Bis(benzylisoquinoline) alkaloids as TLR4 agonist

Abstract

The present invention discloses bis(benzylisoquinoline) alkaloid compounds, their synthesis, a combination and composition for activity against TLR4 receptors. The compounds of the present invention also show synergistic effect with other antibiotics.

Description

[0001] FIELD OF INVENTION

[0002] The present invention relates to the field of synthetic pharmaceutical chemistry. In particular, it relates the synthesis of functionalized Bis (benzylisoquinoline) alkaloids as TLR4 agonists.

[0003] BACKGROUND OF INVENTION

[0004] Toll-like receptors (TLRs) play a key role in initiating innate and adaptive immune responses, with TLR4 recognizing bacterial components. Unlike other TLRs, TLR3 recruits TRIF exclusively, while TLR4 activates both MyD88 and TRIF, initiating downstream signaling. MyD88 activation triggers NF-kB, leading to pro-inflammatory cytokine production, aiding infection clearance. TLR4 activation induces an anti-bacterial Thl immune response. Understanding pathogen-PRR interactions and immunity informs therapeutic development.

[0005] A bis (benzylisoquinoline) alkaloid, Fangchinoline (Fen) is known to have the ability of inhibiting virus replication. It has been previously shown that Fen exhibits efficacy against HIV, cancer, and arthritis. Combined Effects of Fangchinoline is known for certain bioactivity. However, the TLR4 activity of the compounds is not known.

[0006] The present invention examines new compounds, having Fangchinoline like structure for TLR4 activity. As, there is a need of the new compounds that can act as TLR4 agonist by inhibiting bacterial infections.

[0007] OBJECT OF THE INVENTION

[0008] An object of the invention is to provide functionalized bis(benzylisoquinoline) alkaloids as a TLR4 agonist, a process for the synthesis of functionalized bis(benzylisoquinoline) alkaloids, their combination, composition and uses thereof.

[0009] SUMMARY OF INVENTION

[0010] The present disclosure relates to TLR4 specific functionalized bis(benzylisoquinoline) alkaloid compounds along with their pharmaceutically acceptable salts and isomers of chemical Formula I;

[0011]

[0012] FORMULA I

[0013] Wherein:

[0014] A can be a lone pair, or can be selected from a group comprising

[0015]

[0016] Ri and R2 are selected from a group of hydrogen, halogen, cyano, nitro, substituted or unsubstituted haloalkyl, substituted or unsubstituted alkyl and substituted or unsubstituted cycloalkyl; or B(OH)2, NRaRb, -NRaC(O)Rb, or - NRaS(O)Rb, ORa;

[0017] Raand Rb, which may be same or different at each occurrence, are independently selected from the group consisting of hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkyl / alkyl, substituted or unsubstituted aryl, substituted or unsubstituted aryl / alkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaryl / alkyl, and substituted or unsubstituted heterocyclyl / alkyl; -C(O)ORa, -C(O)Ra, -C(O)NRa, and -S(O)Ra;

[0018] The present invention discloses methods to synthesize functionalized bis (benzylisoquinoline) alkaloids to act as a TLR4 agonist that promotes the nitric oxide (NO) production through Myd88-NF-kB signaling pathway and promote Thl response ((IL-6, IL-12, IFN-y, IL-17)).

[0019] The present invention discloses a combination of bis (benzylisoquinoline) alkaloid compounds and structurally similar compounds. The present invention further discloses a composition comprising functionalized bis (benzylisoquinoline) alkaloids or the combination along with the pharmaceutically acceptable adjuvants and excipients.

[0020] The present invention further discloses the use of the compounds, combination and composition in treatment of intra cellular bacterial infections by diminishing the bacterial burden and inflammation leading to recovery of animals from Salmonella infection.

[0021] BRIEF DESCRIPTION OF FIGURES

[0022] Figure 1 depicts Fangchinoline (Fen) shows no direct cytotoxicity on cells.

[0023] Figure 2 depicts Fen and compound 1001 inhibits Salmonella growth inside the macrophage.

[0024] Figure 3 depicts that Fangchinoline (Fen) acts as a TLR4 agonist.

[0025] Figure 4 depicts the synergistic effect of Fen and compound 1001 in enhancing antibiotic function and clearing salmonella infection.

[0026] Figure 5 depicts compound 1001 shows synergistic effect with conventional antibiotic.

[0027] Figure 6 depicts combinatorial treatment of Fen with Ciprofloxacin / Enrofloxacin reduces ulcers and inflammation in Salmonella infected colon tissues.

[0028] Figure 7 depicts TLR4 agonistic activity of the compounds of present invention.

[0029] DETAILED DESCRIPTION OF INVENTION

[0030] The present disclosure relates to TLR4 specific functionalized bis (benzylisoquinoline) alkaloid compounds along with their pharmaceutically acceptable salts and isomers of chemical Formula I;

[0031]

[0032] FORMULA I

[0033] Wherein: A can be a lone pair, or can be selected from a group comprising

[0034]

[0035] Ri and R2 are selected from a group of hydrogen, halogen, cyano, nitro, substituted or unsubstituted haloalkyl, substituted or unsubstituted alkyl and substituted or unsubstituted cycloalkyl; or B(OH)2, NRaRb, -NRaC(O)Rb, or - NRaS(O)Rb, ORa;

[0036] Raand Rb, which may be same or different at each occurrence, are independently selected from the group consisting of hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkyl / alkyl, substituted or unsubstituted aryl, substituted or unsubstituted aryl / alkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaryl / alkyl, and substituted or unsubstituted heterocyclyl / alkyl; -0(0)0113, -0(0)10, -C(0)NRa, and -8(0)10.

