A palmatine derivative, a preparation method and application thereof

Abstract

The present application provides a kind of palmatine derivative, the structural general formula is as follows: the present application selects palmatine as raw material, through multi-step synthesis, corresponding compound with high yield can be obtained, the palmatine derivative can effectively resist antibiotic resistant helicobacter pylori strain, especially spherical mutant strain, has good bacteriostatic activity, solves the antibiotic resistance and retention recurrence problem of helicobacter pylori in clinical treatment, is suitable for industrial production and clinical application.

Description

Technical Field

[0002] This invention relates to the field of pharmaceutical technology, specifically to a palmatine derivative, its preparation method, and its application. Background Technology

[0004] Helicobacter pylori is a pathogenic bacterium that can colonize the highly acidic environment of the human stomach. The International Agency for Research on Cancer (IARC) classifies it as a Group 1 carcinogen and a major contributing factor to chronic gastritis, peptic ulcers, and even gastric cancer. Currently, clinical eradication of Helicobacter pylori mainly relies on combination therapy (triple or quadruple therapy) containing two or more antibiotics (such as clarithromycin, metronidazole, amoxicillin, and levofloxacin). However, due to the widespread use of antibiotics, drug resistance in Helicobacter pylori is becoming increasingly serious. Clarithromycin and metronidazole, as first-line treatments, have seen a continuous increase in resistance rates globally, becoming a major cause of eradication treatment failure. The development of drug resistance forces clinicians to extend treatment duration or change treatment regimens, but this often leads to decreased patient compliance and increased adverse drug reactions, creating a vicious cycle. Therefore, developing novel anti-Helicobacter pylori drugs with entirely new mechanisms of action that can effectively overcome existing drug resistance has become a pressing technical challenge in this field.

[0005] Existing technology

[0006] Berberine is a commonly used clinical drug, belonging to the isoquinoline alkaloid class. Its English name comes from the plant Fibraurea in the palm family. This substance is widely found in medicinal plants such as Coptis chinensis. It has structural characteristics similar to berberine and berberine, and possesses antibacterial activity.

[0007]

[0008] Isoquinoline alkaloids are a class of natural products widely found in the plant kingdom, exhibiting structural diversity and a wide range of biological activities. Existing literature reports various pharmacological effects, including antitumor, antibacterial, and anti-inflammatory properties. Specifically in the field of antibacterial activity, reviews have pointed out that alkaloids form the structural framework of important antibacterial drugs such as metronidazole and quinolones, and some isoquinoline alkaloids have shown the potential for direct antibacterial activity or enhancing antibiotic activity.

[0009] The applicant’s previous research found that the natural product palmatine with an isoquinoline ring has certain anti-Helicobacter pylori activity but weak antibacterial activity, with a minimum inhibitory concentration (MIC) of 60-300 μg / mL [1]. The applicant found that the antibacterial activity of monosubstituted palmatine derivatives (especially those with nitrogen-containing side chains) was improved compared with the original palmatine drug, with a minimum MIC of 4 μg / mL. The applicant further used purslane as raw material to synthesize a series of 3,13-disubstituted isoquinoline derivatives (ZL 202311367970.0 and ZL202510593023.6). The compounds were verified to have better anti-Helicobacter pylori activity, among which the four compounds 4a-4d in ZL 202311367970.0 had the best anti-Helicobacter pylori activity (the MIC of 4b was 0.25-2 μg / mL). The applicant continues to address bottlenecks in clinical drug use, exploring compounds with greater development and clinical translational value to lay the foundation for subsequent drug development iterations and adjust research directions and concepts. Research has found that Helicobacter pylori can form spherical bodies, an adaptive survival state for the bacterium in harsh environments (such as exposure to air or antibiotic treatment). This spherical transformation enhances its environmental tolerance and resistance, allowing it to survive for extended periods in water, creating favorable conditions for oral-oral and fecal-oral transmission. After spherical transformation, the bacteria's metabolic activity decreases, entering a dormant state that allows it to evade antibiotics. Once the environment becomes suitable, these spherical bodies can revert to metabolically active spiral shapes, leading to infection recurrence. Developing drugs that effectively kill spherical bacteria can provide new insights for clinical drug use. This study investigates whether newly synthesized compounds can exert antibacterial effects against both spiral and spherical resistant Helicobacter pylori, representing a novel strategy for the clinical use of palmatine derivatives, which has not been previously reported.

[0010] References:

[0011] [1] Fan T , Guo X , Zeng Q ,et al.Synthesis and Structure-ActivityRelationship of Palmatine Derivatives as a Novel Class of AntibacterialAgents against Helicobacter pylori[J]. Molecules, 2020, 25(6).DOI:10.3390 / molecules25061352. Summary of the Invention

[0013] To address the aforementioned technical issues, the applicant revised its research plan and concept around palmatine and synthesized a series of palmatine-like disubstituted derivatives. It was found that the newly synthesized palmatine derivatives had better antibacterial effects. Based on the above research results, the applicant proposed its own technical solution, namely, a palmatine derivative, wherein the palmatine derivative is a palmatine derivative with disubstituted palmatine at positions 9 and 13.

[0014] This invention differs from previous compounds in that it can replace the methyl group at the 9-position of the methoxy group in palmatine, significantly altering the properties of palmatine compounds. This is fundamentally different from previous palmatine derivatives produced by replacing the methyl group at the 3-position and the hydrogen at the 13-position with other groups.

[0015] Therefore, in this invention application, the general structural formula of the palmatine derivative is as follows:

[0016] ,

[0017] R1 is selected from any one of the C2-C6 alkenyl groups;

[0018] The R2 is selected from C3-C8 alkenyl, C3-C8 alkynyl, benzyl group substituted with one or two independently selected C1-C4 alkyl, C1-C4 alkoxy or phenyl or nitro groups, and naphthylmethyl.

[0019] Furthermore, in the above-mentioned barmatine derivatives, R1 is selected from isopentenyl, 2-butenyl.

[0020] Furthermore, in the above-mentioned palmatine derivatives, R2 is selected from the following substituents:

[0021] , ,

[0022] , ,

[0023] , ,

[0024] , ,

[0025] , ,

[0026] , ,

[0027] , ,

[0028] , and .

[0029] Furthermore, the palmatine derivative is selected from the following compounds:

[0030] , ,

[0031] , ,

[0032] , ,

[0033] , ,

[0034] , ,

[0035] , ,

[0036] , ,

[0037] , ,

[0038] , ,

[0039] , ,

[0040] , ,and .

[0041] The compounds mentioned above are the compounds in Examples 1-23.

[0042] The present invention provides a pharmaceutical composition comprising the above-mentioned palmatine derivative or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

[0043] This invention provides a method for preparing the above-mentioned palmatine derivative, wherein the preparation method comprises the following steps:

[0044] 1) Take palmatine hydrochloride, heat to remove the methyl group at the 9-position, add acid solution to acidify, and obtain a dark red solid;

[0045] 2) The dark red solid obtained in step 1) is added to an aprotic solvent, a catalyst and a 9-substituent starting material are added, and after heating to cause a 9-substitution reaction, the organic phase is concentrated and the residue is concentrated by column chromatography to obtain the nucleophilic substitution product.

[0046] 3) Add an aprotic solvent, a reducing agent and a catalyst to the substitution product in step 2), heat to activate, add the 13-substituent raw material to carry out the 13-substitution reaction, after the reaction is completed, cool until the solid is completely precipitated, filter, and purify the filtrate by chromatographic column to obtain the palmatine derivative.

