248,1210,1074,864,709,645,541cm^-1；¹H NMR(400MHz,CDCl₃)δ(ppm)9.97(s,1H),8.76(s,1H),8.32(s,1H),7.94(d,J＝7.6Hz,2H),7.85-7.77(m,2H),7.63-7.51(m,2H),7.45(t,J＝7.6Hz,2H),6.25(d,J＝2.1Hz,2H),5.32(dd,J＝2.1,6.2Hz,1H),5.03(dd,J＝2.1,6.2Hz,1H),4.56(q,J＝2.9Hz,1H),4.37(s,2H),3.89-3.77(m,2H),1.61(s,3H),1.37(s,3H)；¹³C NMR(100MHz,CDCl₃)δ(ppm)192.58,164.85,152.75,152.59,151.49,149.28,142.85,141.91,137.53,13335,132.81,131.23,128.78,127.81,123.24,120.91,114.32,91.61,85.94,85.47,85.20,84.55,70.32,59.09,27.12,25.28；C₂₉H₂₇N₆O₆ ⁺HRMS (ESI)⁺) The m/z calculation was 555.2005, finding a value of 555.1987.

N- (9- ((3aR,4R, 6aR) -6- (((3- (6- (hydroxyimino) methyl) pyridin-2-yl) prop-2-yn-1-yl) oxy) methyl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolan-4-yl) -9H-purin-6-yl) benzamide 60:

and

6- (3- (((3aR,4R,6R,6aR) -6- (6-amino-9H-purin-9-yl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolan-4-yl) methoxy) prop-1-yn-1-yl) pyridinaldoxime 61:

pyridine aldehyde 59(200mg, 0.361mmol, 1 equiv.), hydroxylamine hydrochloride (50mg, 0.722mmol, 2 equiv.) and CH₃CO₂A solution of Na (89mg, 1.083mmol, 3 equivalents) in dry ethanol (10mL) was stirred at reflux for 16 hours. After completion, the reaction mixture was concentrated under reduced pressure and the residue was purified by column chromatography eluting first with DCM to MeOH/DCM (2:98) to give product 60 as a white solid (80mg, 39%). IR (near Infrared) v_max 3196,2924,1698,1610,1581,1453,1246,1210,1075,907,727,644,551cm^-1；¹H NMR(400MHz,CDCl₃)δ(ppm)8.83(s,1H),8.38(s,1H),8.11(s,1H),7.99-7.89(m,2H),7.54-7.27(m,6H),6.27(d,J＝2.3Hz,2H),5.31(dd,J＝2.3,6.0Hz,1H),5.03(dd,J＝2.3,6.1Hz,1H),4.48(br q,J＝3.4Hz,1H),4.37,4.30(2d,J＝16.1Hz,2H),3.88(dd,J＝3.4,10.3Hz,1H),3.76(dd,J＝4.0,10.3Hz,1H),1.62(s,3H),1.38(s,3H)；¹³C NMR(100MHz,CDCl₃)δ(ppm)165.11,152.73,152.22,151.35,149.67,149.37,141.87,136.74,133.45,132.69,128.63,127.92,127.22,122.87,120.30,114.28,91.87,86.05,85.03,84.34,81.89,70.23,59.17,27.11,25.26；C₂₉H₂₈N₇O₆ ⁺HRMS (ESI)⁺) The m/z calculation was 570.2075, and the value was found to be 570.2096.

Further elution (MeOH/DCM 5:95) gave product 61(75mg, 45%) as a white solid. IR (near Infrared) v_max 3176,2925,1639,1450,1374,1207,1077,978,865,796,717,648,510cm^-1；¹H NMR(400MHz,CDCl₃)δ(ppm)8.30(s,1H),8.21(s,1H),8.15(s,1H),7.64-7.52(m,2H),7.26(s,1H),7.01-6.87(m,2H),6.20(d,J＝1.8Hz,2H),5.35(m,1H),5.03(m,1H),4.57(m,1H),4.35(m,2H),3.88-3.76(m,2H),1.61(s,3H),1.38(s,3H)；¹³C NMR(100MHz,CDCl₃)δ(ppm)155.35,152.55,152.47,149.34,149.08,141.96,139.32,136.75,132.82,127.10,119.93,114.13,91.70,86.10,85.72,84.86,84.38,81.88,70.21,59.11,27.07,25.27；C₂₂H₂₄N₇O₅ ⁺HRMS (ESI)⁺) The m/z calculation was 466.1812, and the value was found to be 466.1833.