[0037] The present disclosure relates to TLR4 specific functionalized bis(benzylisoquinoline) alkaloid compounds along with their pharmaceutically acceptable salts and isomers of chemical Formula II;

[0038]

[0039] FORMULA II

[0040] Wherein: A can be a lone pair, or can be selected from a group comprising

[0041]

[0042] Ra, which may be same or different at each occurrence, is independently selected from the group consisting of substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted haloalkyl, substituted or unsubstituted cycloalkyl, -C(O)ORa, -C(O)Ra, -C(O)NRa, and -S(O)Ra;

[0043] R2 may be hydrogen, halogen, cyano, nitro, substituted or unsubstituted haloalkyl, substituted or unsubstituted alkyl and substituted or unsubstituted cycloalkyl; or B(OH)2, NRbRc, -NRbC(O)Rc, or - NRbS(O)Rc;

[0044] Rb and Rc, which may be same or different at each occurrence, are independently selected from the group consisting of hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkyl / alkyl, substituted or unsubstituted aryl, substituted or unsubstituted aryl / alkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaryl / alkyl, and substituted or unsubstituted

[0045]

[0046] COCH2CI ;

[0047] n is 6-15.

[0048] The compounds disclosed herein and their pharmaceutically acceptable salts can exist as neutral compounds, acidic and basic salts, single stereoisomers, racemates, and as mixtures of enantiomers and diastereomers. The compounds disclosed herein can also exist as geometric isomers. All such single stereoisomers, racemates and mixtures thereof, and geometric isomers are intended to be within the scope of the compounds disclosed herein. The present invention discloses exemplary compounds of formula I as below:

[0049] >

[0050] >

[0051]

[0052] "

[0053] >

[0054] >

[0055]

[0056] "

[0057] " >

[0058] <

[0059]

[0060]

[0061] The compounds of present invention includes:

[0062] 1) 1001: 16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34-octahydro-2,6- dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)-dibenzenacyclooctaphane-37-yl methanesulfonate;

[0063] 2) 1002: 16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34-octahydro-2,6- dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)-dibenzenacyclooctaphane-37-yl 4- methylbenzenesulfonate;

[0064] 3) 1003: 16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34-octahydro-2,6- dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)-dibenzenacyclooctaphane-37-yl acetate;

[0065] 4) 1004: 16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34-octahydro-2,6- dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)-dibenzenacyclooctaphane-37-yl nonionate;

[0066] 5) 1005: 16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34-octahydro-2,6- dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)-dibenzenacyclooctaphane-37-yl hexadecyrate;

[0067] 6) 1006: hexadecyl (16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34- octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)- dib enzenacy cl ooctaphane -37 -y 1) carb onate ;

[0068] 7) 1007: 16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34-octahydro-2,6- dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)-dibenzenacyclooctaphane-37-yl 2- (9H-fluoren-9-yl)acetate; 8) 1008: 16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34-octahydro-2,6- dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)-dibenzenacyclooctaphane-37-yl 2,4- difluorobenzoate;

[0069] 9) 1009: 37-(allyloxy)-16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34- octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)- dib enzenacy cl ooctaphane ;

[0070] 10)1010: 37-butoxy-16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34- octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)- dib enzenacy cl ooctaphane ;

[0071] 11) 1011: 37-(benzyloxy)-16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34- octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)- dib enzenacy cl ooctaphane ;

[0072] 12) 1012: 16,36,54-trimethoxy-12,32-dimethyl-37-((4-nitrobenzyl)oxy)- ll,12,13,14,31,32,33,34-octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina- 5(1, 3), 7(1 ,4)-dibenzenacyclooctaphane ;

[0073] 13) 1013: 16,36,54-trimethoxy-12,32-dimethyl-37-((2-nitrobenzyl)oxy)- ll,12,13,14,31,32,33,34-octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina- 5(1, 3), 7(1 ,4)-dibenzenacyclooctaphane ;

[0074] 14) 1014: 16,36,54-trimethoxy-37-((3-methoxybenzyl)oxy)-12,32-dimethyl- ll,12,13,14,31,32,33,34-octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina- 5(1, 3), 7(1 ,4)-dibenzenacyclooctaphane ;

[0075] In an embodiment, the invention provides method for the preparation of functionalized bis (benzylisoquinoline) alkaloid compounds that acts as TLR4 specific agonist. The method comprising the steps of;

[0076] i. dissolving fangchinoline in anhydrous CH2CI2 or ethyl acetate (EtOAc) or tetrahydrofuran (THF) or acetonitrile (MeCN) or toluene under an inert atmosphere;

[0077] ii. adding tri ethylamine and the corresponding sulfonyl chlorides or acyl chlorides to the solution of step i at a temperature of 0-5 °C;

[0078] iii. stirring the reaction mixture obtained from step ii at room temperature and quenching with ice cold water upon completion; iv. extracting the product obtained from step iii with CH2Q2, and drying of the combined organic layer over anhydrous sodium sulfate or magnesium sulfate or calcium chloride (CaCE) or potassium carbonate (K2CO3);

[0079] v. evaporating the solvent under reduced pressure to yield a crude product; vi. purification of the product of step v by silica gel column chromatography using MeOH: CH2Q2 as eluents, or preparative thin-layer chromatography to obtain pure products.

[0080] The compounds of present invention can be synthesised by general procedures as given below:

[0081]

[0082] S.No. Compound Reagent Base Solvent Yield (%)

[0083] Code

[0084] 1 1001 Et3N DCM 27

[0085]

[0086] 2 1002 Et3N DCM 64

[0087]

[0088] 3 1003 Et3N DCM 65

[0089]

[0090] 4 1004 Et3N DCM 56

[0091]

[0092] 5 1005 Et3N DCM 45

[0093]

[0094] 6 1006 Et3N DCM 87

[0095]

[0096] 7 1007 Pyridine DCM 08

[0097]

[0098] 8 1008 Et3N DCM 91

[0099]

[0100] 9 1009

[0101]

[0102] NaH DMF 63

[0103] 10 1010

[0104]

[0105] NaH DMF 14

[0106] 11 1011 NaH DMF 25

[0107]

[0108] 12 1012 NaH DMF 18

[0109]

[0110] 13 1013 NaH DMF 49

[0111]

[0112] 14 1014 NaH DMF 41

[0113]

[0114] General Procedure A : Fangchinoline (1-5 eq.) was dissolved in anhydrous CH2Q2 under an inert atmosphere. Tri ethylamine (1-5 eq.) and the corresponding sulfonyl chlorides or acyl chlorides (1-5 eq.) were then added to the solution at 0-5 °C. The reaction mixture was stirred at room temperature and quenched upon completion (monitored by TLC) with ice-cold water. The resulting product was extracted with CH2Q2, and the combined organic layer was dried over anhydrous sodium sulfate. Subsequently, the solvent was evaporated under reduced pressure, yielding a crude product that was purified by silica gel (230-400 mesh) column chromatography using MeOH: CH2Q2 as eluents, or preparative thin-layer chromatography to afford pure products.