[0047] The aprotic solvent in steps 2) and 3) is selected from anhydrous acetonitrile, tetrahydrofuran, and DMF. The reducing agent is a commonly used reducing agent in nucleophilic substitution reactions, such as potassium borohydride or sodium borohydride.

[0048] The catalyst is a commonly used catalyst in nucleophilic substitution reactions, such as sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, potassium tert-butoxide, and sodium hydride.

[0049] The selection of commonly used reducing agents and catalysts is well known to those skilled in the art, and relevant published literature, textbooks and reference books can be consulted.

[0050] The order of steps for the 9-position substitution reaction and the 13-position substitution reaction is interchangeable.

[0051] The acid solution is selected from common acids such as dilute hydrochloric acid, hydrochloric acid methanol, and hydrochloric acid ethanol, in order to protect the 9-OH group.

[0052] This invention further provides the use of the above-mentioned palmatine derivative in the preparation of a medicament for treating Helicobacter pylori infection. The Helicobacter pylori infection-related diseases include Helicobacter pylori infection, and functional dyspepsia, gastritis, peptic ulcers, gastric mucosa-associated lymphoid tissue lymphoma, and gastric cancer caused by Helicobacter pylori infection. The gastritis includes chronic active gastritis and chronic atrophic gastritis.

[0053] The 9-,13-disubstituted palmatine derivatives involved in this invention can effectively inhibit Helicobacter pylori strains resistant to clarithromycin and metronidazole. They are effective against clinically resistant strains with different resistance phenotypes, exhibiting superior activity compared to existing 3-,13-disubstituted isoquinoline drugs. They possess a novel mechanism of action different from existing antibiotics to avoid cross-resistance. Furthermore, this series of compounds is effective against antibiotic-induced cocci, with activity far exceeding that of 3-,13-disubstituted palmatine, thus expanding the therapeutic spectrum of Helicobacter pylori resistant bacteria and greatly meeting clinical needs. Attached Figure Description

[0055] Figure 1: Verification of growth inhibition diagram of cocci.

[0056] a) shows the growth of spherical Helicobacter pylori on the culture medium when the drug fails to inhibit the growth of spherical bacteria. b) shows the absence of Helicobacter pylori growth on the culture medium under the minimum inhibitory concentration of the drug. Detailed Implementation

[0058] The specific embodiments of the present invention are described below to enable those skilled in the art to understand the present invention. However, it should be understood that the present invention is not limited to the scope of the specific embodiments. For those skilled in the art, various changes are obvious as long as they are within the spirit and scope of the present invention as defined and determined by the appended claims. All inventions utilizing the concept of the present invention are protected.

[0059] Example 1 Synthesis of 13-((E)-2-buten-1-yl)-9-(((E)-2-buten-1-yl)oxy)-2,3,10-trimethoxy-5,6-dihydroisoquinolino[3,2-a]isoquinolino-7-onium (Compound 1)

[0060]

[0061] Parmaline hydrochloride (2.78 g, 7.5 mmol) was placed in a 250 mL round-bottom flask, and the mixture was heated to 195°C for 45 min under a negative pressure of 20–30 mmHg to obtain a deep red solid. The solid was acidified with ethanol / concentrated hydrochloric acid (95 mL, 5.0 mL) and evaporated to dryness to obtain a dark red compound. Potassium carbonate (0.41 g, 3.0 mmol) and crotonyl bromide (first reactant, 0.40 g, 3.0 mmol) were added to anhydrous acetonitrile (10.0 mL), and the mixture was heated to 60°C for 30 min. The reaction was monitored by TLC. The organic phase was concentrated under vacuum, and the residue was further purified by silica gel column chromatography using dichloromethane and methanol as the mobile phase. Flash column chromatography was then used to purify the residue, yielding a yellow product (2.22 g, 83% yield). Sodium borohydride (0.11 g, 3.0 mmol) and potassium carbonate (0.41 g, 3.0 mmol) were added to anhydrous acetonitrile (10.0 mL) of the yellow product (0.36 g, 1.0 mmol), and the mixture was heated to 71 °C to react 30 molecules. Crotonyl bromide (second starting material, 0.40 g, 3.0 mmol) was added to the reaction system. The reaction progress was monitored by TLC. The mixture was cooled until the solid completely precipitated, filtered, and the filtrate was mixed with silica gel. The yellow solid was purified by Flash column chromatography using dichloromethane and methanol as the mobile phase. The NMR results of the final product are as follows:

[0062] 1 H NMR (500 MHz, DMSO-d6) δ 9.94 (s, 1H), 7.32 (d, J = 8.8 Hz, 1H), 7.28 – 7.22 (m, 2H), 7.13 (d, J = 8.8 Hz, 1H), 5.87 – 5.78 (m, 1H), 5.74 –5.56 (m, 3H), 4.69 (dt, J = 4.4, 1.1 Hz, 2H), 4.64 (ddd, J = 7.3, 4.6, 1.7Hz, 2H), 3.84 (d, J = 9.5 Hz, 6H), 3.43 (dp, J = 7.2, 1.1 Hz, 2H), 3.18(dddd, J = 14.9, 7.6, 4.6, 1.0 Hz, 2H), 1.67 (ddt, J = 5.4, 1.7, 1.0 Hz, 3H), 1.62 (dq, J = 5.4, 1.1 Hz, 3H).

[0063] 13 C NMR (125 MHz, DMSO-d6) δ 151.66, 149.44, 148.81, 147.54, 141.69,136.90, 132.82, 130.37, 129.74, 128.31, 128.26, 127.61, 126.35, 122.59,122.02, 121.05, 118.45, 112.72, 112.25, 71.64, 58.10, 56.16, 56.00, 33.10,27.95, 17.90, 17.70.

[0064] ESI + 446.57

[0065] Example 2 Synthesis of (E)-13-(2-buten-1-yl)-2,3,10-trimethoxy-9-((3-methyl-2-buten-1-yl)oxy)-5,6-dihydroisoquino[3,2-a]isoquinoline-7-onium (compound 2)

[0066]

[0067] The experimental procedure was the same as in Example 1, except that the first raw material was 3,3-dimethylallyl bromide and the second raw material was crotonyl bromide; the NMR results of the final product are as follows:

[0068] 1 H NMR (500 MHz, DMSO-d6) δ 9.94 (s, 1H), 7.32 (d, J = 8.8 Hz, 1H), 7.28 – 7.22 (m, 2H), 7.13 (d, J = 8.8 Hz, 1H), 5.88 – 5.78 (m, 1H), 5.72 –5.62 (m, 1H), 5.30 (dtq, J = 4.8, 3.2, 1.6 Hz, 1H), 4.67 – 4.61 (m, 4H), 3.84(d, J = 9.5 Hz, 6H), 3.43 (dp, J = 7.1, 1.1 Hz, 2H), 3.18 (dddd, J = 14.9,7.6, 4.6, 1.0 Hz, 2H), 1.75 (q, J = 1.1 Hz, 6H), 1.62 (dq, J = 5.4, 1.1 Hz, 3H).

[0069] 13 C NMR (125 MHz, DMSO-d6) δ 151.66, 149.44, 148.81, 147.72, 141.69,136.90, 136.42, 132.82, 130.37, 128.31, 128.26, 127.61, 122.59, 122.02,121.05, 119.36, 118.45, 112.72, 112.25, 68.79, 58.10, 56.16, 56.00, 33.10,27.95, 24.62, 19.90, 17.90.