Reference to the literature

Silvia, F. et al, J.Med.chem.2015, 58 th, 8269-8284

Synthesis of IV-bifunctional 3-methoxypyridine oxime analogs

Synthesis of 3-methoxy-6- (5-phenylpentyl) pyridylaldoxime 65:

3-methoxy-6- (5-phenylpent-1-yn-1-yl) pyridinecarboxaldehyde 63:

to commercially available 6-bromo-3-methoxypyridinal 62(75mg, 0.347mmol, 1.0 equiv.) in THF/Et₃Pd [ PPh ] was added to the degassed N (4mL/2mL)₃]₄(60mg, 0.052mmol, 0.15 equiv.) and CuI (20mg, 0.104mmol, 0.3 equiv.). After degassing the reaction mixture at room temperature for 5 minutes, alkyne 3(50mg, 0.347mmol, 1 eq) was added dropwise and the reaction mixture was stirred at room temperature for 16 hours. Upon completion (monitored by TLC), the reaction mixture was concentrated under reduced pressure and the residue was purified by column chromatography (EtOAc/PE 1:4) to afford the desired coupled methoxypyridinal 63(80mg, 83%) as a colorless liquid. R_f(30% EtOAc + PE) 0.25; IR (near Infrared) v_max 2941,2230,1709,1552,1466,1267,1007,747,699,542cm^-1；¹H NMR(400MHz,CDCl₃)δ(ppm)10.21(s,1H,H₁₈),7.52(d,J＝8.8Hz,1H,H₄),7.33(d,J＝8.8Hz,1H,H₅),7.28-7.13(m,5H,H₁₃-H₁₇),3.93(s,3H,-OMe),2.74(t,J＝7.5Hz,2H,H₁₁),2.39(t,J＝7.1Hz,2H,H₉),1.91(quintet,J＝7.1,7.5Hz,2H,H₁₀)；¹³C NMR(100MHz,CDCl₃)δ(ppm)195.52(C18),156.25,141.28,140.73,136.02,132.0,128.44,128.32,125.91,120.41(Ar),90.27(C7),79.44(C8),56.03(-OMe),34.86(C11),29.76(C10),18.72(C9)；C₁₈H₁₈N₁O₂ ⁺HRMS (ESI)⁺) The m/z calculation was 280.1332, and the value was found to be 280.1348.

3-methoxy-6- (5-phenylpent-1-yn-1-yl) pyridine aldoxime 64:

aldehyde 63(45mg, 0.161mmol, 1 equiv.), hydroxylamine hydrochloride (22mg, 0.322mmol, 2 equiv.) and CH₃CO₂A solution of Na (40mg, 0.483mmol, 3 equiv.) in dry ethanol (3mL) was stirred at reflux for 16 h. Upon completion (monitored by TLC), the solid was removed by filtration through a short pad of celite, the solvent was evaporated under reduced pressure and the residue was purified by column chromatography (EtOAc/PE 3:7) to afford oxime 64(45mg, 95%) as a white solid. R_f(50% EtOAc + PE) 0.35; IR (near Infrared) v_max 3247,2938,2234,1564,1463,1263,975,828,745,698,649,487cm^-1；^*1H NMR(400MHz,CDCl₃)δ(ppm)10.42(br s,1H,OH),8.42,8.10(2s,1.2H,H₁₈,H_18'),7.46-7.19(m,7.6H,Ar),3.94,3.91(2s,3.6H,-OMe),2.81,2.80(2t,J＝7.5Hz,2.4H,H₁₁,H_11'),2.47,2.44(2t,J＝7.1Hz,2.4H,H₉,H_9'),2.02-1.92(m,2.4H,H₁₀,H_10')；*¹³C NMR(100MHz,CDCl₃) Delta (. delta.) (ppm)153.54,151.66,147.87,141.41,141.17,140.65,140.23,136.37,135.53,131.86,129.18,128.47,128.34,128.29,127.83,125.93,125.84,119.29,118.69(Ar),90.98,89.24(C7),79.93,78.76(C8),55.93,55.70(-OMe),34.85,34.76(C11),29.86,29.73(C10),18.74,18.57(C9) (cis-to trans-isomer ratio of 1:5 only)^*)；C₁₈H₁₈N₂NaO₂ ⁺HRMS (ESI)⁺) The m/z calculation was 317.1260, and the value was found to be 317.1256.