[0115] General Procedure B: Fangchinoline (1-5 eq.) was dissolved in anhydrous CH2CI2, ethyl acetate (EtOAc), tetrahydrofuran (THF), acetonitrile (MeCN), or toluene under an inert atmosphere. Triethylamine (1-5 eq.) and the corresponding sulfonyl or acyl chloride (1-5 eq.) were then added to the solution at 0-5 °C. The reaction mixture was stirred at room temperature and quenched upon completion, as monitored by TLC, with ice-cold water. The resulting product was extracted with CH2CI2, and the combined organic layer was dried over anhydrous sodium sulfate, magnesium sulfate, calcium chloride (CaCL), or potassium carbonate (K2CO3). The solvent was evaporated under reduced pressure, yielding a crude product, which was purified by silica gel (230-400 mesh) column chromatography using MeOH: CH2Q2 as eluents, or preparative thin-layer chromatography to afford pure products. The present invention discloses combinations of compounds (1001, 1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011, 1012, 1013, 1014) and structurally similar compounds like Fen in conjunction with Fen.

[0116] In yet another embodiment, the present invention discloses combination of compound of the present invention with Fangchinoline (Fen) and Ciprofloxacin or Enrofloxacin (l-30mg / kg oral / i.p).

[0117] The present invention discloses a synergistic composition comprising the compounds of the present invention along with pharmaceutically acceptable excipients and adjuvants selected from a group comprising binders such as lactose, microcrystalline cellulose, starch, povidone, and hydroxypropyl methylcellulose (HPMC); diluents like calcium phosphate, mannitol, sorbitol, microcrystalline cellulose, and lactose; disintegrants including croscarmellose sodium, sodium starch glycolate, crospovidone, and starch; lubricants such as magnesium stearate, stearic acid, sodium stearyl fumarate, and talc; glidants like colloidal silicon dioxide and talc; preservatives such as benzalkonium chloride, benzoic acid, potassium sorbate, and sodium benzoate; stabilizers like citric acid, ascorbic acid, tocopherols, and EDTA; sweeteners including aspartame, sucralose, sorbitol, and xylitol; flavors and colors like natural and artificial flavoring agents, FD&C colorants, and caramel color; and solubilizers such as polyethylene glycol (PEG), polysorbate 80, and propylene glycol.

[0118] Additionally, the composition may include adjuvants such as aluminum salts (e.g., aluminum hydroxide, aluminum phosphate), oil-in-water emulsions like squalene-based adjuvants (e.g., MF59, AS03), cytokine modulators such as granulocyte-macrophage colony-stimulating factor (GM-CSF), Toll-Like Receptor (TLR) agonists like monophosphoryl lipid A (MPL) and CpG oligonucleotides, polysaccharides including chitosan and dextran, lipid-based adjuvants such as liposomes and phospholipids, saponins like QS-21, and polymeric adjuvants such as poly(lactic-co-glycolic acid) (PLGA) and polyinosinic-polycytidylic acid (poly-IC).

[0119] The present invention also encompasses compositions comprising the combination as disclosed in some respects along with pharmaceutically acceptable excipients and adjuvants. These compositions reflect varying doses of Fangchinoline and compound 1001 derivatives to achieve synergistic protection against Salmonella, in conjunction with a suboptimal ciprofloxacin dose, to reduce bacterial load and control infection-associated symptoms. Adjustments to concentrations can be made based on therapeutic requirements or specific infection models.

[0120] A suboptimal dose of Fangchinoline (Fen) in a range of 1-8 mg / kg along with the compounds of the present invention or their pharmaceutically acceptable salts in a range of 4-12 mg / kg (oral) and Ciprofloxacin in the range of 0.05- 0.20 mg / kg can be administered orally, intraperitonially, nasally, parenterally (intravenous, intramuscular, or subcutaneous), topically, transdermally, intravaginally, intravesically, intraci sternally, or rectally, in the form of solid, semi-solid, lyophilized powder, or liquid dosage forms, such as for example, tablets, suppositories, pills, soft elastic and hard gelatin capsules, powders, solutions, suspensions, or aerosols, or the like, preferably in unit dosage forms suitable for simple administration of precise dosages. Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. Solid dosage forms, as described above, can be prepared with coatings and shells, such as enteric coatings. Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. Compositions for rectal administrations are, for example, suppositories that can be prepared by mixing the compounds of this disclosure with, for example, suitable non-irritating excipients or carriers. They can also be administered as sterile powders for reconstitution into sterile injectable solutions or dispersions. Dosage forms for topical administration of a compound of this disclosure include ointments, powders, sprays. Ophthalmic formulations, eye ointments, powders, inhalation formulations and solutions are also contemplated for the compounds in this disclosure. Compressed gases can be used to disperse a compound of this disclosure in aerosol form.

[0121] The compounds of the present invention possess TLR4 agonist activity that is the compounds, combination of compounds and composition can be used in cancer immunotherapy, infectious disease treatment, as vaccine adjuvants, and in managing allergic, wound healing and chronic inflammatory diseases.