[0070] ESI + 460.59

[0071] Example 3 Synthesis of (E)-9-(allyloxy)-13-(but-2-en-1-yl)-2,3,10-trimethoxy-5,6-dihydroisoquinolino[3,2-a]isoquinolino-7-onium (compound 3)

[0072]

[0073] The experimental procedure was the same as in Example 1, except that the first raw material was 3-bromopropene and the second raw material was crotonyl bromide; the NMR results of the final product are as follows:

[0074] 1H NMR (500 MHz, DMSO-d6) δ 9.94 (s, 1H), 7.32 (d, J = 8.8 Hz, 1H),7.28 – 7.22 (m, 2H), 7.13 (d, J = 8.8 Hz, 1H), 6.06 (tt, J = 11.2, 5.6 Hz,1H), 5.88 – 5.78 (m, 1H), 5.72 – 5.62 (m, 1H), 5.40 – 5.32 (m, 2H), 4.69 (dt,J = 5.6, 1.0 Hz, 2H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 3.84 (d, J = 9.5Hz, 6H), 3.43 (dp, J = 7.1, 1.1 Hz, 2H), 3.18 (dddd, J = 14.9, 7.6, 4.6, 1.0Hz, 2H), 1.62 (dq, J = 5.4, 1.1 Hz, 3H).

[0075] 13 C NMR (125 MHz, DMSO-d6) δ 151.66, 149.44, 148.81, 147.35, 141.69,136.90, 132.82, 132.43, 130.37, 128.31, 128.26, 127.61, 122.59, 122.02,121.05, 118.45, 117.90, 112.72, 112.25, 71.77, 58.10, 56.16, 56.00, 33.10,27.95, 17.90.

[0076] ESI + :432.54.

[0077] Example 4 Synthesis of (E)-13-(but-2-en-1-yl)-2,3,10-trimethoxy-9-(prop-2-yn-1-yloxy)-5,6-dihydroisoquino[3,2-a]isoquinoline-7-onium (compound 4)

[0078]

[0079] The experimental procedure was the same as in Example 1, except that the first raw material was 3-bromopropyne and the second raw material was crotonyl bromide; the NMR results of the final product are as follows:

[0080] 1H NMR (500 MHz, DMSO-d6) δ 9.94 (s, 1H), 7.41 – 7.36 (m, 1H), 7.28 –7.22 (m, 2H), 7.14 (d, J = 8.5 Hz, 1H), 5.87 – 5.78 (m, 1H), 5.72 – 5.62 (m,1H), 4.85 (d, J = 2.9 Hz, 2H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 3.89 (s,3H), 3.84 (d, J = 9.5 Hz, 6H), 3.43 (dp, J = 7.1, 1.1 Hz, 2H), 3.35 (d, J =6.1 Hz, 1H), 3.18 (dddd, J = 14.9, 7.6, 4.6, 1.0 Hz, 2H), 1.62 (dq, J = 5.4,1.1 Hz, 3H).

[0081] 13 C NMR (125 MHz, DMSO-d6) δ 151.66, 149.44, 148.67, 147.76, 141.74,136.90, 132.82, 130.46, 128.37, 128.26, 127.61, 122.59, 121.98, 121.01,118.52, 112.72, 112.25, 78.88, 76.69, 61.36, 58.15, 56.16, 56.00, 33.10,27.95, 17.90.

[0082] ESI + :430.52.

[0083] Example 5 Synthesis of (E)-13-(but-2-en-1-yl)-9-(hept-6-en-1-yloxy)-2,3,10-trimethoxy-5,6-dihydroisoquinolino[3,2-a]isoquinolino-7-onium (compound 5)

[0084]

[0085] The experimental procedure was the same as in Example 1, except that the first raw material was 7-bromoheptene and the second raw material was crotonyl bromide; the NMR results of the final product are as follows:

[0086] 1H NMR (500 MHz, DMSO-d6) δ 9.94 (s, 1H), 7.41 – 7.36 (m, 1H), 7.28 –7.22 (m, 2H), 7.15 (d, J = 8.8 Hz, 1H), 5.88 – 5.72 (m, 2H), 5.72 – 5.62 (m,1H), 5.10 (ddt, J = 10.3, 2.0, 1.0 Hz, 1H), 4.98 (ddt, J = 10.2, 2.1, 1.0 Hz,1H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 4.07 (t, J = 6.0 Hz, 2H), 3.89 (s,3H), 3.84 (d, J = 9.5 Hz, 6H), 3.43 (dt, J = 7.1, 1.1 Hz, 2H), 3.18 (dddd, J= 14.9, 7.6, 4.6, 1.0 Hz, 2H), 2.03 (tdt, J = 7.9, 6.8, 1.1 Hz, 2H), 1.70(qd, J = 6.8, 5.9 Hz, 2H), 1.62 (dq, J = 5.4, 1.1 Hz, 3H), 1.43 (qd, J = 6.6,5.6 Hz, 2H), 1.40 – 1.30 (m, 2H).

[0087] 13 C NMR (125 MHz, DMSO-d6) δ 151.66, 149.44, 148.88, 148.44, 141.79,138.86, 136.84, 132.82, 130.34, 128.31, 128.26, 127.61, 122.59, 122.04,121.37, 118.39, 114.60, 112.72, 112.25, 72.63, 58.10, 56.16, 56.00, 33.67,33.10, 29.05, 28.63, 27.95, 25.86, 17.90.

[0088] ESI + :488.65。

[0089] Example 6 Synthesis of (E)-13-(but-2-en-1-yl)-2,3,10-trimethoxy-9-((4-nitrobenzyl)oxy)-5,6-dihydroisoquinolino[3,2-a]isoquinolino-7-onium (compound 6)

[0090]

[0091] The experimental procedure was the same as in Example 1, except that the first raw material was p-nitrobenzyl bromide and the second raw material was crotonyl bromide; the NMR results of the final product are as follows:

[0092] 1 H NMR (500 MHz, DMSO-d6) δ 9.94 (s, 1H), 8.14 – 8.08 (m, 2H), 7.61 (dt, J = 8.1, 1.0 Hz, 2H), 7.41 – 7.36 (m, 1H), 7.28 – 7.22 (m, 2H), 7.15 (d,J = 8.8 Hz, 1H), 5.88 – 5.78 (m, 1H), 5.72 – 5.62 (m, 1H), 5.16 (t, J = 1.0Hz, 2H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 3.90 (s, 3H), 3.84 (d, J = 9.5Hz, 6H), 3.43 (dt, J = 7.1, 1.1 Hz, 2H), 3.18 (dddd, J = 14.9, 7.6, 4.6, 1.0Hz, 2H), 1.62 (dq, J = 5.4, 1.1 Hz, 3H).

[0093] 13 C NMR (125 MHz, DMSO-d6) δ 151.66, 149.44, 148.94, 148.25, 147.03,141.69, 140.63, 136.90, 132.82, 130.45, 128.72, 128.31, 128.26, 127.61,123.65, 122.59, 121.98, 121.17, 118.52, 112.72, 112.25, 73.04, 58.10, 56.16,56.00, 33.10, 27.95, 17.90.

[0094] ESI + :527.60.

