Chlorogenic acid for injection is the world's first class I new anti-cancer drug independently developed by Jiuzhang Biotech, and belongs to anti-tumor immune small molecule natural medicines. Jiuzhang Biotech has systematically researched and developed chlorogenic acid since 2000, and has secured the clinical approval documents for chlorogenic acid drug substance and chlorogenic acid for injection in August 2013 (approval numbers: 2013L01855 and 2013L01856). In December 2016, the Phase I/II Clinical Study on Innovative Class I New Drug Chlorogenic Acid for Injection was listed by the National Health and Family Planning Commission and the Ministry of Science and Technology in the national science and technology major new drug discovery projects in 13th Five-year plan, which was also a major science and technology support project in Sichuan Province.

1. General

Nature is a complex organism made up of a variety of interdependent and interinhibitive substances. Sunlight, water, microorganisms, plants and animals of all kinds survive in an interinhibitive environment. The microcosm, like the macrocosm, is also made up of a variety of interdependent and interinhibitive substances which form a complex and diverse objective environment. The universe itself has problems when it was formed, and the ways and means or the keys to solve all problems exist in nature. What human beings have to do is to maintain the development track of the objective nature rule, and if this rule is broken, it will inevitably produce “environmental problems”. Nature is an inexhaustible treasure trove, and any existence (substance) is a necessity in the objective rule, and is the constraint point in the cycle of nature, which support, control and maintain the balance of nature. From the perspective of objective facts, drugs that are artificially created and synthesized often have serious environmental problems (toxic side effects). And all of the century-old “good medicines” are derived from nature’s gifts, such as penicillin, aspirin, and metformin and so on.

Sichuan Jiuzhang Biological Science and Technology Co., Ltd. (hereinafter called Jiuzhang Biotech or the Company), following the guiding principle of the objective nature rule, has been focusing on the innovative research of small molecule compounds originated from natural plants for over 20 years. Based on the Company’s original extraction, separation and purification technology platform for medicinal natural products, nearly 10 natural medicines, represented by chlorogenic acid, have been selected for natural drug development. And it is the first time in the world that chlorogenic acid, which widely exists in plants, is used for the research and development of the treatment of major diseases (cancers, neurological disorders, autoimmune disorders, etc.).

2. The Project of Chlorogenic Acid for Injection

Chlorogenic Acid for Injection is the world’s first new anti-tumor drug, which is a major breakthrough innovation in the field of tumor therapy, especially in inducing cancer cell differentiation (a new strategy to turn enemies into friends) and metabolic regulation. In 2016, the project was included by the National Health Planning Commission of China and the Ministry of Science and Technology in the 13th Five-Year Plan of the People’s Republic of China as a major new drug creation and production project, and passed the project acceptance at the end of 2020.

The Project has the following characteristics:

 (1) Natural source: Chlorogenic Acid for Injection is a single-molecule active compound extracted from plants, with special physicochemical properties; it is difficult to synthesize large quantities of high-purity chlorogenic acid API to meet the requirements for drug development with the existing chemical technology;

 (2) Safety: Data from the Phase I clinical study and Phase II clinical study (phased data) of Chlorogenic Acid for Injection shows an excellent safety: except the development of muscle hardness at the site of injection, no other drug-related adverse reactions of any kind occurred;

 (3) High efficiency: on the basis of improving the quality of survival of patients with advanced tumors, the effect of controlling tumor progression is obvious, effectively prolonging the survival of patients with advanced tumors;

 (4) Unique mechanism of action: Mechanism of action and clinical studies have confirmed that chlorogenic acid has a clear target point, is a natural small molecule anti-cancer drug with multi-channel, multi-layer and wide anti-tumor spectrum targeting cell mitochondria, and has the effect of inducing differentiation (reduction of malignancy or transformation into benign) and metabolic checkpoint regulation (mtACAT1) in tumor cells;

 (5) Broad-spectrum anti-cancer: It has significant efficacy against many types of malignant tumors such as lung squamous carcinoma, glioma, esophageal squamous carcinoma, cervical squamous carcinoma, oral cavity squamous carcinoma, prostate cancer, etc. It has a broad anti-tumor spectrum and has significant clinical needs;

 (6) It can be applied alone or synergistically and for sensitization: it can be used synergistically with a variety of chemotherapeutic, targeted, and immunotherapeutic agents, resulting in a significant increase in OS (overall survival) (the use of Chlorogenic Acid for Injection restores drug sensitivity in cases of resistance to other drug treatments);