[0122] The present invention is illustrated by way of examples. The examples are meant for illustrative purposes only and cannot be construed to restrict the scope of the present invention, Examples of the present invention

[0123] The bis (benzylisoquinoline) alkaloid compounds of the present invention can be synthesised as follows:

[0124] SYNTHETIC EXAMPLES

[0125] Example 1: 7-O-(methanesulfonate)-fangchinoline (compound 1001): Fangchinoline (10 mg, 0.017 mmol, 1.00 eq.) was dissolved in anhydrous CH2Q2 (1 mL) under inert atmosphere. Triethylamine (3.4 pL, 0.025 mmol, 1.50 eq.) and methane sulfonylchloride (1.3 pL, 0.018 mmol, 1.10 eq.) were added to the solution at 0°C under nitrogen. The reaction mixture was stirred for 3 hours at room temperature. After the completion of the reaction (TLC based observation), ice cold water (1 mL) was added to the reaction mixture and the product formed was extracted with CH2Q2 (3x2 mL). The combined organic layer was dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and crude product obtained was purified with silica gel (230-400 mesh) column chromatography using MeOHiCHzCh as eluents to provide pure product 1 (compound 1001) in 27% yield.1H NMR (500 MHz, CDCh) 5 7.37 (dd, J= 8.2, 2.1 Hz, 1H), 7.15 (dd, J = 8.2, 2.5 Hz, 1H), 6.85 (d, J= 2.0 Hz, 2H), 6.81 (dd, J= 8.3, 2.5 Hz, 1H), 6.55 (s, 1H), 6.49 (d, J= 1.1 Hz, 1H), 6.39 (s, 1H), 6.33 (dd, J= 8.3, 2.2 Hz, 1H), 6.03 (s, 1H), 3.93 (s, 3H), 3.88 (dd, J= 11.2, 5.3 Hz, 1H), 3.79 (s, 3H), 3.71 (d, J = 9.8 Hz, 1H), 3.38 (s, 3H), 3.32 (dd, J= 12.4, 5.2 Hz, 1H), 2.94 (ddd, J= 18.0, 12.2, 6.3 Hz, 4H), 2.86 (s, 3H), 2.84 - 2.65 (m, 3H), 2.61 (s, 3H), 2.58 (d, J = 13.1 Hz, 1H), 2.47 (dd, J = 15.2, 4.1 Hz, 1H), 2.29 (s, 3H).13C NMR (125 MHz, CDCh) 8 153.76, 150.34, 149.44, 148.80, 148.31, 147.04, 142.31, 134.80, 134.47, 132.63, 132.47, 130.45, 128.31, 127.77, 123.76, 122.82, 122.21, 121.97, 121.58, 115.87, 112.60, 111.53, 106.19, 64.05, 61.40, 56.13, 56.08, 55.84, 45.07, 43.79, 42.33, 41.47, 39.16, 38.88, 30.32, 29.69, 24.47, 22.69, 22.27. MS (ESI-TOF) m / z calculated for C38H42N2O8S [M+H]+687.24, found 687.2.

[0126] Example 2: 7-O-(4-toulenesulfonate)-fangchinoline (compound 1002): Fangchinoline (10 mg, 0.017 mmol, 1.00 eq.) was dissolved in anhydrous CH2Q2 (1 mL) under inert atmosphere. Triethylamine (3.4 pL, 0.025 mmol, 1.50 eq.) and p-toulenesulfonylchloride (3.4 mg, 0.018 mmol, 1.10 eq.) were added to the solution at 0° under nitrogen. The reaction mixture was stirred for 5 hours at room temperature. After the completion of reaction (TLC based observation), ice cold water (ImL) was added to the reaction mixture and product formed was extracted with CH2Q2 (3x2 mL). The combined organic layer was dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and crude product obtained was purified with silica gel (230-400 mesh) column chromatography using MeOEECELCh to provide pure product 2 (compound 1002) in 64% yield. MS (ESI-TOF) m / z calculated for C44H46N2O8S [M+H]+763.2, found 763.2.

[0127] Example 3: 7-O-acetyl-fangchinoline (compound 1003): Fangchinoline (10 mg, 0.017 mmol, 1.00 eq.) was dissolved in anhydrous CH2CI2 (1 mL) under inert atmosphere. Triethylamine (6.8 pL, 0.049 mmol, 3.00 eq.) and acetylchloride (2.9 pL, 0.041 mmol, 2.50 eq.) were added to the solution at 0°C under nitrogen. The reaction mixture was stirred for 5 hours at room temperature. After the completion of reaction (TLC based observation), ice cold water (ImL) was added to the reaction mixture and product formed was extracted with CH2Q2 (3x2 mL). The combined organic layer was again washed with ice cold water three times and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and crude product obtained was purified with silica gel (230-400 mesh) column chromatography using MeOFLCFLCh as eluents to provide pure product 3 (compound 1003) in 65% yield. MS (ESI-TOF) m / z calculated for C39H44N2O7 [M+H]+651.7, found 651.2.

[0128] Example 4: 7-O-nonanoyl-fangchinoline (compound 1004): Fangchinoline (10 mg, 0.017 mmol, 1.00 eq.) was dissolved in anhydrous CH2CI2 (1 mL) under inert atmosphere. Triethylamine (6.8 pL, 0.049 mmol, 3.00 eq.) and nonanoyl chloride (7.2 pL, 0.041 mmol, 2.50 eq.) were added to the solution at 0°C under nitrogen. The reaction mixture was stirred for 8 hours at room temperature. After the completion of reaction (TLC based observation), ice cold water (ImL) was added to the reaction mixture and product obtained was extracted with CH2Q2 (3x2 mL). The combined organic layer was again washed with ice cold water three times and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and crude product obtained was purified with silica gel (230-400 mesh) column chromatography using MeOEECELCh as eluents to provide pure product 4 (compound 1004) in 56% yield. MS (ESI-TOF) m / z calculated for C46H56N2O7 [M+H]+749.4, found 749.3.

[0129] Example 5: 7-O-palmitoyl-fangchinoline (compound 1005): Fangchinoline (10 mg, 0.017 mmol, 1.00 eq.) was dissolved in anhydrous CH2CI2 (1 mL) under inert atmosphere. Triethylamine (6.8 pL, 0.049 mmol, 3.00 eq.) and palmitoylchloride (11.80 pL, 0.041 mmol, 2.50 eq.) were added to the solution at 0°C under nitrogen. The reaction mixture was stirred for 8 hours at room temperature. After the completion of reaction (TLC based observation), ice cold water (ImL) was added to the reaction mixture and product obtained was extracted with CH2Q2 (3x2 mL). The combined organic layer was again washed with ice cold water three times and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and crude product obtained was purified with silica gel (230-400 mesh) column chromatography using MeOEECELCh as eluents to provide pure product 5 (compound 1005) in 45% yield. MS (ESLTOF) m / z calculated for C46H56N2O7 [M / 2+H]+424.3, found 424.3.