[0095] Example 7 Synthesis of (E)-13-(but-2-en-1-yl)-2,3,10-trimethoxy-9-((4-methylbenzyl)oxy)-5,6-dihydroisoquinolino[3,2-a]isoquinolino-7-onium (compound 7)

[0096]

[0097] The experimental procedure was the same as in Example 1, except that the first raw material was 4-methylbenzyl bromide and the second raw material was crotonyl bromide; the NMR results of the final product are as follows:

[0098] 1 H NMR (500 MHz, DMSO-d6) δ 9.94 (s, 1H), 7.41 – 7.36 (m, 1H), 7.31 (t, J = 1.1 Hz, 1H), 7.30 – 7.22 (m, 3H), 7.15 (dd, J = 8.4, 2.6 Hz, 3H), 5.88 – 5.78 (m, 1H), 5.72 – 5.62 (m, 1H), 5.16 (t, J = 1.0 Hz, 2H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 3.84 (d, J = 9.5 Hz, 6H), 3.43 (dp, J = 7.1,1.1 Hz, 2H), 3.18 (dddd, J = 14.9, 7.6, 4.6, 1.0 Hz, 2H), 2.34 (d, J = 1.0Hz, 3H), 1.62 (dq, J = 5.4, 1.1 Hz, 3H).

[0099] 13 C NMR (125 MHz, DMSO-d6) δ 151.66, 149.44, 148.94, 147.03, 141.69,138.34, 136.90, 133.72, 132.82, 130.45, 129.09, 128.31, 128.26, 128.12,127.61, 122.59, 121.98, 121.17, 118.52, 112.72, 112.25, 73.04, 58.10, 56.16,56.00, 33.10, 27.95, 21.05, 17.90.

[0100] ESI + :496.63.

[0101] Example 8 Synthesis of (E)-13-(but-2-en-1-yl)-2,3,10-trimethoxy-9-((3-methylbenzyl)oxy)-5,6-dihydroisoquinolino[3,2-a]isoquinolino-7-onium (compound 8)

[0102]

[0103] The experimental procedure was the same as in Example 1, except that the first raw material was 3-methylbenzyl bromide and the second raw material was crotonyl bromide; the NMR results of the final product are as follows:

[0104] 1 H NMR (500 MHz, DMSO-d6) δ 9.94 (s, 1H), 7.41 – 7.36 (m, 1H), 7.34(ddq, J = 7.6, 2.2, 1.1 Hz, 1H), 7.25 (d, J = 8.2 Hz, 1H), 7.24 – 7.18 (m,3H), 7.18 – 7.08 (m, 2H), 5.87 – 5.78 (m, 1H), 5.72 – 5.62 (m, 1H), 5.14 (t,J = 1.1 Hz, 2H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 3.90 (s, 3H), 3.84 (d,J = 9.5 Hz, 6H), 3.43 (dt, J = 7.1, 1.0 Hz, 2H), 3.18 (dddd, J = 14.9, 7.6,4.6, 1.0 Hz, 2H), 2.33 (s, 3H), 1.62 (dq, J = 5.4, 1.1 Hz, 3H).

[0105] 13 C NMR (125 MHz, DMSO-d6) δ 151.66, 149.44, 148.94, 147.03, 141.69,137.42, 136.90, 135.58, 132.82, 130.45, 128.95, 128.88, 128.51, 128.31,128.26, 127.61, 126.04, 122.59, 121.98, 121.17, 118.52, 112.72, 112.25,73.92, 58.10, 56.16, 56.00, 33.10, 27.95, 21.05, 17.90.

[0106] ESI+ :596.63.

[0107] Example 9 Synthesis of (E)-13-(but-2-en-1-yl)-9-((4-isopropylbenzyl)oxy)-2,3,10-trimethoxy-5,6-dihydroisoquinolino[3,2-a]isoquinolino-7-onium (compound 9)

[0108]

[0109] The experimental procedure was the same as in Example 1, except that the first raw material was p-isopropylbenzyl bromide and the second raw material was crotonyl bromide; the NMR results of the final product are as follows:

[0110] 1 H NMR (500 MHz, DMSO-d6) δ 9.94 (s, 1H), 7.41 – 7.36 (m, 1H), 7.30 (dt, J = 7.5, 1.0 Hz, 2H), 7.28 – 7.22 (m, 2H), 7.14 (dd, J = 11.2, 8.1 Hz,3H), 5.87 – 5.78 (m, 1H), 5.72 – 5.62 (m, 1H), 5.16 (t, J = 1.0 Hz, 2H), 4.64(ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 3.90 (s, 3H), 3.84 (d, J = 9.5 Hz, 6H), 3.43(dt, J = 7.1, 1.1 Hz, 2H), 3.18 (dddd, J = 14.9, 7.6, 4.6, 1.0 Hz, 2H), 2.88(p, J = 6.7 Hz, 1H), 1.62 (dq, J = 5.4, 1.1 Hz, 3H), 1.27 (d, J = 6.6 Hz, 6H).

[0111] 13C NMR (125 MHz, DMSO-d6) δ 151.66, 149.44, 148.94, 147.03, 146.92,141.69, 136.90, 135.42, 132.82, 130.45, 128.31, 128.25, 127.61, 125.74,122.59, 121.98, 121.17, 118.52, 112.72, 112.25, 73.30, 58.10, 56.16, 56.00,33.66, 33.10, 27.95, 23.93, 17.90.

[0112] ESI + 524.68.

[0113] Example 10 Synthesis of (E)-9-([1,1'-biphenyl]-4-ylmethoxy)-13-(but-2-en-1-yl)-2,3,10-trimethoxy-5,6-dihydroisoquinolino[3,2-a]isoquinolino-7-onium (compound 10)

[0114]

[0115] The experimental procedure was the same as in Example 1, except that the first raw material was 4-bromomethylbiphenyl and the second raw material was crotonyl bromide; the NMR results of the final product are as follows:

[0116] 1H NMR (500 MHz, DMSO-d6) δ 9.94 (s, 1H), 7.68 – 7.63 (m, 2H), 7.59 (dt, J = 8.2, 1.0 Hz, 2H), 7.57 – 7.51 (m, 2H), 7.49 – 7.45 (m, 1H), 7.45 – 7.35 (m, 3H), 7.28 – 7.22 (m, 2H), 7.15 (d, J = 8.8 Hz, 1H), 5.88 – 5.78 (m,1H), 5.72 – 5.62 (m, 1H), 5.15 (t, J = 1.0 Hz, 2H), 4.64 (ddd, J = 7.3, 4.6,1.7 Hz, 2H), 3.90 (s, 3H), 3.84 (d, J = 9.5 Hz, 6H), 3.43 (dt, J = 7.1, 1.1Hz, 2H), 3.18 (dddd, J = 14.9, 7.6, 4.6, 1.0 Hz, 2H), 1.62 (dq, J = 5.4, 1.1Hz, 3H).

[0117] 13 C NMR (125 MHz, DMSO-d6) δ 151.66, 149.44, 148.94, 147.03, 141.69,141.54, 139.16, 136.90, 136.21, 132.82, 130.45, 128.80, 128.78, 128.66,128.31, 128.26, 127.68, 127.61, 127.21, 122.59, 121.98, 121.17, 118.52,112.72, 112.25, 73.50, 58.11, 56.16, 56.00, 33.10, 27.95, 17.90.

[0118] ESI + :558.70.