 (7) Wide range of application: Based on the biological active pharmaceutical characteristics of chlorogenic acid with clear target point and multi-pathway, it has been confirmed that, in addition to anti-tumor treatment and relief of cancer pain, Chlorogenic Acid for Injection also shows good efficacy on non-controllable inflammation as well as metabolic disorders, and it can be applied to the treatment of major diseases such as hyperlipidemia, cirrhosis of the liver, Alzheimer’s disease and so on. It has a broad space for sustainable development, and is expected to become a large variety of drug in China;

 (8) Chlorogenic Acid for Injection results from the original research of the world’s first new drug, the patent framework is rigorous, the API technology bottleneck is high, and at present only Jiuzhang Biotech master the world’s key technologies, while other institutions or companies are involved in researching the mechanisms of treating diseases;

 (9) At present, the risk of the project has been greatly released, and the perspective of druggability is strong:

① Good safety: the results of phase I clinical trials completed in Beijing Cancer Hospital and Beijing Shijitan Hospital and the data of the Chlorogenic Acid for Injection subjects in phase II/III clinical trials currently underway indicate that the Chlorogenic Acid for Injection is safe and well-tolerated, with rapid metabolism in the human body and no accumulation of toxicity.

② Apparent therapeutic advantages: Chlorogenic Acid for Injection has shown great therapeutic advantages in the clinical trials for the treatment of recurrent high-grade gliomas, which can significantly improve the quality of life of patients with advanced tumors and prolong the survival period of patients (the globally recognized median survival period of patients with multiple recurrence of grade Ⅳ gliomas is only 3-4.6 months, but the median survival period of the subjects treated with Chlorogenic Acid for Injection (Phase I effective dose group) is 21.4 months, far exceeding the international standard. From the preliminary interim statistics of the current phase II/III clinical trials, the mortality rate of subjects in the chlorogenic acid group is significantly lower than that of the CCNU (Lomustine, positive control drug) group, and the survival period is also better than that of the CCNU group (with the prolongation of the treatment time, the survival advantage of the chlorogenic acid group will be even more obvious).

③ Single-species multi-pipeline: Chlorogenic Acid for Injection has broad-spectrum anti-cancer pharmaceutical properties and can be applied to a wide range of tumor treatments; according to the exploration of preliminary clinical trials and basic pharmacodynamic studies, Chlorogenic Acid for Injection may also be suitable for the treatment of other diseases, such as metabolic diseases.

3.  Intellectual Property Rights and Project Initiation

3.1 Intellectual property rights

After years of efforts, Jiuzhang Biotech has initially established an international and domestic patent protection network for chlorogenic acid, involving crystal structure, preparation process, route of administration, dosage form design and indications, etc., and can realize comprehensive protection of the main commercial interests of chlorogenic acid-related products worldwide.

As of December 31, 2023, Jiuzhang Biotech has obtained 52 domestic patents, including 51 invention patents and 1 utility model patent, and has obtained 22 foreign patents, invention patents, including 14 patents in the United States, 4 patents in Europe, 3 patents in Japan and 1 patent in South Korea.

All patents are exclusively owned by Jiuzhang Biotech, with clear ownership, and no joint and authorized use licenses, which ensure the advantageous leading position of chlorogenic acid-related products in the market competition in the future.

Meanwhile, the Company has registered and obtained 15 valid trademark rights, including words and designs, in various important fields such as drugs for human use, and APIs.

3.2 Projects supported for initiation

l  2014 Sichuan Science and Technology Support Program.

l “PhaseⅠ/Ⅱ Clinical Study of First Creation Class I New Drug Chlorogenic Acid for Injection” 2016ZX09101017 (This project is the only one titled as “first of its kind” in the “Creation of Major New Drugs” of the “13^th Five-Year Plan” of China’s major scientific and technological special projects so far).

l   Listed as a key project of the governor of Sichuan Province in 2017

l  2017 Sichuan Provincial Major Science and Technology Special Project “Phase II Clinical Study of Class 1 Natural Small-molecule Immunity Innovative Drug Chlorogenic Acid for Injection in the Treatment of Brain Glioma”

l     2019 Chengdu Biomedical Post-subsidy Funding Project “Phase II Clinical Study of Chlorogenic acid for Injection (Chemical Drug Class 1.2, approval No. 2013L01855)”

l  2020 Major Science and Technology Innovation Project of Chengdu Science and Technology Bureau: “Phase II/III Clinical Study of Chlorogenic Acid for Injection, a New Class 1 Drug for Malignant Glioma”.