3-methoxy-6- (5-phenylpentyl) pyridine aldoxime 65:

10% Pd/C (4.5mg, 0.042mmol, 0.5 equiv.) was added to a degassed solution of methoxypyridine oxime 64(25mg, 0.085mmol, 1 equiv.) in dry EtOAc (2 mL). By H₂After three washes, at room temperature, in H₂The reaction mixture was stirred for 3 hours (1 atm). Upon completion (monitored by TLC), the catalyst was removed by short column celite filtration, the solvent was evaporated, and the residue was purified by column chromatography (EtOAc/PE 1:9) to afford oxime 65(24mg, 95%) as a colorless liquid; r_f(50% EtOAc + PE) 0.40; IR (near Infrared) v_max 3253,2927,2855,1570,1464,1269,1127,975,746,698cm^-1；^*1H NMR(400MHz,CDCl₃)δ(ppm)8.40,8.02(2s,1.2H,H₁₈,H_18'),7.24-6.96(m,9.4H,Ar),3.79,3.78(2s,3.6H,-OMe),2.70-2.62(m,2.5H,H₁₁,H_11'),2.55-2.49(m,2.5H,H₇,H_7'),1.68-1.52(m,5H,H₈,H_8',H₁₀,H_10'),1.35-1.28(m,2.5H,H₉,H_9')；*¹³C NMR(100MHz,CDCl₃) δ (ppm)154.25,152.57,150.87,150.67,146.82,142.71,142.47,140.12,139.24,137.09,128.35,128.21,128.16,125.59,125.51,125.06,123.58,119.91,119.18(Ar),55.82,55.63(-OMe),37.23,36.25(C11),35.78,35.73(C7),31.25,31.12(C10),29.85,29.67(C9),28.90,28.65(C8) (. cis-trans-isomer to trans-isomer ratio of 1: 4); c₁₈H₂₃N₂O₂ ⁺HRMS (ESI)⁺) The m/z calculation was 299.1754, and the value was found to be 299.1740.

Synthesis of V-quinoline derived methoxypyridine oxime analogs：

3-methoxy-6- (4- (quinolin-4-ylamino) but-1-yn-1-yl) pyridylaldehyde 66:

to a commercial 6-bromo-3-methoxypyridinal 62(97mg, 0.448mmol, 1.1 equiv.) in THF/Et₃Adding Pd [ PPh ] into N (3mL/3mL) degassed solution₃]₄(71mg, 0.061mmol, 0.15 equiv.) and CuI (23mg, 0.122mmol, 0.3 equiv.). After degassing the reaction mixture at room temperature for 5 minutes, alkyne 24(80mg, 0.408mmol, 1 eq) in THF (3mL) was added dropwise and the reaction mixture was stirred at room temperature for 16 hours. Upon completion (monitored by TLC), the reaction mixture was concentrated under reduced pressure and the residue was purified by column chromatography (MeOH/EtOAc 1:9) to afford the desired coupled methoxypyridinal 66(126mg, 93%) as a light yellow solid. R_f(30% MeOH + EtOAc) 0.25; IR (near Infrared) v_max3281,2926,2233,1704,1582,1434,1267,1126,1009,763,694,521,494cm^-1；¹H NMR(400MHz,CDCl₃)δ(ppm)10.21(s,1H,H₁₈),7.52(d,J＝8.8Hz,1H,H₄),7.33(d,J＝8.8Hz,1H,H₅),7.28-7.13(m,5H,H₁₃-H₁₇),3.93(s,3H,-OMe),2.74(t,J＝7.5Hz,2H,H₁₁),2.39(t,J＝7.1Hz,2H,H₉),1.91(quintet,J＝7.1,7.5Hz,2H,H₁₀)；¹³C NMR(100MHz,CDCl₃)δ(ppm)195.52(C18),156.25,141.28,140.73,136.02,132.0,128.44,128.32,125.91,120.41(Ar),90.27(C7),79.44(C8),56.03(-OMe),34.86(C11),29.76(C10),18.72(C9)；C₁₈H₁₈N₁O₂ ⁺HRMS (ESI)⁺) The m/z calculation was 280.1332, and the value was found to be 280.1348.

3-methoxy-6- (4- (quinolin-4-ylamino) but-1-yn-1-yl) pyridine aldoxime 67:

aldehyde 66(100mg, 0.362mmol, 1 equiv.), hydroxylamine hydrochloride (50mg, 0.724mmol, 2 equiv.) and CH₃CO₂A solution of Na (89mg, 1.086mmol, 3 equiv.) in dry ethanol (5mL) was stirred at reflux for 16 h. After completion (by T)LC monitoring), the solid was removed by filtration through a short pad of celite, the solvent was evaporated under reduced pressure, and the residue was purified by column chromatography (MeOH/EtOAc 1:9) to afford oxime 67(65mg, 62%) as a white solid. R_f(30% MeOH + EtOAc) 0.2; IR (near Infrared) v_max 3319,2924,1897,1586,1460,1242,1115,982,829,760,649,524cm^-1；¹H NMR(500MHz,DMSO-d6)δ(ppm)11.63(br s,1H,OH),8.41(d,J＝5.2Hz,1H,Ar),8.22(s,1H,-C-NOH),8.21(d,J＝8.5Hz,1H,Ar),7.79(d,J＝8.5Hz,1H,Ar),7.61(t,J＝7.6Hz,1H,Ar),7.50-7.36(m,4H,Ar),6.58(d,J＝5.4Hz,1H,Ar),3.85(s,3H,-OMe),3.58(q,J＝5.9,7.1Hz,2H,H₁₀),2.83(t,J＝7.1Hz,2H,H₉)；¹³C NMR(125MHz,DMSO-d6)δ(ppm)153.77,151.22,149.97,148.83,145.06,140.87,134.59,129.57,129.24,128.54,124.42,122.07,120.24,119.30,98.94,(Ar),87.06(C8),81.50(C7),56.46(-OMe),41.75(C10),19.08(C9)；C₂₀H₁₉N₄O₂ ⁺HRMS (ESI)⁺) The m/z calculation was 347.1503, and the value was found to be 347.1491.