[0119] Example 11 Synthesis of (E)-13-(but-2-en-1-yl)-9-(hept-6-yn-1-yloxy)-2,3,10-trimethoxy-5,6-dihydroisoquinolino[3,2-a]isoquinolino-7-onium (compound 11)

[0120]

[0121] The experimental procedure was the same as in Example 1, except that the first raw material was 7-bromohept-1-yne and the second raw material was crotonyl bromide; the NMR results of the final product are as follows:

[0122] 1 H NMR (500 MHz, DMSO-d6) δ 9.94 (s, 1H), 7.41 – 7.36 (m, 1H), 7.28 –7.22 (m, 2H), 7.15 (d, J = 8.8 Hz, 1H), 5.87 – 5.78 (m, 1H), 5.72 – 5.62 (m,1H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 4.06 (t, J = 6.1 Hz, 2H), 3.89 (s,3H), 3.84 (d, J = 9.5 Hz, 6H), 3.43 (dq, J = 7.1, 1.0 Hz, 2H), 3.18 (dddd, J= 14.9, 7.6, 4.6, 1.0 Hz, 2H), 2.11 (td, J = 5.8, 3.0 Hz, 2H), 2.06 (t, J =2.9 Hz, 1H), 1.75 (p, J = 6.3 Hz, 2H), 1.62 (dq, J = 5.4, 1.1 Hz, 3H), 1.57 –1.46 (m, 4H).

[0123] 13 C NMR (125 MHz, DMSO-d6) δ 151.66, 149.44, 148.88, 148.44, 141.79,136.84, 132.82, 130.34, 128.31, 128.26, 127.61, 122.59, 122.04, 121.37,118.39, 112.72, 112.25, 83.79, 72.63, 69.15, 58.11, 56.16, 56.00, 33.10,29.03, 27.95, 27.85, 25.70, 17.92, 17.90.

[0124] ESI + :486.63.

[0125] Example 12 Synthesis of (E)-13-(but-2-en-1-yl)-9-((5-cyanopentyl)oxy)-2,3,10-trimethoxy-5,6-dihydroisoquinolino[3,2-a]isoquinolino-7-onium (compound 12)

[0126]

[0127] The experimental procedure was the same as in Example 1, except that the first raw material was 7-bromoheptanenitrile and the second raw material was crotonyl bromide; the NMR results of the final product are as follows:

[0128] 1 H NMR (500 MHz, DMSO-d6) δ 9.94 (s, 1H), 7.41 – 7.36 (m, 1H), 7.28 –7.22 (m, 2H), 7.15 (d, J = 8.8 Hz, 1H), 5.88 – 5.78 (m, 1H), 5.72 – 5.62 (m,1H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 4.06 (t, J = 6.1 Hz, 2H), 3.89 (s,3H), 3.84 (d, J = 9.5 Hz, 6H), 3.43 (dt, J = 7.1, 1.1 Hz, 2H), 3.18 (dddd, J= 14.9, 7.6, 4.6, 1.0 Hz, 2H), 2.39 (t, J = 5.9 Hz, 2H), 1.75 (p, J = 6.2 Hz, 2H), 1.67 (p, J = 5.9 Hz, 2H), 1.63 – 1.59 (m, 3H), 1.49 (p, J = 6.0 Hz, 2H).

[0129] 13 C NMR (125 MHz, DMSO-d6) δ 151.66, 149.44, 148.88, 148.44, 141.79,136.84, 132.82, 130.34, 128.31, 128.26, 127.61, 122.59, 122.04, 121.37,119.41, 118.39, 112.72, 112.25, 72.63, 58.10, 56.16, 56.00, 33.10, 29.02,27.95, 25.52, 24.65, 17.90, 17.06.

[0130] ESI+ :487.62.

[0131] Example 13: 9-((3,5-dimethoxybenzyl)oxy)-2,3,10-trimethoxy-13-(3-methyl-2-butenyl)-5,6-dihydroisoquinolino[3,2-a]isoquinolino-7-ammonium (Compound 13)

[0132]

[0133] The experimental procedure was the same as in Example 1, except that the first raw material was 1-bromo-3-methyl-2-butene and the second raw material was 3,5-dimethoxybenzyl bromide. The NMR results of the final product are as follows:

[0134] 1 H NMR (400 MHz, DMSO) δ 9.89 (s, 1H), 8.26 (d, J = 9.4 Hz, 1H), 7.96 (d, J = 9.4 Hz, 1H), 7.29 (s, 1H), 7.20 (s, 1H), 6.77 (d, J = 2.3 Hz, 2H), 6.47 (q, J = 2.8 Hz, 1H), 5.42 (t, J = 5.1 Hz, 1H), 5.31 (s, 2H), 4.88 – 4.80(m, 2H), 4.11 (s, 3H), 3.92 (d, J = 23.6 Hz, 5H), 3.74 (d, J = 5.5 Hz, 11H),3.15 (dt, J = 11.7, 6.4 Hz, 2H), 1.79 (s, 5H), 1.40 (s, 1H), 1.32 – 1.21 (m,3H).

[0135] 13 C NMR (101 MHz, DMSO) δ 160.91, 160.78, 151.22, 150.90, 147.78,145.17, 142.98, 139.05, 136.78, 136.12, 133.00, 132.74, 132.24, 126.53,122.36, 121.96, 121.88, 119.36, 113.66, 111.66, 107.07, 104.43, 100.66,98.94, 75.78, 63.27, 57.48, 56.35, 56.07, 55.77, 55.51, 29.91, 27.23, 25.60, 18.88.

[0136] ESI + : 556.68.

[0137] Example 14: Synthesis of 2,3,10-trimethoxy-9-((4-isopropylbenzyl)oxy)-13-(3-methyl-2-butenyl)-5,6-dihydroisoquinolino[3,2-a]isoquinoline-7-ammonium (compound 14)

[0138]

[0139] The experimental procedure was the same as in Example 1, except that the first raw material was 1-bromo-3-methyl-2-butene and the second raw material was p-isopropylbenzyl bromide. The NMR results of the final product are as follows:

[0140] 1 H NMR (400 MHz, DMSO) δ 9.78 (s, 1H), 8.28 (d, J = 9.4 Hz, 1H), 7.99 (d, J = 9.4 Hz, 1H), 7.53 (d, J = 7.8 Hz, 2H), 7.29 (d, J = 7.4 Hz, 3H), 7.22(s, 1H), 5.44 (d, J = 4.9 Hz, 1H), 5.34 (s, 2H), 4.84 (t, J = 5.8 Hz, 2H), 4.13 (s, 3H), 3.97 (s, 2H), 3.92 (s, 4H), 3.75 (s, 3H), 3.15 (t, J = 5.8 Hz, 2H), 2.91 (p, J = 6.9 Hz, 1H), 1.81 (d, J = 3.1 Hz, 6H), 1.34 – 1.17 (m,10H).

[0141] 13 C NMR (101 MHz, DMSO) δ 151.23, 151.03, 149.27, 147.79, 145.12,143.15, 136.71, 136.12, 134.29, 133.01, 132.74, 132.22, 129.55, 126.71,126.53, 122.36, 121.98, 121.87, 119.34, 113.63, 111.65, 75.91, 57.57, 57.43,56.32, 56.07, 33.68, 29.89, 27.23, 25.60, 24.31, 18.87.