[0130] Example 6: 7-O-(hexadecylcarbonate)-fangchinoline (compound 1006): Fangchinoline (10 mg, 0.017 mmol, 1.00 eq.) was dissolved in anhydrous CH2CI2 (1 mL) under inert atmosphere. Triethylamine (10.4 pL, 0.020 mmol, 5.00 eq.) and hexadecylchloroformate (16.1 pL, 0.048 mmol, 3.00 eq.) were added to the solution at 0°C under nitrogen. The reaction mixture was stirred for 8 hours at room temperature. After the completion of reaction (TLC based observation), ice cold water (ImL) was added to the reaction mixture and product obtained was extracted with CH2Q2 (3x2 mL). The combined organic layer was again washed with ice cold water three times and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and crude product obtained was purified with silica gel (230-400 mesh) column chromatography using MeOFLCFLCb as eluents to provide pure product 6 (compound 1006) in 87% yield. MS (ESLTOF) m / z calculated for C54H72N2O8 [M+H]+877.5, found 440.1 [M / 2+H]+.

[0131] Example 7: 7-O-(9-fluorenylmethylcarbonate)fangchinoline (compound 1007):

[0132] Fangchinoline (10 mg, 0.017 mmol, 1.00 eq.) was dissolved in anhydrous CH2CI2 (1 mL) under inert atmosphere. Pyridine (2.05 pL, 0.018 mmol, 1.10 eq.) and 9-fluorenyllmethylchloroformate (4.6 pL, 0.018 mmol, 1.10 eq.) were added to the solution at 0°C under nitrogen. The reaction mixture was stirred for 1 hour at room temperature. After the completion of reaction (TLC based observation), CuSO4solution (2mL) was added to the reaction mixture and product obtained was extracted with CH2CI2 (3x2 mL). The combined organic layer was again washed with ice cold water three times and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and crude product obtained was purified with silica gel (230-400 mesh) column chromatography using MeOEECELCh as eluents to provide pure product 7 (compound 1007) in 8% yield. HRMS (ESI-TOF-SYNAPT) m / z calculated for C52H50N2O8 [M+H]+830.3, found 831.3 .

[0133] Example 8: 7-O-(2,4-difluorobenzoyl)-fangchinoline (compound 1008): Fangchinoline (10 mg, 0.017 mmol, 1.00 eq.) was dissolved in anhydrous CH2CI2 (1 mL) under inert atmosphere. Triethylamine (6.8 pL, 0.049 mmol, 3.00 eq.) and 2,4-difhrorobenzoylchloride (4.8 pL, 0.041 mmol, 2.50 eq.) were added to the solution at 0°C under nitrogen. The reaction mixture was stirred for 5 hours at room temperature. After the completion of reaction (TLC based observation), ice cold water (ImL) was added to the reaction mixture and product obtained was extracted with CH2Q2 (3x2 mL). The combined organic layer was again washed with ice cold water three times and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and crude product obtained was purified with silica gel (230-400 mesh) column chromatography using MeOFLCELCh as eluents to provide pure product 8 (compound 1008) in 91% yield. MS (ESLTOF) m / z calculated for C44H42F2N2O7 [M+H]+749.2, found 749.4.

[0134] Example 9: 7-O-(prop-2-enyl)-fangchinoline (compound 1009): Fangchinoline (10 mg, 0.017 mmol, 1.00 eq.) was dissolved in dry DMF (0.1 mL) under inert atmosphere. Sodium hydride (0.9 mg, 0.039 mmol, 2.50 eq.) was added to solution at -10°C and the suspension was stirred. After 15 minutes, allyl chloride (0.8 pL, 0.009 mmol, 1.20 eq.) was added under nitrogen and the reaction mixture was allowed to stir for more 8 hours at room temperature. After the completion of reaction (according to TLC), the reaction mixture was quenched with ice cold water. The product obtained was extracted with CH2CI2 and the combined organic layer was dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and crude product obtained was purified by silica gel (230-400 mesh) column chromatography MeOH: CH2Q2 as eluents to provide pure product 9 (compound 1009) in 63% yield. MS (ESLTOF) m / z calculated for C40H44N2O6 [M+H]+649.3, found 649.3.

[0135] Example 10: 7-O-butyl-fangchinoline (compound 1010): Fangchinoline (10 mg, 0.017 mmol, 1.00 eq.) was dissolved in dry DMF (0.1 mL) under inert atmosphere. Sodium hydride (0.9 mg, 0.039 mmol, 2.50 eq.) was added to solution at -10°C and the suspension was stirred. After 15 minutes, iodobutane (2.5 pL, 0.009 mmol, 1.20 eq.) was added under nitrogen and the reaction mixture was allowed to stir for more 8 hours at room temperature. After the completion of reaction (according to TLC), the reaction mixture was quenched with ice cold water. The product obtained was extracted with CH2CI2 and the combined organic layer was dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and crude product obtained was purified by silica gel (230-400 mesh) column chromatography MeOH: CH2Q2 as eluents to provide pure product 10 (compound 1010) in 14% yield. MS (ESI-TOF) m / z calculated for C41H48N2O6 [M+H]+665.3, found 665.5.

[0136] Example 11: 7-O-benzyl-fangchinoline (compound 1011): Fangchinoline (10 mg, 0.017 mmol, 1.00 eq.) was dissolved in dry DMF (O.lmL) under inert atmosphere. Sodium hydride (0.9 mg, 0.039 mmol, 2.50 eq.) was added to solution at -10°C and the suspension was stirred. After 15 minutes, benzyl bromide (4.2 mg, 0.019 mmol, 1.20 eq.) was added under nitrogen and the reaction mixture was allowed to stir for more 8 hours at room temperature. After the completion of reaction (according to TLC), the reaction mixture was quenched with ice cold water. The product formed was extracted with CH2Q2 and the combined organic layer was dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and crude product obtained was purified with silica gel (230-400 mesh) chromatography using MeOH: CH2Q2 as eluents to provide pure product 11 (compound 1011) in 25% yield. MS (ESI-TOF) m / z calculated for C44H46N2O6 [M+H]+699.2, found 699.2.

