Synopsis

This is a drug discovery on Haruan Channa striatus, and a proprietary Haruan Traditional Extract (HTE) has been established to contain similar biochemical composition, and sustained the biomedical properties of Haruan. Furthermore, the nano-particles HTE (nHTE) tagged with Tecnitium (99mTc) was traced by gamma-camera in circulatory system entering internal organs and passing blood-brain-barrier into brain tissue in rats, supporting the PC 12 cell-lines research (rats phaechromocytoma cells) demonstrating pharmaco-physiological ability of nHTE as therapeutic agent improving cognitive function and neuroregenerative properties. Therefore, the main objective of this R & D is to establish HTE (a natural product) as therapeutic agent complete with standardisation, certificate-of-analysis (COA), producing prototypes of the OTC products and clinical batches at a GMP facility.


 Introduction

This project is first started in 1985, to discover the potential of Haruan Channa striatus as an undisputed ethnopharmacology, as remedy for wound healing for newly given birth young mothers. Haruan is indegenous to Malaysia, and this freshwater, air breathing and carnivourous fish is belong to family Channidae together with Toman C micropeltes and Bujuk C lucius. There are 30 species in the family, and various different species of the family are found in Pakistan, Indian Sub-continent, ASEAN Countries (Myanmar, Thailand, Laos, Vietnam, Cambodia, the Philippines, Brunei, Indonesia and Singapore) and Southern China (Mohsin and Ambak, 1983). Haruan which is also known as snake head fish, normally prefer shallow stagnant or slow running water or low conductivity, relatively high dissolve oxygen (DO), and neutral pH with little sun light directly on water surface. As carnivorous, Haruan eating smaller fish, amphibian, reptiles as well as birds, and a monogomous with good parental care to the young.

This whole activity is set as an agro-industrial initiative, to commercialising a Drug Discovery Research of the local ethno-pharmacology Haruan C striatus ‘remedy for wound healing’. The activities involves commercial scale farming of the indigenous species, Haruan C striatus, and post-harvest processing into a globally available line of OTC and pharmaceutical products. This initiative is first of all is to produce the active compound(s) in Haruan C striatus, by processing it into Haruan Traditional Extract (HTE), based on an established proprietary extraction protocol, and it is our Trade Secret. The HTE is the bioactive and therapeutic agent in formulation of all the products. As expected, HTE sustained all biochemical profile of Haruan C striatus a traditional remedy to help patients heal from post surgical wounds and internal injuries with a good amino and fatty acid profile, necessary in facilitating wound healing. HTE is also having high percentages of Glycine and Cystine, as well as Arachidonic (AA; 20:4ω6) and Docosahexaenoic Acid (DHA; 22:6ω3). Furthermore, In Vitro as well as In Vivo animal models shows HTE exhibits the following biomedical activities namely anti-nociceptive, anti-inflammatory, antimicrobial, inhibit cancer cell lines, inducing stem cell proliferation, stimulating platelet aggregation and enhancing cognitive functioning.

To comprehend, the whole project are divided into FOUR section:

1. BIOLOGICAL INFORMATION: to include distribution, natural habitat, ecological, genetic background, behaviour and feeding habits. These are the basis of farming activities, feed production and the SOP. With complete information, it is easier to farm Haruan according to specification and to produce enough numbers as required. Furthermore, primers and DNA sequencing on C striatus had been done for identification and breeding purposes (Rahim, et al., 2009)

2. BIOCHEMICAL ANALYSIS: proximate analysis, mineral composition, antioxidant, amino acid and fatty acids profile of Haruan C striatus was compiled to elastrating the physiological goodness of Haruan as health food, and indicating the supplies of basic unit for healing, growth and tissue repair (Mat Jais, et al., 1994)

3. BIOMEDICAL PROPERTIES: wound healing, anti-pain, anti-inflammation, anti-bacteria, anti-cancer, induce blood clotting, induce platelet aggregation, imrpopve cognitive function and neuroregenaration. All the experimentation were using either liquid HTE (lHTE) or nano-particles HTE (nHTE), and both are exhibiting strong biomeical properties (Mat Jais, 2007)