[0142] ESI + 538.71

[0143] Example 15: Synthesis of 2,3,10-trimethoxy-9-((4-methylbenzyl)oxy)-13-(3-methyl-2-enylbutyl)-5,6-dihydroisoquinolino[3,2-a]isoquinoline-7-ammonium (compound 15)

[0144]

[0145] The experimental procedure was the same as in Example 1, except that the first raw material was 1-bromo-3-methyl-2-butene and the second raw material was 4-methylbenzyl bromide. The NMR results of the final product are as follows:

[0146] 1 H NMR (400 MHz, DMSO) δ 9.77 (s, 1H), 8.25 (d, J = 9.4 Hz, 1H), 7.95 (d, J = 9.4 Hz, 1H), 7.52 – 7.45 (m, 2H), 7.29 (s, 1H), 7.25 – 7.16 (m, 3H),5.46 – 5.39 (m, 1H), 5.34 (s, 2H), 4.82 (t, J = 5.9 Hz, 2H), 4.11 (s, 3H),3.98 – 3.92 (m, 2H), 3.90 (s, 3H), 3.73 (s, 3H), 3.14 (s, 2H), 2.30 (s, 3H), 1.79 (t, J = 1.9 Hz, 6H), 1.23 (s, 1H).

[0147] 13 C NMR (101 MHz, DMSO) δ 158.81, 158.46, 158.11, 151.23, 150.92,147.78, 145.08, 143.06, 138.29, 136.76, 136.10, 133.83, 133.02, 132.76,132.24, 129.48, 129.41, 126.51, 122.34, 121.95, 121.77, 119.35, 113.66,111.63, 75.82, 57.60, 57.41, 56.31, 56.05, 29.90, 27.24, 25.57, 21.28, 18.85.

[0148] ESI+ 510.26

[0149] Example 16: Synthesis of 9-([1,1'-biphenyl]-4-ylmethoxy)-2,3,10-trimethoxy-13-(3-methyl-2-butenyl)-5,6-dihydroisoquino[3,2-a]isoquinoline-7-ammonium (compound 16)

[0150]

[0151] The experimental procedure was the same as in Example 1, except that the first raw material was 1-bromo-3-methyl-2-butene and the second raw material was 4-bromomethylbiphenyl. The NMR results of the final product are as follows:

[0152] 1 H NMR (400 MHz, DMSO) δ 9.84 (s, 1H), 8.27 (d, J = 9.4 Hz, 1H), 7.98 (d, J = 9.4 Hz, 1H), 7.76 – 7.64 (m, 5H), 7.51 – 7.42 (m, 2H), 7.45 – 7.33(m, 1H), 7.29 (s, 1H), 7.19 (s, 1H), 5.42 (s, 3H), 4.88 – 4.81 (m, 2H), 4.13(s, 3H), 3.95 (d, J = 5.1 Hz, 2H), 3.89 (s, 3H), 3.73 (s, 3H), 3.17 – 3.08(m, 1H), 2.71 (d, J = 17.1 Hz, 1H), 1.79 (s, 5H), 1.23 (s, 1H).

[0153] 13C NMR (101 MHz, DMSO) δ 158.80, 158.45, 151.23, 150.96, 147.79,145.10, 143.07, 140.69, 140.08, 136.80, 136.12, 136.06, 133.05, 132.79,132.23, 129.96, 129.46, 128.12, 127.14, 127.12, 126.54, 122.35, 121.95,121.91, 119.34, 113.66, 111.63, 75.64, 57.59, 57.46, 56.31, 56.05, 38.71, 36.24, 29.90, 27.23, 25.57, 18.86, 9.05.

[0154] ESI + 572.28

[0155] Example 17: Synthesis of 9-((3,5-dimethylbenzyl)oxy)-2,3,10-trimethoxy-13-(3-methyl-2-butenyl)-5,6-dihydroisoquinoline[3,2-a]isoquinoline-7-ammonium (compound 17)

[0156]

[0157] The experimental procedure was the same as in Example 1, except that the first raw material was 1-bromo-3-methyl-2-butene and the second raw material was 3,5-dimethylbenzyl bromide. The NMR results of the final product are as follows:

[0158] 1 H NMR (400 MHz, DMSO) δ 9.81 (s, 1H), 8.29 (d, J = 9.4 Hz, 1H), 8.00 (d, J = 9.4 Hz, 1H), 7.32 (s, 1H), 7.26 – 7.20 (m, 3H), 7.02 (s, 1H), 5.49 –5.41 (m, 1H), 5.29 (s, 2H), 4.90 – 4.82 (m, 2H), 4.14 (s, 3H), 4.01 – 3.95(m, 2H), 3.92 (s, 3H), 3.76 (s, 3H), 3.16 (t, J = 5.8 Hz, 2H), 2.32 (s, 6H), 1.82 (d, J = 2.1 Hz, 5H), 1.25 (s, 1H).

[0159] 13 C NMR (101 MHz, DMSO) δ 158.27, 151.23, 151.06, 147.79, 145.09,143.22, 137.91, 136.77, 136.72, 136.13, 133.01, 132.79, 132.27, 130.25,127.01, 126.53, 122.36, 121.98, 121.94, 119.36, 113.66, 111.65, 76.11, 57.59,57.44, 56.32, 56.07, 29.91, 27.25, 25.60, 21.33, 18.87.

[0160] ESI + 524.68

[0161] Example 18: Synthesis of 2,3,10-trimethoxy-13-(3-methyl-2-butenyl)-9-((3-methyl-2-enylbutyl)oxy)-5,6-dihydroisoquinoline[3,2-a]isoquinoline-7-ammonium (compound 18)

[0162]

[0163] The experimental procedure was the same as in Example 1, except that the first raw material was 1-bromo-3-methyl-2-butene and the second raw material was 1-bromo-3-methyl-2-butene. The NMR results of the final product are as follows:

[0164] 1 H NMR (400 MHz, DMSO) δ 9.86 (s, 1H), 8.25 (d, J = 9.4 Hz, 1H), 7.97(d, J = 9.4 Hz, 1H), 7.32 (s, 1H), 7.22 (s, 1H), 5.71 – 5.62 (m, 1H), 5.46(d, J = 4.9 Hz, 1H), 4.89 (d, J = 7.1 Hz, 4H), 4.10 (s, 3H), 3.98 (s, 2H), 3.92 (s, 3H), 3.76 (s, 3H), 3.17 (t, J = 5.9 Hz, 2H), 1.82 (s, 5H), 1.73 (d,J = 14.1 Hz, 6H), 1.34 – 1.25 (m, 1H), 1.25 (s, 1H), 1.19 (s, 1H).

[0165] 13C NMR (101 MHz, DMSO) δ 151.20, 150.93, 147.76, 145.19, 143.48,139.40, 136.77, 136.10, 133.03, 132.74, 132.27, 126.40, 122.38, 122.12,121.52, 120.34, 119.41, 113.67, 111.62, 70.79, 57.51, 57.38, 56.32, 56.06,29.91, 27.25, 25.96, 25.59, 18.87, 18.38.