[0137] Example 12: 7-O-(4-nitrobenzyl)-fangchinoline (compound 1012): Fangchinoline (10 mg, 0.017 mmol, 1.00 eq.) was dissolved in dry DMF (O.lmL) under inert atmosphere. Sodium hydride (0.9 mg, 0.039 mmol, 2.50 eq.) was added to solution at -10°C and the suspension was stirred. After 15 minutes, 4-nitrobenzylbromide (4.2 mg, 0.009 mmol, 1.20 eq.) was added under nitrogen and the reaction mixture was allowed to stir for more 8 hours at room temperature. After the completion of reaction (according to TLC), the reaction mixture was quenched with ice cold water. The product formed was extracted with CH2Q2 and the combined organic layer was dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and crude product obtained was purified with silica gel (230-400 mesh) chromatography using MeOH: CH2Q2 as eluents to provide pure product 12 (compound 1012) in 18% yield. MS (ESI-TOF) m / z calculated for C44H45N3O8 [M+H]+744.3, found 744.2.

[0138] Example 13: 7-O-(2-nitrobenzyl)-fangchinoline (compound 1013): Fangchinoline (10 mg, 0.017 mmol, 1.00 eq.) was dissolved in dry DMF (O.lmL) under inert atmosphere. Sodium hydride (0.9 mg, 0.039 mmol, 2.50 eq.) was added to solution at -10°C and the suspension was stirred. After 15 minutes, 2-nitrobenzylbromide (4.2 mg, 0.009 mmol, 1.20 eq.) was added under nitrogen and the reaction mixture was allowed to stir for overnight at room temperature. After the completion of reaction (according to TLC), the reaction mixture was quenched with ice cold water. The product formed was extracted with CH2CI2 and the combined organic layer was dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and crude product obtained was purified with silica gel (230-400 mesh) chromatography using MeOH: CH2Q2 as eluents to provide pure product 13 (compound 1013) in 49% yield. MS (ESLTOF) m / z calculated for C44H45N3O8 [M+H]+744.3, found 744.5.

[0139] Example 14: 7-O-(3-methoxybenzyl)-fangchinoline (compound 1014): Fangchinoline (10 mg, 0.017 mmol, 1.00 eq.) was dissolved in dry DMF (O.lmL) under inert atmosphere. Sodium hydride (0.9 mg, 0.039 mmol, 2.50 eq.) was added to solution at -10°C and the suspension was stirred. After 15 minutes, 3-methoxybenzylchhloride (3.1 mg, 0.009 mmol, 1.20 eq.) was added under nitrogen and the reaction mixture was allowed to stir for overnight at room temperature. After the completion of reaction (according to TLC), the reaction mixture was quenched with ice cold water. The product formed was extracted with CH2CI2 and the combined organic layer was dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and crude product obtained was purified with silica gel (230-400 mesh) chromatography using MeOH: CH2Q2 as eluents to provide pure product 14 (compound 1014) in 41% yield. MS (ESLTOF) m / z calculated for C45H48N2O7 [M+H]+729.3, found 729.5.

[0140] BIOLOGICAL EXAMPLES:

[0141] The present invention demonstrates the protective effects of Fen, compound 1001, 1003, 1009 and 1011 against intra cellular infection and shows the potent TLR 4 agonistic activity. The combination of Fen with a suboptimal dose of ciprofloxacin offers a synergistic effect, effectively controlling infection and related symptoms.

[0142] Biological Example 1: Direct toxicity assay for effect of Fangchinoline and its derivative on THP-1 and bacterial cells

[0143] 5xl05cells were seeded per well in 96 well plate, 5mM concentration of each compound from stock plates obtained a final concentration of IpM in each well. Media and THP1 cells only act as a control in triplicate established. The culture plates were incubated at 37°C for 24 hours. After 24 h, 20pl of Presto blue (Thermo scientific) was added to each well. Visible color changes were observed after 4 hours. Whereas, to test the direct effect of Fen on bacteria we performed direct inhibition assay. Briefly, overnight incubated bacterial OD was measured at 600nm and diluted in LB media till 0.01 and incubated the bacteria overnight in the presence of Fen from 50pM to 0.0975 pM. OD was measured at 600nm. (Figure 1)

[0144] Biological Example 2: Salmonella infection and Colony forming Assay

[0145] Bacteria were cultured till the log phase and then sub-cultured in LB media for 3 hours at 37°C (200rpm). The optical density of the culture was then taken at 600 nm for a quick estimation of CFU (1 OD600nm= lxl08CFU / ml; Shigella OD600 = 0.5 ~ 2.5 x 108). THP-1 cells were cultured in RPML1640 supplemented with 10% FBS and 1% penicillin-streptomycin, then differentiated with PMA (50ng / ml). After 24 hours, macrophages were infected with 1 : 10 MOI for 1 hour at 37°C, 5% CO2. Following infection, cells were treated and cultured for 24 hours. Subsequently, cells were lysed in 0.01% Triton-X-100, plated on SS-agar, and incubated at 37°C. Colonies were counted and CFU / ml was calculated. (Figure 2)

[0146] Biological Example 3: HEK-Blue reporter cell Assay:

[0147] In order to understand as to how Fen leads to generation of NO and whether Fen activates TLRs, HEK-Blue reporter cell assay was performed with TLR4, TLR7, TLR8 and hNOD2 (Nucleotide-binding oligomerization domain-containing protein. HEK-Blue cells, expressing human TLR4, TLR7, TLR8, or N0D2 genes along with an inducible SEAP reporter gene, were obtained from InvivoGen. Cultured in DMEM supplemented with FCS, penicillinstreptomycin, and Normocin, reagents were diluted and added to plates. Cells were then added and incubated overnight at 37°C with 5% CO2. SEAP levels were measured using QUANTI-Blue Solution, with results presented as OD at 650nm.