4. BIOMEDICAL PRODUCTS: there are four groups of OTC products namely personal cares, health foods, health spa and medicated cream has been formulated. Some of these especially tonic, softgel capsule and medicated cream will be made into clinical batches for clinical trials Mat Jais, 2007)

All the products are having the aqeous extract or HTE as therpeutic agent that has shown a promising biomedical properties containing antioxidant, wound healing, anti-imflammation, anti-cancer (leukemia and colon), anti-bacetria (E.coli, S typhimurium, S aureus and H pylori; but not inhibiting Lactobacillus case), anti-nociceptive, improving cognitive function and neuroregeneration in rats (Che Ku Dahlan, et al., 2010; Mat Jais, 2007; Mat Jais, et al., 1997; Mat Jais, et al., 1998; Somchit, et al., 2004; Zakaria, et al., 2004;a; 2004b; 2004c and 2005a, b, 2005c; Zuraini, et al., 2006). A Utility Innovation patent (UI 9088631 – process for preparation of fish Haruan Channa striatus extract having pharmaceutical function) has been awardedin 2009, and now the main objective here is to establish and promote Haruan Traditional Extract (HTE) as therapeutic agent for OTC and pharmaceutical products. The project is at a pre-clinical to establsih Standardisation, Certificate-Of-Analysis (COA) and to produce Prototypes at a GMP facility on an outsourcing consignment.

There are three different forms of this in-water proprietary HTE (liquid or lHTE; solid or sHTE and nano-particles or nHTE). All of these HTE containing similar biochemical composition and maintained all the biomedical properties of Haruan as therapeutic agent for wound healing and anti-pain (Mat Jais, et al., 1994; Mat Jais, 2007). With Gamma Camera experimentatiion, where nHTE (the nano-particles) is tagged with radio-leblled Tecnitium (99mTc), given orally to rats, to trace the movement in circulatory system entering internal organs and passing blood-brain-barrier into brain tissue. This experimentation is also carried out using PC 12 cell-lines (rats phaechromocytoma cells that having growth factor receptor on the surface) to demonstrate pharmacological and physiological ability of nHTE as therapeutic agent in improving cognitive function and neuroregenerative properties that comprehend pre-clinical works.


 References

1. Dambisya, YM.; TL.Lee and AM.Mat Jais, 1997. Analgesic Properties of Extracts from Snakehead Fish, Channa striatus, Haruan. Presented at the 8th General Scientific Meeting of the Singapore Society of Anaesthesiology, Singapore, pp 41

2. Dambisya, YM.; TL.Lee and AM.Mat Jais, 1997. Effects of Channa striatus Extracts on Morphine Analgesic. Presented at the 12th Scientific Meeting of the Malaysian Society of Pharmacology and Physiology, Kuala Lumpur, pp 1

3. Dambisya, YM, YM.; T.L.Lee; V.Sathivulu and AM.Mat Jais, 1999. Influence of Temperature, pH and naloxone on the Antinociceptive Activity of Channa striatus (Haruan) Extracts in Mice. J. Ethnopharmacol., vol. 66, pp 181 – 186

4. Che Ku Dahlan-Daud, A Zuraini, MA Abdah, A Aishah and AM Mat Jais, 2010. Proximate and Mineral Compositions of Haruan Channa striatus An Indigenous Malaysian Freshwater Fish. BLACPMA (in press)

5. Kumar, V., 1995. Haplotopic Differences and Genetic Variability Revealed by Mitochondria DN in Haruan Channa striatus Bloch. A Thesis Submitted to Faculty of Fisheries and Marine Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Malaysia for the Degree of Bachelor of Sciences (Fisheries)

6. Mat Jais; AM., R. McCulloch and K.D. Croft, 1994. Fatty Acids and Amino Acids in Haruan as a Potential Role in wound healing. Gen. Pharmacol., vol. 25(5), pp 947 – 950