4.Study on Mechanism of Action of Chlorogenic Acid for Injection

The mechanism study team of the Institute of Materia Medica of the Chinese Academy of Medical Sciences, clinical and trial study teams of Beijing Tiantan Hospital (Phase I clinical trial completed in Beijing Shijitan Hospital), Xi'an Tangdu Hospital, Tianjin Huanhu Hospital, the Second People’s Hospital of Shenzhen and Chongqing Cancer Hospital together with the Jiuzhang Biotech team have conducted in-depth study on the anti-cancer mechanism of chlorogenic acid and performed confirmatory study based on biological samples from Phase I/II clinical study.

The results of the currently public mechanism of action studies are summarized as below:

1). Acting on the mitochondria of tumor cells, it is expected to be the world’s first cancer cell differentiation inducer for solid tumors

(1) Chlorogenic acid is an effective differentiation inducer for many kinds of solid tumor cells: the mechanism of action can be briefly described as chlorogenic acid acts on mitochondria, regulates ATP, increases small ubiquitylated c-Myc in tumor tissues through SUMO1 protein, reduces phosphorylated c-Myc, reduces miR-17 family, increases p21, induces cell differentiation, extends the time of doubling, slows down the growth of solid tumors, and reduces metastasis and invasive power. On the other hand, by inducing cell differentiation, reducing PD-L1 expression in cancer cells, protecting immune effector cells in the tumor microenvironment, improving the immune microenvironment of tumors, and ultimately inhibiting or destroying tumors, thus realize both symptomatic and curative treatment.

 (2) Regulation of acetyl coenzyme A acetyltransferase 1 (mtACAT1): Using the natural compound target angling technique based on photoaffinity labeling, combined with protein microarray technology, we searched for and discovered the potential target of chlorogenic acid, acetyl coenzyme A acetyltransferase 1 (mtACAT1). The experimental results showed that the tumor growth of mouse melanoma cells with knockdown of acetyl coenzyme A acetyltransferase 1 gene was significantly slowed down after inoculation into the subcutaneous of mice, and T lymphocytes infiltrated into the tumor tissues were significantly increased, suggesting that the growth and proliferation of the tumor cells were significantly related to mtACAT1. It has been shown that acetyl coenzyme A acetyltransferase 1 can be phosphorylated by some cell growth factor receptor kinases in vivo, and the phosphorylated acetyl coenzyme A acetyltransferase 1 acquires the ability to promote cell proliferation, thus inducing tumors or promoting the proliferation of tumor cells and tumor growth. Our study shows that chlorogenic acid affects the three-dimensional conformation of mtACAT1 through an allosteric effect, thereby inhibiting the phosphorylation of acetyl-coenzyme A acetyltransferase 1 tyrosine 407, and therefore inhibiting tumor cell growth, proliferation, and invasion, which has a clear anti-tumor effect.

Validation of CHA interaction with mtACAT1 by cryo-electron microscopy at Tsinghua University

Under normal conditions, acetyl coenzyme A acetyltransferase 1 (mtACAT1) transfers its own acetyl group to other proteins, acetylates other proteins, and regulates the activity of other proteins, especially inducible proteins including inducible enzymes, in order to maintain the balance of metabolism, immunity, and other functions in the body. However, with age increased, various functions of the body decline, acetyl coenzyme A acetyltransferase 1 activity are also reduced, and the function of acetylation of other proteins is reduced. On the one hand, the ability to regulate inducible enzymes (often belong to the disease-causing enzymes such as COX2, etc.) is reduced, resulting in chronic diseases, and in the central nervous system will cause Alzheimer’s disease, Parkinson’s disease and so on. On the other hand, there are some biologically active substances in the organism that can pass through the blood-brain barrier and blood-testis barrier only after they are acetylated, which can play the roles of nourishing brain tissues and increasing sperm vitality. Once the activity of acetyl coenzyme A acetyltransferase 1 decreases, brain cognitive disorders and sexual dysfunctions will develop over time. Therefore, mtACAT1 is closely related to the aging of the body and the development of major chronic diseases.

2). Chlorogenic acid promotes macrophage M1-type polarization, inhibits macrophage M2-type polarization, improves the tumor microenvironment, affects the interaction between macrophages and tumor cells, and inhibits the growth of tumor cells. At the same time, chlorogenic acid can activate macrophages and enhance the ability of macrophages to phagocytose foreign particles and tumor cells.