[0166] ESI + 474.62

[0167] Example 19: Synthesis of 9-(allyloxy)-2,3,10-trimethoxy-13-(3-methyl-2-butenyl)-5,6-dihydroisoquino[3,2-a]isoquinoline-7-ammonium (compound 19)

[0168]

[0169] The experimental procedure was the same as in Example 1, except that the first raw material was 1-bromo-3-methyl-2-butene and the second raw material was 3-chloropropene. The NMR results of the final product are as follows:

[0170] 1 H NMR (400 MHz, DMSO) δ 9.90 (s, 1H), 8.25 (d, J = 9.4 Hz, 1H), 7.96 (d, J = 9.4 Hz, 1H), 7.30 (s, 1H), 7.21 (s, 1H), 6.25 (ddt, J = 16.7, 10.2,6.1 Hz, 1H), 5.49 – 5.39 (m, 2H), 5.29 (dd, J = 10.3, 1.7 Hz, 1H), 4.92 –4.86 (m, 4H), 4.08 (s, 3H), 3.99 – 3.93 (m, 2H), 3.90 (s, 3H), 3.74 (s, 3H), 3.16 (t, J = 5.8 Hz, 2H), 3.05 (qd, J = 7.3, 4.8 Hz, 1H), 1.80 (d, J = 1.6Hz, 6H), 1.21 (t, J = 7.3 Hz, 2H).

[0171] 13C NMR (101 MHz, DMSO) δ 151.21, 150.76, 147.76, 145.17, 143.10,136.85, 136.10, 134.21, 133.05, 132.74, 132.31, 126.54, 122.40, 121.93,121.69, 119.81, 119.39, 113.68, 111.66, 75.13, 57.52, 57.45, 56.35, 56.08,45.72, 29.92, 27.22, 25.60, 18.90, 8.87.

[0172] ESI + 446.57

[0173] Example 20: Synthesis of 2,3,10-trimethoxy-13-(3-methyl-2-butenyl)-9-(prop-2-yn-1-oxy)-5,6-dihydroisoquinolino[3,2-a]isoquinoline-7-ammonium (compound 20)

[0174]

[0175] The experimental procedure was the same as in Example 1, except that the first raw material was 1-bromo-3-methyl-2-butene and the second raw material was 3-chloropropyne. The NMR results of the final product are as follows:

[0176] 1 H NMR (400 MHz, DMSO) δ 10.10 (s, 1H), 8.42 (d, J = 9.4 Hz, 1H), 8.14 (d, J = 9.4 Hz, 1H), 7.44 (s, 1H), 7.35 (s, 1H), 5.59 (t, J = 4.8 Hz,1H), 5.25 (d, J = 2.4 Hz, 2H), 5.03 (t, J = 5.7 Hz, 2H), 4.23 (s, 3H), 4.14 –4.08 (m, 2H), 4.03 (s, 3H), 3.87 (s, 3H), 3.80 (t, J = 2.4 Hz, 1H), 3.29 (dd,J = 11.3, 5.3 Hz, 3H), 1.93 (d, J = 1.8 Hz, 6H), 1.38 (s, 1H), 1.36 (s, 0H).

[0177] 13C NMR (101 MHz, DMSO) δ 151.25, 151.00, 147.77, 145.14, 141.75,136.99, 136.14, 133.01, 132.86, 132.30, 126.50, 122.44, 122.37, 122.21,119.34, 113.70, 111.66, 80.45, 79.22, 61.55, 57.56, 57.52, 56.35, 56.08,29.95, 27.20, 25.59, 18.89.

[0178] ESI + 444.55

[0179] Example 21: Synthesis of 2,3,10-trimethoxy-13-(3-methyl-2-butenyl)-9-(naphth-1-ylmethoxy)-5,6-dihydroisoquino[3,2-a]isoquinoline-7-ammonium (compound 21)

[0180]

[0181] The experimental procedure was the same as in Example 1, except that the first raw material was 1-bromo-3-methyl-2-butene and the second raw material was 1-bromomethylnaphthalene. The NMR results of the final product are as follows:

[0182] 1 H NMR (400 MHz, DMSO) δ 9.61 (s, 1H), 8.49 (d, J = 8.4 Hz, 1H), 8.29 (d, J = 9.5 Hz, 1H), 8.06 – 7.92 (m, 3H), 7.78 – 7.66 (m, 2H), 7.64 – 7.49(m, 2H), 7.27 (s, 1H), 7.18 (s, 1H), 5.85 (s, 2H), 5.41 (d, J = 5.0 Hz, 1H), 4.66 (t, J = 5.8 Hz, 2H), 4.15 (s, 3H), 3.94 (s, 2H), 3.89 (s, 3H), 3.73 (s,3H), 3.52 (s, 1H), 3.08 (t, J = 5.8 Hz, 2H), 1.84 – 1.76 (m, 6H), 1.23 (s,2H).

[0183] 13C NMR (101 MHz, DMSO) δ 151.23, 151.18, 147.79, 144.91, 143.05,136.68, 136.12, 133.73, 133.08, 132.82, 132.58, 132.13, 131.86, 129.80,129.04, 128.63, 127.20, 126.60, 126.50, 125.85, 124.68, 122.32, 122.05,121.85, 119.27, 113.62, 111.64, 73.99, 57.58, 57.45, 56.32, 56.05, 29.90, 27.20, 25.59, 18.86.

[0184] ESI + : 546.69.

[0185] Example 22: Synthesis of 9-(hept-6-en-1-oxy)-2,3,10-trimethoxy-13-(3-methyl-2-butenyl)-5,6-dihydroisoquinolino[3,2-a]isoquinoline-7-ammonium (compound 22)

[0186]

[0187] The experimental procedure was the same as in Example 1, except that the first raw material was 1-bromo-3-methyl-2-butene and the second raw material was 7-bromo-1-heptene. The NMR results of the final product are as follows:

[0188] 1H NMR (400 MHz, DMSO) δ 9.81 (s, 1H), 8.23 ​​(d, J = 9.4 Hz, 1H), 7.95 (d, J = 9.4 Hz, 1H), 7.30 (s, 1H), 7.21 (s, 1H), 5.84 (ddt, J = 17.0, 10.3, 6.7 Hz, 1H), 5.43 (t, J = 5.0 Hz, 1H), 5.04 (dq, J = 17.2, 1.8 Hz, 1H), 4.97 (ddt, J = 10.2, 2.3, 1.2 Hz, 1H), 4.87 (t, J = 5.8 Hz, 2H), 4.30 (t, J = 6.8Hz, 2H), 4.06 (s, 3H), 3.99 – 3.93 (m, 2H), 3.82 (d, J = 64.0 Hz, 6H), 3.16(t, J = 5.9 Hz, 2H), 2.14 – 2.04 (m, 2H), 1.90 (t, J = 7.2 Hz, 2H), 1.80 (d,J = 1.7 Hz, 5H), 1.50 (tq, J = 7.5, 4.0 Hz, 4H), 1.25 (s, 1H), 1.23 (s, 2H).

[0189] 13 C NMR (101 MHz, DMSO) δ 151.21, 150.69, 147.77, 145.05, 143.92,139.20, 136.82, 136.10, 133.12, 132.76, 132.29, 126.63, 122.40, 121.76,121.46, 119.41, 115.30, 113.68, 111.66, 74.79, 57.59, 57.45, 56.36, 56.09,33.64, 29.92, 29.77, 29.47, 28.51, 27.22, 25.61, 25.28, 18.90, 14.43.

[0190] ESI + : 502.67.