7. Mat Jais, AM.; Y.M.Dambisya and T.L.Lee, 1997. Antinociceptive activity of Channa striatus Extracts in Mice. J. Ethnopharmacology, vol. 57, pp 125 – 130

8. Mat Jais, AM.; R.Fung; E.Bosi; C.Platell; R.McCauley and K.Croft, 1998. Preliminary Evidence on the Potential of Haruan. Malaysian Applied Biol., vol. 27, pp 50 – 51

9. Mat Jais, AM 2007. Molecular Size of the Bio-active Components from Haruan Channa striatus Extract. Journal of Applied Sciences, 7(15), pp 2198 – 2199

10. Mohsin, AK and MA Ambak, 1983. Freshwater Fishes in Peninsular Malaysia, pp 152 – 161. University Putra Malaysia (Publ) Serdang, Malaysia

11. Rahim, MHA., A.Rozila and AM Mat Jais, 2009. The Physical-Chemical and Morphological Study of Haruan Channa striatus in Peninsular Malaysia. Res J Biol Sci., vol 4(9), pp 994 – 1009

12. Somchit, MN., MH.Solihah, DA.Israf, A.Zuraini, AK.Arifah and AM.Mat Jais 2004. Effects of Three Local Malaysian Channa spp. Fish on Chronic Inflammation, Journal Oriental Pharmacy and Experimental Medicine. Journal Oriental Pharmacy an Experimental Medicine, vol 5(1), pp 91 – 94

13. Zakaria, ZA.; MN.Somchit; MR.Sulaiman and AM.Mat Jais, 2004a. Preliminary Investigation on the Antinociceptive Properties of Haruan Channa striatus Fillet Extracts with Various Solvent Systems. Pakistan Journal of Biological Sciences, vol 7(10), pp 1706 – 1710

14. Zakaria, ZA.; MN.Somchit; MR.Sulaiman, AM.Mat Jais and DA.Israf 2004b. Non-Opioid Antinociceptive Activity of Fresh Haruan (Channa striatus) Fillet Extract. Journal of Technology & Management, vol 2, pp 6 – 11

15. Zakaria, ZA.; AM.Mat Jais; MR.Sulaiman and MN.Somchit, 2004c. Effects of α-Amylase, Protease and Lipase on Haruan Channa striatus Mucus Extracts Antinociceptive Activity in Mice. Pakistan Journal of Biological Sciences, vol 7(12), pp 2202 – 2207

16. Zakaria, ZA.; MR.Sulaiman; MN.Somchit and AM.Mat Jais, 2005a. Evaluation of the Onset and Offset Time of Aqueous Supernatant of Haruan Channa straitus Fillet Antinociceptive Administered Subcutaneously in Mice. Pakistan Journal of Biological Sciences, vol 8(3), pp 466 – 469

17. Zakaria, ZA.; M.Safarul; R.Valsala; MR.Sulaiman; CA.Fatimah; MN.Somchit and AM.Mat Jais, 2005b. The Influences of Temperature and Naloxone on the Antinociceptive Activity of Corchorus olitorius L. in Mice. Naunyn-Schmiedeberg’s Arch Pharmacology, vol. 372, pp 55 – 62

18. Zakaria, ZA.; MR.Sulaiman; MN.Somchit; EC.Justin; I.Daud and AM.Mat Jais, 2005c. The Effects of L-Arginine, D-Arginine, L-NAME and Methylene Blue on Haruan Channa striatus – Induced Peripheral Antinociception in Mice. Journal of Pharmacy and Pharmaceutical Sciences, vol. 8, pp 199 – 206

19. Zuraini, A.; MN.Somchit; MH.Solihah; YM.Goh; AK.Arifah; MS.Zakaria; MN.Somchit; MA.Rajion; ZA.Zakaria and AM.Mat Jais, 2006. Fatty Acid and Amino Acid Composition of Three Local Malaysian Channa spp Fish. Food Chemistry, vol.97, pp 674 – 678


 Strength

Almost all natural product researchers locally as well as internationally, are working on plant species, and only few are on animal. In this case, Haruan research is exclusively belonging to Haruan Research Group (UPM), except Prof Saringat Baie, (School of Pharmacy, USM) who started to formulated cream for topical used in 1996/1997. This research had started in 1985 covering every aspect of drug discovery.