[0148] The results indicated that Fen exhibits TLR4 agonistic activity at a concentration of 2.0 pM and has no effect on Monophosphoryl Lipid A (MPLA) (from Salmonella Minnesota) activity (Figure 3a). Fen did not show agonistic activity to (human) hNOD2, hTLR7 and hTLR8 (a right panel). Further the experiment was carried out using TLR4 inhibitor (TLR4-IN-C34) and it was found that Fcn-induced NO production and antibacterial function are compromised (Figure 3 b-c). It was further found that Fen in combination with TLR4 inhibitor suppressed the expression of Fcn-induced proinflammatory cytokines, IL-6, IL-12, and TNF-alpha, in macrophages (Figure 3 d-f). Further LPS, a known TLR4 agonist was used in order to compare Fen functions in these assays. It was found that Fen induces NO similar to LPS and mitigated the effect in presence of TLR4 inhibitor (Figure 3 g). Fen's effect via MYD88, a downstream of TLR4 was examined. Treating wt. and MYD 88 -deficient BMDMs and BMDMs with Fen revealed compromised antibacterial function, and pro -inflammatory cytokine induction (IL-6, IL-12, TNF-a) without MYD88 signaling (Figure 3 h-j).

[0149] Biological Example 4: Synergistic effect of Fen in enhancement of antibiotic function. To assess Fen's potential to enhance antibiotic efficacy and boost protective immunity against Salmonella, a chronic Salmonella infection model (129SvJ) was used. Fen induces antibacterial Thl immunity via TLR4-MYD88-NO pathway, prompting investigation into its synergy with antibiotics. Combining Fen with low-dose Cipro effectively controlled Salmonella growth. The combination matched high-dose Cipro's effectiveness, rescuing body weight reduction observed with low-dose Cipro alone (Figure 4 a-b). Fcn-Cipro and 1001 and cipro combination suppressed Salmonella growth akin to high-dose Cipro, alleviating shorter colon seen with infection (Figure 4 c-d). Shows that, Fen shows synergistic effect with conventional antibiotic (Ciprofloxacin) to impede Salmonella infections (Figure 5).

[0150] Biological Example 5: Recovery of infected mice by Fangchinoline and its derivative compound 1001 treatment or in combination with Ciprofloxacin in vivo.

[0151] Mice were infected with IxlO7CFU / mice. After 24 hours post-infection (hpi), mice received daily treatments of compound 1001. Body weight was monitored daily, compound 1001 treatment rescued body weight loss, but not at the suboptimal dose. However, combination of compound 1001 with Cipro yielded a response comparable to or more potent than the optimal compound 1001 dose (Figure 5).

[0152] The gastroenteritis model for Salmonella acute infection was established through oral gavage administration. Briefly, mice were starved of food and water for 4 hours and then orally gavaged with 7.5 mg streptomycin to sterilize the gut. After 20 hours mice were again starved of food and water for 4 hours. Overnight culture of Salmonella and further sub cultured for 4 hours were infected in these mice at IxlO7CFU by oral gavage. After the successful establishment of infection, different group of mice received fangchinoline, and combination (Suboptimal doses). Mice were showing no movement when gently fodded with index finger further, mice were euthanized by CO2 as stipulated by IACE protocols. Biological Example 6: Histopathology

[0153] Excised tissues of animal organs were fixed in 10% formalin solution and processed for paraffin embedding. The paraffin blocks were cut into 3-pm-thick sections and then mounted on glass slides. One section from each sample was stained with hematoxylin and eosin. Colon samples were blotted with Haematoxylin and Eosin. Each stained section was analyzed and captured at 20X magnifications. Professional histologist performed Blind assessment and scoring of each section for each sample. (Figure 6)

[0154] Histological analysis revealed that Fen or compound 1001 derivative combined with Ciprofloxacin resulted in reduced inflammation, ulcers, and edema compared to untreated or suboptimal doses of Fen, compound 1001 derivative.

[0155] The compounds of the present invention act as TLR 4 agonists. (Figure 7)

[0156] The present invention discloses the protective effect of Fen and derivatives against Salmonella in-vivo. Fen, and its derivatives synergistically controls infection and associated symptoms in combination with Sub optimal dose of cipro.

Claims

Claim:

1. A compound of chemical formula I along with its pharmaceutically acceptable salts and isomers;FORMULA IWherein:A is a lone pair, or selected from a group comprisingRi and R2 are selected from a group of hydrogen, halogen, cyano, nitro, substituted or unsubstituted haloalkyl, substituted or unsubstituted alkyl and substituted or unsubstituted cycloalkyl; or B(0H)2, NRaRb, , -NRaC(O)Rb, or - NRaS(O)Rb, ORa; Raand Rb, which may be same or different at each occurrence, are independently selected from the group consisting of hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkyl / alkyl, substituted or unsubstituted aryl, substituted or unsubstituted aryl / alkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaryl / alkyl, and substituted or unsubstituted heterocyclyl / alkyl; -C(O)ORa, -C(O)Ra, -C(O)NRa, and -S(O)Ra;2. The compound as claimed in claim 1, as represented by chemical formula II;FORMULA IIWherein:A is a lone pair, or is selected from a group comprising> Ra, which may be same or different at each occurrence, is independently selected from the group consisting of substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted haloalkyl, substituted or unsubstituted cycloalkyl, -C(O)ORa, -C(O)Ra, -C(O)NRa, and -S(O)Ra;R2 may be hydrogen, halogen, cyano, nitro, substituted or unsubstituted haloalkyl, substituted or unsubstituted alkyl and substituted or unsubstituted cycloalkyl; or B(OH)2, NRbRc, -NRbC(O)Rc, or - NRbS(O)Rc;Rb and Rc, which may be same or different at each occurrence, are independently selected from the group consisting of hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkyl / alkyl, substituted or unsubstituted aryl, substituted or unsubstituted aryl / alkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaryl / alkyl, and substitutedor unsubstituted-COO(CH2)nCH3, COCH2CI;n is 6-15.

