Dioraphte Foundation will fund an official PK trial, performed by XENDO (www.xendo.nl). The extract for this trial will be produced according to GMP guidelines by Conforma/University of Ghent. Luc Verelst (www.reliablecancertherapies.com) is funding the production.
Crude Artemisia extract produced from 2% yielding plants will cost $ 36 / kg, with a content of 23% artemisinin.
To produce for example an ACT consisting of 500 mg artemisinin* and 400 mg naphtoquine would cost:
artemisinin: $ 0,08
groundnut oil: $ 0,02
naphtoquine: $ 0,03
Packaging and quality control: ?
* The ACT Artemisinine-Naphtoquine contains 1000 mg artemisinin. Because bio-availability of artemisinin is increased we calculate with 500 mg artemisinin
A pilot-proof of concept trial was performed with 8 healthy volunteers (6 male, 2 female) in a cross-over design. The volunteers took artemisia-extract with oil containing 142 mg artemisinin as a single dose. Plasma-samples were collected at 0, 0.5,1,1.5,2,3,4,8,12 and 24 hours. Artemisinin was measured in the samples using HPLC-MS-MS
The same volunteers took 500 mg pure artemisinin as a single dose a few weeks later and plasma samples were collected again.
(Extract AUC/concentration) / (Artemisinin AUC/concentration) was for 7 of the volunteers:
3.8 , 3.7 , 2.1 , 2.1 , 1.8 , 1.8 , 1.1
The average increase in bio-availability of artemisinin was 136%
One volunteer had lower artemisinin plasma concentrations after taking the extract than after taking pure artemisinin ( - 66%). It turned out that this volunteer had been drinking heavily the night before the trial with the extract, not before the trial with pure artemisinin.
It seems that bio-availability of artemisinin can be increased a few times by means of using a simple artemisia extract mixed with oil as the source of artemisinin.
This pilot-trial was performed with a simple mixture of artemisia-extract with oil and under sub-optimal standardised conditions.
Further trials should be performed after homogenisation of the product.
The oil used by prof. Yao-De Wan was refined groundnut oil (1)
By HPLC-UV analysis (2) an artemisinin content of 5,3 mg/ml was established in the concentrated Artemisia extract of Scott Process Technology Ltd. That means that 18,9 ml Artemisia extract corresponds to 100 mg artemisinin. For 6 doses of 100 mg each, 113 ml of concentrated extract will be be used. To the 113 ml concentrated extract 25 ml of refined groundnut oil will be added. After evaporation of the ethanol in a rotary evaporator at 40 0C and a pressure of 80- 180 psi, the volume of the emulsion will be adjusted to 30 ml with refined groundnut oil (Acros organics, New Jersey, USA). The final emulsion/solution will contain 100 mg artemisinin/5ml. The emulsion/solution will be ready for preclinical studies after control of the artemisinin content by NMR.
Any advice on formulation is welcome
1: Wan YD, Zang QZ, Wang JS. Studies on the antimalarial action of gelatin capsule of Artemisia annua Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi.1992;10(4):290-4. Sichuan Institute of Chinese Materia Medica, Chongqing
2: The artemisinin concentration of the extract was determided using HPLC-UV by Jorge Ferreira and Barry Harter, USDA-ARS/AFSRC
The ratio of artemisinin/camphor in the Artemisia annua extract = 100 mg/22mg (Artemisinin determined by HPLC-UV, Camphor determined by GC-MS) The maximum exposure to camphor will be 1,1mg/kg bw/day for a period of three days. The EFSA advices that exposure to camphor does not exceed 2 mg/kg bw on a single day in any age group
European Food Safety Authority; Camphor in flavourings and other food ingredients with flavouring properties - Opinion of the Scientific Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Food on a request from the Commission Question number : EFSA-Q-2003-144
http://www.efsa.europa.eu/EFSA/efsa_locale-1178620753812_1211902029226.htm
Niemeijer Foundation and Farmacie Mondiaal (www.farmaciemondiaal.nl) donated each 6.000 euro to further investigate new methods for the purification of ethanolic primary extracts. The project is headed by prof. von Freyhold, University of Bremen. Results of these studies might lead to the emergence of factories in Europe that are able to process ethanolic primary extracts into ACT’s untill African countries have gained the expertise to do so themselves. In turn, this might enable African countries to produce concentrated ethanolic primary extracts in large quantities without first having to deal with the complex technical and marketing aspects of such a venture.
Transport of the concentrate from Africa to Europe would add roughly $0,5 to the price of 1kg pure artemisinin
Wednesday, 24th september 2008
Scott Process Technology (www.scottprotec.com) produced 40 liters of concentrated ethanolic Artemisia Annua extract. The concentrate was produced according to the extraction protocol developped by Dr. Al Fleming and Prof. Michaela von Freyhold for Medicines for Malaria Venture (http://www.mmv.org/IMG/pdf/3_ethanolic-extraction-december-2007.pdf )
40 kg of dried Artemisia Annua Silves leaves was extracted using 200 litres of pure ethanol. The Artemisia was provided by Prof. Pedro Melillo de Magalhães, Centro Pluridisciplinar de Pesquisas Químicas Biológicas e Agrícolas, Agrotechnology (CPQBA), University of Campinas, Brasil (http://www.icei.it/pdf/State_of_the_art_Artemisia_ICEI.pdf ). The ethanol was removed using a Thin Film Evaporator under vacuum.
Drs. Henk Goris is now analysing the extract for artemisinin as well as campher concentration.
Prof. Oliver Kayser is now analysing methods to produce a suitable and stable oil-based syrup of the extract. This syrup will be ingested by Dutch human volunteers in order to check the product for side-effects and blood plasma levels of artemisinin (pharmacokinetics).
You can read the full SPT-report here: (comming soon)
The Department of Farmaceutical Biology of the University of Groningen in association with the Hanze University Groningen recently received a 125.000 euro grant from Foundation Dioraphte to investigate a novel approach towards the production of Artemisinin Combination Therapy’s (ACT’s). The project will be headed by prof.dr. Oliver Kayser.
Information: prof.dr. O. Kayser, tel. 050-363 3299, e-mail:o.kayser@rug.nl