4- ((4- (6- ((hydroxyimino) methyl) -5-methoxypyridin-2-yl) but-3-yn-1-yl) amino) quinoline-1-ammonium chloride 68:

to MeOH/H₂To compound 67(9.5mg) in O (0.5mL/0.5mL) was added 1.2N HCl (0.1mL) and stirred for 2 minutes, and allowed to stand at room temperature for 10 minutes. The reaction mixture was concentrated under reduced pressure to provide HCl salt 68 as a white solid in quantitative yield. IR (near Infrared) v_max 3186,3099,2838,2237,1615,1593,1449,1277,1007,760,649,530,491cm^-1；¹H NMR(500MHz,D₂O)δ(ppm)*8.18-8.08(m,3H,Ar),*7.96-7.91(m,1.5H,Ar),*7.74-7.70(m,1.5H,Ar),*7.62-7.54(m,3H,Ar),*7.51-7.40(m,4.5H,Ar),*6.73-6.70(m,1.5H,Ar),*3.92(s,1.5H,-OMe),*3.84(t,J＝6.6Hz,1H,H_10'),3.80(s,3H,-OMe),3.75(t,J＝6.6Hz,2H,H₁₀),*2.98(t,J＝6.6Hz,1H,H_9'),2.89(t,J＝6.6Hz,2H,H₉)；*¹³C NMR(125MHz,D₂O)δ(ppm)156.49,154.95,142.15,142.03,141.86,141.45,140.04,139.68,138.68,137.60,137.00,134.19,131.06,130.37,128.51,128.26,17.55,127.52,125.38,122.39,120.29,120.22,119.12,116.93,116.81,98.55,98.47(Ar),94.74(C8),76.76(C7),57.71,57.22(-OMe),41.65,41.10(C10),19.5(C9)(^*1:2 of cis-trans isomers); c₂₀H₂₀ClN₄O₂ ⁺HRMS (ESI)⁺) The m/z calculation was 347.1503, and the value was found to be 347.1461.

3-methoxy-6- (4- (quinolin-4-ylamino) butyl) pyridine aldoxime 69:

10% Pd/C (4.5mg, 0.042mmol, 0.5 equiv.) was added to a degassed solution of methoxypyridine oxime 67(25mg, 0.085mmol, 1 equiv.) in dry EtOAc (2 mL). By H₂After three washes, at room temperature, in H₂The reaction mixture was stirred for 3 hours (1 atm). Upon completion (monitored by TLC), the catalyst was removed by short column celite filtration, the solvent was evaporated under reduced pressure, and the residue was purified by column chromatography (EtOAc/PE 1:9) to afford oxime 69(24mg, 95%) as a colourless liquid; r_f(50% EtOAc + PE) 0.40; IR (near Infrared) v_max 3327,2923,2853,1582,1457,1272,1126,968,763,694,540,473cm^-1；^*1H NMR(400MHz,CDCl₃)δ(ppm)8.42(2s,1H,H₁₈,H_18'),8.32(d,J＝5.7Hz,1H,Ar),8.10(dd,J＝1.2,8.6Hz,1H,Ar),7.79(d,J＝1.2,8.6Hz,1H,Ar),7.64(m,1H,Ar),7.44(m,1H,Ar),7.39(d,J＝8.7Hz,1H,Ar),7.26(d,J＝8.7Hz,1H,Ar),6.50(d,J＝5.8Hz,1H,Ar),3.87(s,3H,-OMe),3.42(t,J＝6.9Hz,1H,H₁₀),3.42(t,J＝6.9Hz,1H,H₁₀),2.83(t,J＝7.5Hz,1H,H₇),1.87-1.77(m,4H,H₈,H₉)；*¹³C NMR(100MHz,CDCl₃)δ(ppm)155.21,154.35,153.13,150.73,148.33,145.93,140.73,138.63,130.85,128.33,125.82,122.51,121.43,120.31,99.30(Ar),56.53(-OMe),43.86(C10),37.37(C7),28.95(C9),28.82(C8)(^*Cis-trans isomer ratio of 1: 4): c₂₀H₂₃N₄O₂ ⁺The calculated result of HRMS (ESI +) m/z of (D) was 351.1816, and the found value was 351.1827.