[0191] Example 23: Synthesis of 2,3,10-trimethoxy-9-((2-methylpropenyl)oxy)-13-(3-methyl-2-butenyl)-5,6-dihydroisoquinoline[3,2-a]isoquinoline-7-ammonium (compound 23)

[0192]

[0193] The experimental procedure was the same as in Example 1, except that the first raw material was 1-bromo-3-methyl-2-butene and the second raw material was 3-bromo-2-methylpropene. The NMR results of the final product are as follows:

[0194] 1 H NMR (400 MHz, DMSO) δ 9.79 (s, 1H), 8.25 (d, J = 9.4 Hz, 1H), 7.96 (d, J = 9.4 Hz, 1H), 7.30 (s, 1H), 7.21 (s, 1H), 5.47 – 5.41 (m, 1H), 5.22 –5.17 (m, 1H), 5.05 (t, J = 1.9 Hz, 1H), 4.91 – 4.84 (m, 2H), 4.79 (s, 2H), 4.07 (s, 3H), 3.97 (s, 2H), 3.89 (s, 3H), 3.74 (s, 3H), 3.15 (t, J = 5.9 Hz, 2H), 1.91 (s, 3H), 1.80 (d, J = 1.6 Hz, 6H), 1.32 – 1.20 (m, 2H).

[0195] 13 C NMR (101 MHz, DMSO) δ 151.23, 150.57, 147.77, 144.88, 143.34,141.39, 136.89, 136.11, 133.18, 132.83, 132.31, 126.71, 122.40, 121.62,119.38, 114.45, 113.68, 111.66, 77.84, 57.67, 57.46, 56.34, 56.08, 29.94,27.23, 25.60, 20.17, 18.89.

[0196] ESI + : 460.59.

[0197] Example 24

[0198] 1. In vitro activity assay method:

[0199] The MIC of the target compound against Helicobacter pylori was determined using the agar dilution method according to the Clinical and Laboratory Standards Institute (CLSI) guideline (M45). Antimicrobial susceptibility testing was performed using the agar dilution method and a multi-point inoculation system, following CLSI standards. A stock solution of 3840 μg / mL was diluted twice with CAMH broth to 1920, 960, 480, 240…0.94, 0.47 μg / mL. Then, 1 mL of each concentration was taken and placed in a petri dish, and 14 mL of melted MH blood agar medium (containing 5% defibrinated sheep blood) was added. The mixture was thoroughly mixed to achieve final drug concentrations of 128, 64, 32…0.06, 0.03 μg / mL. After inoculating the test bacteria on Columbia blood agar (containing 5% defibrinated sheep blood) medium for 72 h, bacterial colonies were scraped and thoroughly suspended in physiological saline. Before inoculation, the bacterial culture was adjusted to 2 McFarland turbidity (approximately 2 × 10⁻⁶) using a turbidimeter. 7 The bacterial suspension (CFU / mL) was inoculated onto a series of plates containing different concentrations of the drug using a multi-point inoculation device. The inoculation amount at each site was approximately 2 × 10⁻⁶ CFU / mL. 4 CFU is determined by placing the plate in a sealed incubator with a microaerophilic pack and incubating at 35 °C for 72 h. The minimum concentration of the drug in the sterile plate is the MIC.

[0200] Table 1 lists the in vitro activity (µg / mL) of the derivatives against four clinically collected clarithromycin / metronidazole-resistant strains.

[0201]

[0202] Example 25

[0203] Antibacterial activity of spherical drug-resistant bacteria

[0204] 1. Methods for inducing cocci

[0205] Helicobacter pylori strain SS1 was cultured overnight to mid-logarithmic growth using Brucella broth (containing 10% FBS). The bacterial culture was concentrated to an OD600 of 5.0 for later use. Amoxicillin-induced coccidial Helicobacter pylori system was prepared according to Table 1, with a negative control group included. The cultures were incubated at 37°C in a tri-gas incubator with shaking for 24 h. After 24 h of induction, slides were prepared and Gram-stained to observe bacterial morphology. Experiments could only continue when 100% of the bacteria in the amoxicillin-treated group were coccidial.

[0206] 2. Method for determining the activity of compound-forming cocci.

[0207] Naturally induced Helicobacter pylori SS1 spherical strains were Gram-stained and examined under a microscope to confirm that all SS1 strains had transformed into spherical bacteria, which remained spherical after 12 hours of incubation. The bacteria were resuspended in Brucella medium containing 2% FBS by centrifugation to an OD600 of 5. Two mL of the test compound (0.03–128 µg / mL) was added to each culture tube and incubated for 24 h. 100 µL of the bacterial suspension was spread onto BHI agar plates containing 5% defibrinated sheep blood, with a normal control group included. The plates were incubated at 37 °C for 72 h under microaerobic conditions (10% CO2).

[0208] Table 2. Minimum bactericidal concentrations (MBC: μg / mL) of 9-,13-disubstituted palmatine derivatives and control drugs against Helicobacter pylori.

[0209] All compounds showed good efficacy against *Helicobacter pylori* with spherical deformities. The compound with the weakest activity was comparable in efficacy to the aforementioned 4b compound, i.e., the 3-,13-disubstituted compound. This demonstrates that all compounds in this application exhibit better bactericidal effects against spherical bacteria than 4a-4d. (See the antibacterial diagram below.) Figure 1 .

[0210] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, it is intended that all variations falling within the meaning and scope of equivalents of the claims be included within the present invention.

[0211] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A palmatine derivative, characterized in that, The general formula of the palmatine derivative is as follows: R1 is selected from isopentenyl, 2-butenyl; The R2 is selected from C3-C8 alkenyl, C3-C8 alkynyl, benzyl group substituted with one or two independently selected C1-C4 alkyl, C1-C4 alkoxy or phenyl or nitro groups, and naphthylmethyl.

2. The palmatine derivative according to claim 1, characterized in that, R2 is selected from the following substituents: ， ， ， ， ， ， ， ， ， ， ， ， ， ， , and .

3. The palmatine derivative according to claim 1, characterized in that, The palmatine derivative is selected from the following compounds: ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， and .

4. A pharmaceutical composition comprising the palmatine derivative or a pharmaceutically acceptable salt thereof as described in any one of claims 1–3, and at least one pharmaceutically acceptable excipient.

5. A method for preparing the palmatine derivative of claim 1, characterized in that, The preparation method consists of the following steps: 1) Take palmatine hydrochloride, heat to remove the methyl group at the 9-position, add acid solution to acidify, and obtain a dark red solid; 2) The dark red solid obtained in step 1) is added to an aprotic solvent, a catalyst and a 9-substituent starting material are added, and after heating to cause a 9-substitution reaction, the organic phase is concentrated and the residue is concentrated by column chromatography to obtain the nucleophilic substitution product. 3) Add an aprotic solvent, a reducing agent and a catalyst to the substitution product in step 2), heat to activate, add the 13-substituent raw material to carry out the 13-substitution reaction, after the reaction is completed, cool until the solid is completely precipitated, filter, and purify the filtrate by chromatographic column to obtain the palmatine derivative.

6. The preparation method according to claim 5, characterized in that, The order of steps for the 9-position substitution reaction and the 13-position substitution reaction is interchangeable.

7. The use of the palmatine derivative of any one of claims 1-3 in the preparation of a medicament for treating diseases related to Helicobacter pylori infection.

8. The application according to claim 7, characterized in that, The diseases related to Helicobacter pylori infection include Helicobacter pylori infection, as well as functional dyspepsia, gastritis, peptic ulcer, gastric mucosa-associated lymphoid tissue lymphoma and gastric cancer caused by Helicobacter pylori infection.

Images