Based on genetic variability study revealed by mitochondria DNA (Kumar, 1995), Haruan C striatus is existing almost at the same time with Banjaran Titiwangsa some 600,000 years ago. Therefore, Haruan C striatus is indigenous to Malaysia, and this project is promoting our very own species into drug and bio-wealth.

All the formulations are steroid free, containing very minimal chemical additive, and they are purely natural products and fully organic. The products are 100 % Halal and has been registered in Triple Helix – Online Partnering at Biotech Corp, MOSTI, and samples of the products are in Figure 1 below.

Figure 1: Showing the Two Main OTC Products Using HTE as Therapeutic Agent, Medicated Cream and Capsules

Haruan that are produced in farming activities not only be used for OTC and pharmaceutical products, but also as food to help the country solving future food crisis. Marine fishes are getting depleted, birds and cattle are constantly attacked by diseases reduced confident by consumers, making freshwater species especially Haruan a good alternative. This is uponing-up opportunity to Malaysian involving in high-impact project of Haruan Agro-industry.

Supplies of high quality lives Haruan C striatus through farming is very consistent compared to many other natural product plant species that require years to be matured. Few of our herbs are not being certified which hindering exports, and to be used as raw materials in pharmaceutical products. Haruan only need 4 months reaching a 250 g size to be harvest and ready for production. This is a huge advantage and strength of this project for commercialization, and every Haruan produced is certified to qualified as raw materials for a GMP facilities and pharmaceutical products.


 Achievement

A Utility Innovation patent (UI 9088631 – process for preparation of fish Haruan Channa striatus extract having pharmaceutical function) has been awarded in 2009. Few more patent with commercial value will be filed soon.

There are three different forms of this in-water proprietary HTE (liquid or lHTE; solid or sHTE and nano-particles or nHTE) to be used seperately as therapeutic agent for specific products formulation. The lHTE is used in making tonic, the sHTE is for personal cares especially medicated cream, and nHTE is for tablet and capsule production.

DNA sequence of Haruan C striatus from three localities in Peninsular Malaysia had been registered at GenBank (GenBank Accession: JF 926031 – JF 826036) the nucleotide sequences. This truly an exciting achievement for many of our herbs are not being having much genetic background. With this registration, tt is becoming more evident and documentation for the commercialization of the products for global distribution.

The farming activities is now started at a small scale at Ulu Langgat, Selangor,, before going into commercial scale. The project is funded by a Techno Fund MARDI and under the management of Floa AQTech Sdn Bhd.

Prototypes of each selected products namely tonic, medicated cream and soft-gel capsule are to be outsourced at a GMP facilities where early negotiation and discussion has been made. These prototypes are for commercialization of OTC products, as well as clinical batches for clinical trials.


 

Facilities

Most of the R & D are carried out at Research Laboratories at Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, UPM. However, due to constrain of fund, some of the works were done in collaboration at other facilities with UPM (Faculty of Science and Food Technology, Faculty of Biotechnology and Molecular Sciences, and Faculty of Veterinary Medicine) including other facilities esoecially Institute of Biosciences. The project were also carried out in collaboration at other local universities/agencies (UM, UKM, UiTM, UMSabah, UNISEL, FRIM and Nuclear Angecy Malaysia) and foreign universities (Chulalongkorn, NUH Singapore, University of Southampton, UK, UWA Western Australia and Academia Sinica Beijing).

At the Faculty, the project is given an office room on level 3, Blok Makmal for Haruan Research Group and constantly used for meeting, consultation and student presentation.