4- ((4- (6- ((hydroxyimino) methyl) -5-methoxypyridin-2-yl) butyl) amino) quinolin-1-amine 70:

to compound 69(8mg) in methanol/water (0.5mL/0.5mL) was added 1.2N HCl (0.1mL) and stirred for 2 minutes, and allowed to stand at room temperature for 10 minutes. The reaction mixture was concentrated under reduced pressure to obtain HCl salt 70 as a white solid in quantitative yield. IR (near Infrared) v_max 3237,3111,2926,1617,1594,1452,1291,1011,764,663,592cm^-1；¹H NMR(500MHz,D₂O)δ(ppm)8.20(s,1H,H₁₈),8.19(d,J＝8.6Hz,1H,Ar),8.04(d,J＝8.6Hz,1H,Ar),7.94-7.87(m,2H,Ar),7.75(dd,J＝8.6,18.8Hz,1H,Ar),7.64(t,J＝8.8Hz,1H,Ar),6.64(d,J＝7.2Hz,1H,Ar),3.93(s,3H,-OMe),3.55(t,J＝6.5Hz,1H,H₁₀),3.01(t,J＝6.8Hz,1H,H₇),1.95-1.80(m,4H,H₈,H₉)；*¹³C NMR(125MHz,D₂O) δ (ppm)156.07,154.21,150.14,141.63,139.57,137.60,134.14,133.86,129.10,128.69,127.47,122.29,120.27,116.86,98.31(Ar),57.45(-OMe),42.83(C10),32.20(C7),26.12(C9),25.81(C8) (cis/trans ratio 1: 2); c₂₀H₂₃N₄O₂ ⁺The calculated HRMS (ESI +) m/z of (D) was 351.1816, and found to be 351.1782.

(3aS,4S,6R,6aR) -6- (6-amino-9H-purin-9-yl) -N- (4- (6- ((hydroxyimino) methyl) -5-methoxypyridin-2-yl) but-3-yn-1-yl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolane-4-carboxamide 72:

to the commercially available 6-bromo-3-methylOxopyridinal 62(192mg, 0.887mmol, 1.1 equiv.) in THF/Et₃Pd [ PPh ] was added to a degassed solution in N (5mL/5mL)₃]₄(140mg, 0.121mmol, 0.15 equiv.) and CuI (46mg, 0.242mmol, 0.3 equiv.). After degassing the reaction mixture at room temperature for 5 minutes, alkyne 54(300mg, 0.806mmol, 1 eq) in THF was added dropwise (5mL) and the reaction mixture was stirred at room temperature for 16 hours. After completion (monitored by TLC), the reaction mixture was concentrated under reduced pressure and the residue was passed through a mini-filter column (MeOH/EtOAc 5:95) to afford the desired coupled methoxypyridinal 71 as a light yellow solid (360mg, 88%). This crude aldehyde was used in the next step without purification.

Aldehyde 71(220mg, 0.433mmol, 1 equiv.), hydroxylamine hydrochloride (60mg, 0.867mmol, 2 equiv.) and CH₃CO₂A solution of Na (107mg, 1.299mmol, 3 equiv.) in dry ethanol (7mL) was stirred at reflux for 16 h. Upon completion (monitored by TLC), the solid was removed by filtration through a short pad of celite, the solvent was evaporated, and the residue was purified by column chromatography (MeOH/EtOAc 1:9) to provide oxime 72(180mg, 78%) as a white solid. R_f(30% MeOH + EtOAc) 0.2; IR (near Infrared) v_max 3185,2926,1640,1464,1264,1209,1090,971,868,797,647,510cm^-1；¹H NMR(400MHz,MeOD)δ(ppm)8.39-8.11(3s,3H,Ar,-C＝NOH),7.37(d,J＝8.6,1H,H₄),7.26(d,J＝8.6Hz,1H,H₅),6.34(br s,H,-CH),5.57(dd,J＝1.8,6.0Hz,1H,-CH),5.41(br d,J＝6.0Hz,1H,-CH),4.68(d,J＝1.8Hz,1H,-CH),3.89(s,3H,-OCH₃),3.21(m,1H,-CH₂),3.09(m,1H,-CH₂),2.27(m,1H,-CH₂),2.10(m,1H,-CH₂),1.57(s,3H,-CH₃),1.37(s,3H,-CH₃)；¹³C NMR(100MHz,MeOD)δ(ppm)172.07,157.33,155.15,153.99,150.30,145.21,142.55,141.69,135.92,129.89,120.84,120.49,115.17(Ar),92.41(C16),88.65(C15),87.34(C8),85.36(C14),85.26(C13),81.58(C7),56.74(C30),38.79(C10),27.29(C28),25.54(C29),20.47(C9)；C₂₄H₂₇N₈O₆ ⁺HRMS (ESI)⁺) The m/z calculation was 523.2048, and the value was found to be 523.2038.