Macrophages are microglia in the skull. Under normal circumstances, microglia are dominated by the M1 phenotype, which plays a role in removing dead cell debris; after becoming gliomas, the M2 phenotype is dominant, which does not play a role in removing the tumor but promotes tumor growth. Chlorogenic acid can directly inhibit the M2 phenotype of intracranial microglia and activate and strengthen the M1 phenotype of microglia through the polarization of macrophages, which can play a therapeutic role in the treatment of glioma.

Extracellular deposits of misfolded β⁃amyloid (Aβ), deposits of hyperphosphorylated tau proteins, and widespread neuronal apoptosis are causative factors in Alzheimer’s disease (AD). Normally, these two groups of proteins, as well as apoptotic cells, are cleared by macrophages. However, as the body is unable to metabolize these proteins as age increased, toxic substances continue to be deposited that can lead to AD. Chlorogenic acid can have a therapeutic effect on AD by reducing the number of degenerated and necrotic microglial cells in the skull, or by activating them to become functional microglial cells, which can remove these two proteins in time.

Summary: The above mechanism study reveals that chlorogenic acid can induce tumor suppression by differentiation, reprogramming of cellular metabolism, regulating the interaction between tumor cells and immune cells in the tumor microenvironment, and anti-tumor immunity.

The following articles related to the mechanism have been published so far:

     •    Institute of Materia Medica, Chinese Academy of Medical Sciences: Chlorogenic acid inhibits glioblastoma growth through repolarizating macrophage from M2 to M1 phenotype;

•  Institute of Materia Medica, Chinese Academy of Medical Sciences: Chlorogenic acid effectively treats cancers through induction of cancer cell differentiation;

•    Institute of Materia Medica, Chinese Academy of Medical Sciences: Isolation and identification of human metabolites from a novel anti-tumor candidate drug 5-chlorogenic acid injection by HPLC-HRMS/MSn and HPLC-SPE-NMR;

•    Beijing Tiantan Hospital of Capital Medical University: First-in-human immunomodulation mechanism research of chlorogenic acid in patients with recurrent high-grade glioma;

•    Beijing Cancer Hospital: Development of an LC-MS/MS method for quantitative analysis of Chlorogenic acid in human plasma and its application to a pharmacokinetic study in Chinese patients with advanced solid tumor

•    Institute of Materia Medica, Chinese Academy of Medical Sciences: Chlorogenic acid inhibits esophageal squamous cell carcinoma growth in vitro and in vivo by downregulating the expression of BMI1 and SOX2

•    Institute of Materia Medica, Chinese Academy of Medical Sciences: Integrated proteomic and metabolomic profiling the global response of rat glioma model by temozolomide treatment

•    Institute of Materia Medica, Chinese Academy of Medical Sciences: Tumor-selective lipopolyplex encapsulated small active RNA hampers colorectal cancer growth in vitro and in orthotopic murine

•    Institute of Materia Medica, Chinese Academy of Medical Sciences: Self-assembling HA/PEI/dsRNA-p21 ternary complexes for CD44 mediated small active RNA delivery to colorectal cancer

•    Institute of Materia Medica, Chinese Academy of Medical Sciences: Chlorogenic acid inhibits PD-L1 expression in esophageal cancer cells via the IFN-γ signaling pathway

•    Institute of Materia Medica, Chinese Academy of Medical Sciences: Chlorogenic acid inhibits clone formation and invasive metastasis of esophageal cancer cells through down-regulation of Notch1 expression

•    Institute of Materia Medica, Chinese Academy of Medical Sciences: Progress in the study of the pharmacological effects and mechanism of Chlorogenic acid

•    Institute of Materia Medica, Chinese Academy of Medical Sciences: Characterization of Chlorogenic Acid as a Two-Photon Fluorogenic Probe that Regulates Glycolysis in Tumor Cells under Hypoxia

•   Institute of Materia Medica, Chinese Academy of Medical Sciences: Chlorogenic Acid Induced Neuroblastoma Cells Differentiation via the ACAT1-TPK1-PDH Pathway

•    Institute of Materia Medica, Chinese Academy of Medical Sciences: Cancer Differentiation Inducer Chlorogenic Acid Suppresses PD-L1 Expression and Boosts Anti-tumor Immunity of PD-1 Antibody

•   Beijing Tiantan Hospital of Capital Medical University: Phase I study of chlorogenic acid injection for recurrent high-grade glioma with long-term follow-up

•   Chinese Academy of Medical Sciences: Target fishing and mechanistic insights of the natural anticancer drug candidate chlorogenic acid

5. Progress in Clinical Trials