A graphene-anticancer peptide complex with nanohole

Abstract

The invention relates to the biocompatible complex formed by binding a pyrene- methanol-bonded therapeutic peptide (Cycas Revoluta anticancer peptide 1, Cr-ACP1) to the quadrilateral graphene nanohole surface with a defective structure, and the synthesis method of this complex. With the therapeutic peptide attached to the biocompatible complex, treatment is provided to healthy tissues with minimal side effects, and the transport of said complex to the cancer cell occurs in a controlled manner.

Description

[0001] DESCRIPTION

[0002] A GRAPHENE-ANTICANCER PEPTIDE COMPLEX WITH NANOHOLE STRUCTURE

[0003] Technical Field of the Invention

[0004] The invention relates to the biocompatible complex formed by binding a pyrene- methanol-bonded therapeutic peptide (Cycas Revoluta anticancer peptide 1 , Cr-ACP1 ) to the quadrilateral graphene nanohole surface with a defective structure, and the synthesis method of this complex. With the therapeutic peptide attached to the biocompatible complex, treatment is provided to healthy tissues with minimal side effects, and the transport of said complex to the cancer cell occurs in a controlled manner.

[0005] State of the Art

[0006] Cancer is defined as a broad group of diseases that can begin in almost any organ or tissue of the body when abnormal cells grow uncontrollably, exceed normal boundaries, invade adjacent parts of the body, and / or spread to other organs. According to the World Health Organization, cancer is the second leading cause of death worldwide, with an estimated 9.6 million deaths, or one in six deaths, in 2018.

[0007] Colorectal cancer, one of the types of cancer, is a disease in which cells in the colon or rectum begin to grow uncontrollably. Most colorectal cancers begin as a growth on the inner lining of the colon or rectum, called a polyp, and if cancer forms in a polyp, it can grow into the wall of the colon or rectum over time, causing cancer. In addition, while cancer cells are in the wall, they can grow into blood vessels or lymph vessels, and from there they travel to nearby lymph nodes or distant parts of the body, causing metastasis. Estimates by the American Cancer Society for the number of colorectal cancer cases in the United States for 2022 indicate that there will be 106,180 new cases of colon cancer and 44,850 new cases of rectal cancer. Therefore, research on alternative treatments for colon cancer is gaining importance day by day.

[0001] .

[0008] Common treatment methods for colon cancer include surgery, chemotherapy and anticancer drugs. However, these treatment methods have many side effects, such as nausea, hair loss, and feeling of fatigue, and they also negatively affect healthy cells. There are also some plant-based therapeutic peptides for use in the treatment of colon cancer. It is known that in the state of the art, Cr-ACP1 (Cycas Revoluta anticancer peptide 1 ), obtained by purification from the seeds of the Cycas Revoluta (Palm Fern) plant available, has harmful effects against human epidermoid cancer (Hep2) and colon carcinoma cells (HCT 15) and that it inhibits the proliferation of cancer cells and induces apoptosis as a result of binding directly to DNA by disrupting the nucleosome structure [2], In addition, there are also drugs that provide targeted therapy, and the delivery of these drugs to cancer cells occurs through biocompatible complexes. In the state of the art, various nanomaterials such as nanoparticles, polymers, carbon nanotubes and graphene are widely used for controlled drug delivery and release in order to transport drugs. However, the surface binding and transport capacity of the transported peptide or drug is insufficient. In addition, transported peptides or chemically derived drugs show higher levels of toxicity than herbal anticancer agents. Therefore, there is no biocompatible complex that enables nanocarriers to reach tumours more selectively than free agents, especially by providing controlled peptide agent transport to the colon cancer cell, with increased permeability and retention effects, while showing low toxicity to healthy tissues and high therapeutic effectiveness to tumour tissues.

[0009] Due to reasons such as the limitations and inadequacies of the solutions in the state of the art, lack of a biocompatible complex that provides controlled peptide transport to cancer cells, especially colon cancer, drugs or peptides used in cancer treatment having low activity, specificity, affinity and having insufficient ability to penetrate cell membranes, and the damage to healthy cells during cancer treatment, it has become necessary to provide a biocompatible complex that provides controlled peptide transport to cancer cells, especially colon cancer.