Synthesis of VI-trifunctional ethylcarbazole compound

6- (azidomethyl) pyridylaldehyde 75:

synthesis of methyl 6- (azidomethyl) picolinate 74 by Harekrushne, B¹Is achieved by well established procedures.

DIBAL-H (1M in CH) at-78 deg.C₂Cl₂1.563mL, 1.563mmol, 3 equiv) was added dropwise to azido ester 74(100mg 0.521mmol, 1 equiv) in dry CH₂Cl₂(5mL) and the reaction mixture was stirred at-78 deg.C for 5 hours. After completion of the reaction, the reaction mixture was quenched with methanol (3mL) and the cooling tank was removed. When the mixture is warmed to room temperature, H is used₂The reaction mixture was diluted with EtOAc and extracted. The combined organic layers were washed with MgSO 2₄And drying. The solids were filtered off and the solvent was evaporated to give the aldehyde 75. The crude aldehyde 75 was used directly in the next step without purification. IR (near Infrared) v_max2836,2098,1709,1591,1457,1255,990,777,641cm^-1；¹H NMR(400MHz,CDCl₃)δ(ppm)10.04(s,1H),7.94-7.86(m,2H),7.56(m,2H),4.57(s,2H)；¹³C NMR(100MHz,CDCl₃)δ(ppm)193.07,156.73,152.59,138.11,126.05,102.78,55.15；C₇H₇N₄O₁ ⁺HRMS (ESI)⁺) The calculated m/z was 163.0604, which was found to be 163.0614.

6- (azidomethyl) pyridine aldoxime 76:

crude pyridylaldehyde 75 (0)521mmol, 1 eq), hydroxylamine hydrochloride (73mg, 1.042, 2 eq) and CH₃CO₂A solution of Na (128mg, 1.563mmol, 3 equiv.) in dry ethanol (5mL) was stirred at 80 ℃ for 16 h. Upon completion, the solids were removed by short column celite filtration, the solvent was evaporated, and the residue was purified by column chromatography (EtOAc/P.E:1:9) to provide oxime 76(70mg, 76%) as a thick syrup. IR (near Infrared) v_max 3182,3010,2889,2084,1572,1590,1459,12666,1233,1158,994,966,782,741,651,619,501cm^-1；¹H NMR(400MHz,CD₃OD)δ(ppm)8.10(s,1H),7.86-7.76(m,2H),7.40(dd,J＝1.8,6.8Hz,2H),4.48(s,2H)；¹³C NMR(100MHz,CD₃OD)δ(ppm)157.29,153.94,149.93,139.26,1323.66,120.86,56.16；C₇H₈N₅O₁ ⁺HRMS (ESI)⁺) The calculated m/z was 178.0721, which was found to be 178.0723508.1867.

(3aS,4S,6R,6aR) -6- (6-amino-9H-purin-9-yl) -N- (2- (1- ((6- ((hydroxyimino) methyl) pyridin-2-yl) methyl) -1H-1,2, 3-triazol-4-yl) ethyl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolane-4-carboxamide 77:

to t-BuOH/H₂To a stirred solution of oxime 76(52mg, 0.295mmol, 1.1 equiv) in O (2mL/1.5mL) was added CuSO₄(17mg, 0.107mmol, 0.4 equiv.), sodium ascorbate (21mg, 0.107mmol, 0.4 equiv.), and alkyne 54(100mg, 0.268mmol, 1 equiv.). The reaction mixture was allowed to stir at 80 ℃ for 6 hours. After completion (monitored by TLC), the reaction mixture was concentrated under reduced pressure and the residue was purified by column chromatography (MeOH/EtOAc: 5:95) to provide the desired triazole compound 77(85mg, 58%) as a white solid. IR (near Infrared) v_max 3192,2924,1644,1598,1458,1376,1209,1155,1057,992,868,797,648,511cm^-1；¹H NMR(400MHz,CD₃OD)δ(ppm)8.20(s,1H),8.08(s,1H),8.05(s,1H),7.75-7.70(m,2H),7.12(m,1H),6.30(d,J＝1.7Hz,1H),5.62(s,2H),5.50(dd,J＝1.9,6.1Hz,1H),4.62(d,J＝1.9Hz,1H),3.24-3.01(m,2H),2.61-2.41(m,2H),1.57(s,3H),1.38(s,3H)；¹³C NMR(100MHz,CD₃OD)δ(ppm)171.91,157.38,156.08,153.97,150.27,150.06,146.45,142.54,139.28,124.46,123.53,121.07,120.55,115.24,92.65,88.38,85.28,85.06,56.09,39.64,27.31,25.91,25.55；C₂₄H₂₈N₁₁O₅ ⁺HRMS (ESI)⁺) The calculated m/z was 550.2237, which was found to be 550.2269.

