PANICHELLI PAOLA (IT)
VALENTINI GIANLUCA (IT)
MARTINI DOMENICO (IT)
PANICHELLI PAOLA (IT)
VALENTINI GIANLUCA (IT)
PENTLOW K.S. ET AL.: "Quantitative imaging of iodine-124 with PET", THE JOURNAL OF NUCLAR MEDICINE, vol. 37, 1996, pages 1557 - 1562, XP002541428
| CLAIMS
1. New molecule 5-Hydroxy-l-(4-hydroxy-3-[ 124 I]iodo-5-methoxyphenyl)-7- (4-hydroxy-3-methoxyphenyl)-l,4,6-heptatrien-3-one having the following structural formula:
• conjugated therapeutic properties of curcumin and Iodine- 124 [ 124 I] through the Auger effect;
• possibility to define in a quantitative manner the distribution of the tracer at the level of the amyloid plaques in order to be able to devise personalized treatments;
• possibility to follow over time, up to 4 days, the variations of the tracer inside the amyloid plaques;
• better washout compared to analogous tracers; 2. Use of the new molecule 5-Hydroxy-l-(4-hydroxy-3-[ 124 I]iodo-5- methoxyphenyl)-7-(4-hydroxy-3-methoxyphenyl)- 1 ,4,6-heptatrien-3-one as claimed in claim 1 for:
• therapeutic treatment of Alzheimer's disease;
• PET diagnostic imaging; • pre- and post-therapy dosimetry investigations.
3. New molecule 5-Hydroxy-l-(4-hydroxy-3-[ 124 I]iodo-5-methoxyphenyl)-7- (4-hydroxy-3-methoxyphenyl)-l,4,6-heptatrien-3-one as claimed in claim 1 obtainable through three different processes of synthesis all of the same importance, the first process of which involves the adding, in an inert atmosphere, of 100 μl of HCl 1 N and 185 MBq OfNa 124 I in NaOH 0.02 N to a solution of 5-Hydroxy-7-(4-hydroxy-3-methoxyphenyl)-l-(4-hydroxy-3- methoxy-5-tributylstannylphenyl)-l,4,6-heptatrien-3-one in 100 μl of ethanol. 100 μl of H 2 O 2 (3% w/v) are added and the mixture is left to sit for 10 minutes at ambient temperature. After adding 300 μl of a solution saturated with NaHSO 3 , ethyl acetate is used to extract the product labelled with Iodine-124[ 124 I]. The ethyl acetate is evaporated under nitrogen flow and it is reconstituted in NaOH 0.5 N and a phosphate buffer with a pH of 7.4 is added. 4. New molecule 5-Hydroxy-l-(4-hydroxy-3-[ 124 I]iodo-5-methoxyphenyl)-7- (4-hydroxy-3-methoxyphenyl)-l,4,6-heptatrien-3-one as claimed in claim 1 obtainable with the second synthesis process which envisages the adding in sterile conditions and in a controlled atmosphere of 20 μl of an iodogen solution to CHCl 3 (1 mg of iodogen to 1 ml of CHCl 3 ), the evaporation of the solvent under nitrogen flow, the adding of 100 μg of 5-Hydroxy-l,7-bis(4- hydroxy-3-methoxyphenyl)-l,4,6-heptatrien-3-one (curcumin) to 10 μl of NaOH 0.5 N and 185 MBq of Na 124 I to NaOH 0.02 N. CHCl 3 is used to extract the product labelled with Iodine- 124 [ 124 I], the chloroform is made to evaporate under nitrogen flow and it is reconstituted in NaOH 0.5 N and a phosphate buffer with a pH of pH 7.4 is added. 5. New molecule 5-Hydroxy-l-(4-hydroxy-3-[ 124 I]iodo-5-methoxyphenyl)-7- (4-hydroxy-3-methoxyphenyl)-l,4,6-heptatrien-3-one as claimed in claim 1 obtainable with a third synthesis process by means of which, in inert atmosphere, 185 MBq OfNa 124 I in NaOH 0.02 N and 1.0 mg di choloramine- T are added to 100 μl of a 5.2xlO '2 M solution, in THFiH 2 O 50%, of precursor 5-Hydroxy-7-(4-hydroxy-3-methoxyphenyl) 1 - (4 -hydroxy-3- methoxy-5 -potassiumtrifluoroboratephenyl)- 1 ,4,6-heptatrien-3 -one, after which the mixture is left to sit for 10 minutes at ambient temperature. After adding 100 μl of the 1.OxIO "4 M solution Of NaHSO 3 , the product labelled with Iodine- 124[ 124 I] is isolated transferring the solution by means of a silica Sep-Pak cartridge. The product labelled with Iodine- 124[ 124 I] is eluted with a solution of petroleum ether: ethyl acetate (50: 1). The solvent is made to evaporate under nitrogen flow and it is reconstituted in NaOH 0.5 N and a phosphate buffer with a pH of 7.4 is added. 6. New molecule 5-Hydroxy-l-(4-hydroxy-3-[ 124 I]iodo-5-methoxyphenyl)-7- (4-hydroxy-3-methoxyphenyl)-l,4,6-heptatrien-3-one as claimed in claims 1, 2, 3, 4 and 5, characterized by the fact that it can be administered by general oral route in the form of tablets, soft or hard gelatin oil or operculated capsules, sugar-coated pills, dispersing powders, suspensions or emulsions or by topical or transdermal route in the appropriate forms and in vehicles or devices suited for the administration of the active principle at the site of interest.