[0010] Brief Description and Aims of the Invention

[0011] In the invention, the biocompatible complex formed by bonding Cr-ACP1 with the amino acid sequence SEQ ID NO:1 bonded to pyrene-methanol to the quadrilateral graphene nanohole surface with a defective structure and the synthesis method of this complex are described.

[0012] The aim of the invention is to create a biocompatible complex comprising pyrene methanol-Cr-ACP1 for use in cancer treatment. In the invention, the apoptosis of cancerous cells is induced by means of the Cr-ACP1 in the complex, and highly ordered IT— IT stacks are formed by conjugating pyrene-methanol to Cr-ACP1 . Therefore, the peptide is transported to the cancer cell at high efficiency.

[0013] Another aim of the invention is to create a complex with defective quadrilateral graphene nanohole surface. By means of the defective structure of said surface, the affinity and carrying capacity of Cr-ACP1 is increased.

[0014] Another aim of the invention is to provide cancer treatment with a herbal peptide agent as an alternative to cancer drugs. With the use of herbal peptide agents, therapy functions are performed with minimal side effects on healthy tissues.

[0015] Another aim of the invention is to ensure the controlled transport of the herbal peptide to the cancer cell with the biocompatible complex created.

[0016] Description of Drawings

[0017] Figure 1 : Mean square displacement plot of graphene-peptide complex

[0018] Figure 2: Analysis of Inertial (Rotation) radius for peptides

[0019] Figure 3: Graph of the effect of time-dependent change of ambient temperature on complex stability

[0020] Detailed Description of the Invention

[0021] In the invention, the biocompatible complex formed by bonding Cr-ACP1 with the amino acid sequence SEQ ID NO:1 bonded to pyrene-methanol to the quadrilateral graphene nanohole surface with a defective structure and the synthesis method of this complex are described.

[0022] The Cr-ACP1 peptide with the amino acid sequence SEQ ID NO:1 mentioned in the invention was conjugated with pyrene methanol, by means of this conjugation, highly ordered TT-TT and h-n interactions are formed and the transport of the peptide to the cancer cell occurs at high efficiency. In addition, the unit cell side length of said quadrilateral graphene nanohole surface is 18 A and the quadrilateral nanohole side length is 6 A, and by means of the nanohole form of the surface, the controlled binding, transport and controlled release capacity of the peptide mentioned in the invention to the surface is increased. Synthesis method of the biocompatible complex created for use in cancer treatment comprises the process steps of: i. synthesising the nanographene using acetylene gas in a chemical vapour deposition reactor under temperature of 900-1200°C and a pressure of 0.5-2 atm, ii. creating the guadrilateral graphene nano hole surface as a result of cooling the synthesized nano graphene at (-143.15) - (-133.15) °C and creating 6:18 zigzag pattern nano holes with an argon ion beam at a dose of 1 x 1 o13Ar+ / cm2at 3 keV, iii. synthesising Cr-ACP1 peptide with amino acid sequence SEQ ID NO:1 in solid phase in dimethylformamide with microwave support, iv. obtaining methanol-peptide complex from pyrene as a result of the conjugation of pyrene methanol and Cr-ACP1 with the amino acid sequence SEQ ID NO:1 by esterification reaction in dichloromethane solvent, v. after adding pure water, 1 -3% sodium chloride (NaCI) and 5-8% synthesized nano graphene into the reactor, treating the components in the reactor in an ultrasonic bath at 25-40°C for 30-45 minutes and ensuring the homogeneous distribution of nano graphene in the mixture, vi. adding 20-30% pyrene methanol-peptide complex to the mixture in which the resulting nano graphene is homogeneously dispersed and keeping the mixture stirring at 25°C for 15-20 hours, and vii. separating the solid phase, which will be used as a drug distribution system, by filtering the mixture to which the resulting pyrene-methanol-peptide complex is added.