(2S,3S,4R,5R) -5- (6-amino-9H-purin-9-yl) -3, 4-dihydroxy-N- (2- (1- ((6- ((hydroxyimino) -methyl) pyridin-2-yl) methyl) -1H-1,2, 3-triazol-4-yl) ethyl) tetrahydrofuran-2-carboxamide hydrochloride 78:

to a stirred solution of triazole 77(23mg, 0.042mmol, 1 equiv) in dry methanol (2mL) was added 1.2N HCl (127 μ L, 0.42mmol, 10 equiv) and the reaction mixture was stirred at 55 ℃ for 4 h. After completion, the reaction mixture was directly concentrated under reduced pressure and purified by reverse phase column chromatography (MeOH/H)₂O3:7) to provide HCl salt 78(15mg, 72%) as a white solid. IR (near Infrared) v_max 3196,1640,1588,1458,1427,1306,1254,1113,1054,997,796,647cm^-1；¹H NMR(400MHz,CD₃OD)δ(ppm)7.99(s,1H),8.89(s,1H),7.83(s,1H),7.42(t,J＝7.8Hz,1H),7.29(t,J＝9.3Hz,1H),7.20(s,1H),7.13(t,J＝7.8Hz,1H),5.77(d,J＝8.3Hz,1H),5.44-5.38(2d,J＝14.8Hz,2H),4.43(s,1H),4.32(br d,J＝5.0Hz,1H),4.08(dd,J＝4.8,8.3Hz,1H),3.61(m,1H),3.35(m,1H),3.08-299(m,1H),3.96-288(m,1H)；¹³C NMR(100MHz,CD₃OD)δ(ppm)172.32,155.44,153.78,151.99,150.84,147.61,142.13,138.97,124.62,124.41,121.56,119.75,89.21,85.20,73.72,72.06,55.28,49.50(MeOH),39.92,24.77；C₂₁H₃₃₄N₁₁O₅ ⁺HRMS (ESI)⁺) The m/z calculation was 510.1957, and the value was found to be 510.1956.

Reference to the literature

Harekrushna B. et al, chem. Eur. J.2015, 21 st, 10179-p 10184.

Synthesis of VII-trifunctional Pseudoethylcarbazole Compounds

N- (9- ((3aR,4R, 6aR) -6- (((1- ((6- ((hydroxyimino) methyl) pyridin-2-yl) -1H-1,2, 3-triazol-4-yl) methoxy) methyl) -2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolan-4-yl) -9H-purin-6-ylbenzamide 79:

to t-BuOH/H₂To a stirred solution of oxime 76(44mg, 0.245mmol, 1.1 equiv) in O (2mL/1.5mL) was added CuSO₄(17mg, 0.045mmol, 0.2 equiv.), sodium carbonate (18mg, 0.045mmol, 0.2 equiv.), and alkyne 58(100mg, 0.223mmol, 1 equiv.). The reaction mixture was allowed to stir at 80 ℃ for 6 hours. After completion (monitored by TLC), the reaction mixture was concentrated under reduced pressure and the residue was purified by column chromatography (pure EtOAc-MeOH/EtOAc: 5:95) to provide the desired triazole compound 79(100mg, 72%) as a white solid. IR (near Infrared) v_max 2924,1698,1610,1581,1455,1248,1211,1070,994,796,709,645,563cm^-1；¹H NMR(400MHz,CD₃OD)δ(ppm)10.88(br s,1H),9.31(br s,1H),8.77(s,1H),8.31(s,1H),8.14(s,1H),7.95-7.80(m,3H),7.51-7.28(m,5H),6.99(d,J＝7.5Hz,1H),6.25(d,J＝2.3Hz,1H),5.63(s,2H),5.21(dd,J＝2.2,5.9Hz,1H),4.98(dd,J＝1.6,5.9Hz,1H),4.65-4.50(m,3H),3.82(dd,J＝2.2,10.6Hz,1H),3.70(dd,J＝3.0,10.6Hz,1H),1.61(s,3H),1.37(s,3H)；¹³C NMR(100MHz,CD₃OD)δ(ppm)165.35,154.38,152.69,151.64,151.44,150.02,149.19,143.91,141.90,137.62,133.31,128.56,127.93,124.13,122.78,122.07,120.42,114.09,92.26,86.27,85.28,81.85,70.67,64.47,55.03,27.11,25.24；C₃₀H₃₁N₁₀O₆ ⁺HRMS (ESI)⁺) The m/z calculation was 627.2423.