3. The compounds as claimed in claim 1 are:1) 1001: 16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34- octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)- dibenzenacyclooctaphane-37-yl methanesulfonate;2) 1002: 16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34- octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)- dibenzenacyclooctaphane-37-yl 4-methylbenzenesulfonate;3) 1003: 16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34- octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)- dib enzenacy cl ooctaphane -37-y 1 acetate ;4) 1004: 16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34- octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)- dib enzenacy cl ooctaphane -37-y 1 noni onate;5) 1005: 16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34- octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)- dib enzenacy cl ooctaphane -37-y 1 hexadecyrate;6) 1006: hexadecyl (16,36,54-trimethoxy-12,32-dimethyl- ll,12,13,14,31,32,33,34-octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina- 5(l,3),7(l,4)-dibenzenacyclooctaphane-37-yl) carbonate;7) 1007: 16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34- octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)- dibenzenacyclooctaphane-37-yl 2-(9H-fluoren-9-yl)acetate;8) 1008: 16,36,54-trimethoxy-12,32-dimethyl-ll,12,13,14,31,32,33,34- octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina-5(l,3),7(l,4)- dibenzenacyclooctaphane-37-yl 2,4-difluorobenzoate;9) 1009: 37-(allyloxy)-16,36,54-trimethoxy-12,32-dimethyl- ll,12,13,14,31,32,33,34-octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina- 5 ( 1 , 3 ),7( 1 ,4)-dib enzenacy cl ooctaphane ;10) 1010: 37-butoxy-16,36,54-trimethoxy-12,32-dimethyl- ll,12,13,14,31,32,33,34-octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina- 5 ( 1 , 3 ),7( 1 ,4)-dib enzenacy cl ooctaphane ;11) 1011: 37-(benzyloxy)-16,36,54-trimethoxy-12,32-dimethyl- ll,12,13,14,31,32,33,34-octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina- 5 ( 1 , 3 ), 7 ( 1 ,4) -dib enzenacy cl ooctaphane ;12) 1012: 16,36,54-trimethoxy-12,32-dimethyl-37-((4-nitrobenzyl)oxy)- ll,12,13,14,31,32,33,34-octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina- 5 ( 1 , 3 ), 7 ( 1 ,4) -dib enzenacy cl ooctaphane ;13) 1013: 16,36,54-trimethoxy-12,32-dimethyl-37-((2-nitrobenzyl)oxy)- ll,12,13,14,31,32,33,34-octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina- 5 ( 1 , 3 ), 7 ( 1 ,4) -dib enzenacy cl ooctaphane ;14) 1014: 16,36,54-trimethoxy-37-((3-methoxybenzyl)oxy)-12,32-dimethyl- ll,12,13,14,31,32,33,34-octahydro-2,6-dioxa-l(7,l),3(8,l)-diisoquinolina- 5(1, 3), 7(1 ,4)-dibenzenacyclooctaphane .

4. A method for synthesis of the compounds as claimed in claim 1, comprising the steps of;i. dissolving fangchinoline in anhydrous CH2CI2 or ethyl acetate (EtOAc) or tetrahydrofuran (THF) or acetonitrile (MeCN) or toluene under an inert atmosphere;ii. adding tri ethylamine and the corresponding sulfonyl chlorides or acyl chlorides to the solution of step i at a temperature of 0-5 °C;iii. stirring the reaction mixture obtained from step ii at room temperature and quenching with ice cold water upon completion;iv. extracting the product obtained from step iii with CH2Q2, and drying of the combined organic layer over anhydrous sodium sulfate or magnesium sulfate or calcium chloride (CaCh) or potassium carbonate (K2CO3);v. evaporating the solvent under reduced pressure to yield a crude product; vi. purification of the product of step v by silica gel column chromatography using MeOH: CH2CI2 as eluents, or preparative thin-layer chromatography to obtain pure products.

5. A combination of the compounds as claimed in claim 1, with structurally similar compounds like Fen in conjunction with Fen.

6. A combination of compounds as claimed in claim 1, with fangchinoline (Fen) and ciprofloxacin or enrofloxacin.

7. A composition of the compounds as claimed in claim 1 with pharmaceutically acceptable excipients and adjuvants selected from a group comprising binders such as lactose, microcrystalline cellulose, starch, povidone, and hydroxypropyl methylcellulose (HPMC); diluents like calcium phosphate, mannitol, sorbitol, microcrystalline cellulose, and lactose; disintegrants including croscarmellose sodium, sodium starch glycolate, crospovidone, and starch; lubricants such as magnesium stearate, stearic acid, sodium stearyl fumarate, and talc; glidants like colloidal silicon dioxide and talc; preservatives such as benzalkonium chloride, benzoic acid, potassium sorbate, and sodium benzoate; stabilizers like citric acid, ascorbic acid, tocopherols, and EDTA; sweeteners including aspartame, sucralose, sorbitol, and xylitol; flavors and colors like natural and artificial flavoring agents, FD&C colorants, and caramel color; and solubilizers such as polyethylene glycol (PEG), polysorbate 80, and propylene glycol.

8. The adjuvants of the composition as claimed in claim 7, selected from a group of aluminum salts (e.g., aluminum hydroxide, aluminum phosphate), oil-in-water emulsions like squalene-based adjuvants (e.g., MF59, AS03), cytokine modulators such as granulocyte-macrophage colony-stimulating factor (GM-CSF), Toll-Like Receptor (TLR) agonists like monophosphoryl lipid A (MPL) and CpG oligonucleotides, polysaccharides including chitosan and dextran, lipid-based adjuvants such as liposomes and phospholipids, saponins like QS-21, and polymeric adjuvants such as poly(lactic-co-glycolic acid) (PLGA) and polyinosinic-polycytidylic acid (poly-IC).

9. A composition comprising the combination as claimed in claim 5 and 6, along with pharmaceutically acceptable excipients and adjuvants.

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