[0023] Figure 1 shows the mean square displacement or deviation of the position of the peptide relative to a reference location on the graphene surface. In the graph, the lines marked as peptide 1 , 2, 3, 4 represent the same peptide (Cr-ACP1 with the amino acid sequence SEQ ID NO: 1 bound to pyrene-methanol) and are numbered differently in order to observe the deviation of the peptide according to different reference positions. By mean square displacement analysis, it was determined that the amount of displacement was limited by the length of graphene. Figure 2 shows the analysis of the inertia radius and was carried out to check that the graphene surface with a defective structure provides sufficient space for peptide placement. The graph in Figure 2 shows the radius of inertia, or the measure of the distance between the centre of mass of the peptide and both its terminals. In addition, the results of the analysis performed to examine the effect of timedependent change of ambient temperature on the stability of the complex are shown in Figure 3. It was determined that during the analysis period (500ps), the peptide- graphene complex could remain stable under the time-dependent change of the analysis ambient temperature and the conditions of the complex at room temperature (298K).

[0024] REFERENCES

[0025] 1. Colorectal (colon) cancer: Symptoms, diagnosis, treatments. Cleveland Clinic, (n.d.). Retrieved October 18, 2022, from https: / / my.clevelandclinic.org / health / diseases / 14501 -colorectal-colon-cancer 2. Mandal SM;Migliolo L;Das S;Mandal M;Franco OL;Hazra TK; (n.d.). Identification and characterization of a bactericidal and proapoptotic peptide from Cycas revoluta seeds with DNA binding properties. Journal of cellular biochemistry. Retrieved September 27, 2022, from https: / / pubmed.ncbi.nlm.nih.gov / 21882228 /

Claims

CLAIMS1. Synthesis method of a biocompatible complex comprising plant peptide, comprising the process steps of: i. synthesising the nanographene using acetylene gas in a chemical vapour deposition reactor under temperature of 900-1200°C and a pressure of 0.5-2 atm, ii. creating the guadrilateral graphene nano hole surface as a result of cooling the synthesised nano graphene at (-143.15)- (-133.15) °C and creating 6:18 zigzag pattern nano holes with an argon ion beam at a dose of 1 x13Ar+ / cm2at 3 keV, iii. synthesising Cr-ACP1 with amino acid sequence SEQ ID NO:1 in solid phase in dimethylformamide with microwave support, iv. obtaining methanol-peptide complex from pyrene as a result of the conjugation of pyrene methanol and Cr-ACP1 with the amino acid sequence SEQ ID NO:1 by esterification reaction in dichloromethane solvent, v. after adding pure water, 1 -3% sodium chloride (NaCI) and 5-8% synthesized nano graphene into the reactor, treating the components in the reactor in an ultrasonic bath at 25-40°C for 30-45 minutes and ensuring the homogeneous distribution of nano graphene in the mixture, vi. adding 20-30% pyrene methanol-peptide complex to the mixture in which the resulting nano graphene is homogeneously dispersed and keeping the mixture stirring at 25°C for 15-20 hours, and vii. separating the solid phase, which will be used as a drug distribution system, by filtering the Mixtures to which the resulting pyrene- methanol-peptide complex is added.

2. Synthesis method of a biocompatible complex comprising herbal peptide according to claim 1 for use in cancer treatment.

3. A method according to claim 2, wherein said cancer is colon cancer.

4. A method according to claim 1 , wherein the peptide mentioned in the process step (iii) is the Cr-ACP1 peptide with the amino acid sequence SEQ ID NO:1.

5. A method according to claim 1 , wherein said nanographene is synthesised under 900-1200°C temperature and 0.5-2 atm pressure.A biocompatible complex with a defective structure obtained by a method according to stem 1 . A biocompatible complex created for use in cancer treatment, wherein it is Cr- ACP1 with the amino acid sequence SEQ ID NO:1 , conjugated with 20-30% pyrene-methanol, attached to the quadrilateral graphene nanohole surface with unit cell side length of 18 A and side length of 6 A. A biocompatible complex comprising plant peptides, wherein it is Cr-ACP1 with the amino acid sequence SEQ ID NO:1 , conjugated with 20-30% pyrenemethanol, attached to the quadrilateral graphene nanohole surface with unit cell side length of 18 A and side length of 6 A. Formulation comprising a biocompatible complex according to any of the preceding claims. A formulation according to Claim 9; wherein it is in injection form.

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