Example 2: in vitro reactivation of human acetylcholinesterase (hAChE) by compounds of the invention

Compounds 57, 78, 25 and 26 of example 1 were tested to determine their activation performance on hAChE inhibited by O-ethyl S- [2- (diisopropylamino) ethyl ] methyl phosphorothioate (VX), taben, sarin or paraoxon.

2-PAM (pralidoxime or 2- [ (E) - (hydroxyimino) methyl ] -1-methylpyridine) and HI6 (doxime chloride or [1- [ (4-carbamoylpyridin-1-am-1-yl) methoxymethyl ] pyridin-2-ylidene ] methyloxazadichloride) were used as comparative compounds.

The medical treatment scheme is as follows:

materials and methods have been described in WO2017021319, 2014 journal of european medicines chemistry 2014 78,455-467 and 2018 journal of medical chemistry 61, 7630-7639.

IC 50. Recombinant hAChE produced and purified as described previously (Carletti et al, 2008, J Am Chem Soc 130 (47): 1601-20). Compounds were dissolved in methanol to form 5mM or 10mM stock solutions and further diluted in phosphate buffer (sodium phosphate 0.1M, pH 7.4). The recombinant hAChE activity was measured spectrophotometrically (absorbance at 412 nm) in the presence of different concentrations of oxime in 1mL of Ellman buffer (sodium phosphate 0.1M, pH 7.4, 0.1% BSA, 0.5mM DTNB, 25 ℃). At least two measurements were made for each test concentration. Using profit (quantumsoft) nonlinear fitting, using standard IC50 equations: % activity ═ 100 IC₅₀/(IC₅₀+[Ox]) The concentration of compound that produced 50% enzyme inhibition was determined.

Inhibition of hAChE by OPNAs. Recombinant hAChE was produced and purified as described previously (see reference: http:// www.ncbi.nlm.nih.gov/pubmed/18975951). VX and Taben are from DGA maitrise NRBC (Vert le Petit, France). Stock solutions of VX, sarin, taber and paraoxon were 5mM in isopropanol. The inhibition of 120M hAChE was tested with a 5-fold excess of OPNAs and in Tris buffer (20mM, pH 7.4, 0.1% BSA) at 25 ℃. After 20 min incubation, the inhibited hAChE was desalted on a PD-10 column (GE Healthcare).

OPNAs inhibit the reactivation of hAChE. The OPNA inhibited hAChE was incubated with oxime at a concentration of at least 4 or 5 in phosphate buffer (0.1M, pH 7.4, 0.1% BSA) at 37 ℃. The final concentration of methanol in the mixed broth was less than 2%, which had no effect on the stability of the enzyme. At time intervals of 1-10 minutes (depending on the reactivation rate), 10 aliquots of each solution containing different concentrations of oxime were transferred to tubes containing 1mM acetylcholine and placed in 1mL of Ellman buffer (phosphate 0.1M, pH 7.4, 0.1% BSA, 0.5mM DTNB, 25 ℃) to measure the activity of hAChE.

The enzyme activity of the control group remained unchanged during the experiment. Percentage of reactivated enzyme (% E)_{Reaction of}) Calculated as the ratio of recovered enzyme activity to enzyme activity in the control. The apparent reactivation ratio kobs, dissociation constant K for the inhibitory enzyme oxime complex (E-POx) for each oxime concentration was calculated by nonlinear fitting (Quantum Soft) with ProFit using the standard oxime concentration dependent reactivation equation derived from the following protocol_DAnd the maximum reactivation ratio constant k_r：

E-P ═ enzyme

Oxime compounds

E-Pox ═ enzyme oxime complexes

The test results are shown below (tables 1 and 2):

table 1 shows the human hAChE reactivation performance on OP inhibition by oximes 2-PAM, HI6, 57, 78, 25 and 26

Table 1 shows: compound 57 exhibits greater potency against VX, Talbot, sarin and paraoxon than the reference compounds 2-PAM and HI6High reactivation kinetics (k)_r，min^-1) (ii) a Compound 78 shows higher reactivation kinetics for VX, taber and sarin (k) compared to the reference compounds 2-PAM and HI6 (k)_r，min^-1)。

Compounds 25 and 26 show strong affinity for VX or sarin inhibited AChE. These compounds showed higher reactivities than 2-PAM and HI6 (kr2 mM)^-1min^-1)。

Table 2 is an oxime compound: IC of 2-PAM, HI6, 57, 78, 25 and 26 on AChE₅₀

Table 2 shows that: compounds 25 and 26 showed very high affinity for AChE, which is higher than that of 2-PAM and HI 6. Compound 25 exhibited the best affinity.