7. New molecule 5-Hydroxy-l-(4-hydroxy-3-[ 124 I]iodo-5-methoxyphenyl)-7- (4-hydroxy-3-methoxyphenyl)-l,4,6-heptatrien-3-one as claimed in claims 1, 2, 3, 4 and 5, characterized by the fact that it can be administered by general parenteral route in the form of aqueous or oily solutions, or suspended in appropriate dispersing agents, even in the form of lyophilized products, to be dispersed at the time of the administration. |
Title
New molecule 5-Hydroxy-l-(4-hydroxy-3-[ 124 I]iodo-5-methoxyphenyl)-7- (4-hydroxy-3-methoxyphenyl)-l,4,6-heptatrien-3-one for PET investigations and radiotherapy.
Field of invention
The present invention concerns a new molecule, 5 -Hydroxy- l-(4-hydroxy-3- [ 124 I]iodo-5-methoxyphenyl)-7-(4-hydroxy-3-methoxyphenyl)-l,4,6- heptatrien-3-one, ideal for use in PET diagnostics and radiotherapy. Background
PET (Positron Emission Tomography) is a diagnostic method through which the metabolism of the various organs under examination may be studied, in order to obtain early diagnostic parameters for various diseases. Over recent years, the use of this method has expanded considerably in the USA and PET diagnostic centres are currently increasing all over Europe, mainly in the oncology sector. In this sector, PET diagnostics introduces two clinical parameters, early diagnosis and optimization of treatment, which change the life expectations of patients and improve the management of their illness. Other fields of application are acquiring an increasingly important role, such as neurology, cardiology and rheumatology.
In an era in which the average life span is continuously increasing, PET is destined to play a role of major importance in the study of age-related diseases such as Parkinson's and Alzheimer's disease, as it has had in the study of molecules by means of which it is possible to make early diagnoses of acute cardiac events which are, nowadays, one of the main causes of death. PET is carried out by injecting a radiopharmaceutical into the patient and following the distribution of the radiopharmaceutical inside the human body with special machines called PET (Positron Emission Tomography) scanners.
The radiopharmaceuticals are composed of two essential parts: a radioisotope (which emits beta rays) and a molecule which binds with the radioisotope, constructing the metabolic substrate of the PET investigation. The radioisotopes are produced by an instrument called a Cyclotron and they are bound to the molecule to be studied by specific methods of chemical synthesis.
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Journal of Nuclear Medicine and Molecular Imaging 23 November 2004. Another rapidly expanding sector in PET diagnostics is that of the development of radiotherapy treatment plans through which the therapeutic effect can be optimized. In this sense, there are various phases of development in the research carried out in this sector which spans from the demonstrated usefulness of FDG to the recent experiences with the isotope copper-64, in particular the molecule 64 Cu-ATSM through which it is possible to obtain a map of the distribution of oxygen inside the tumour mass. 53. Molecular mechanism of copper uptake and distribution; Current
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PET provides the possibility, with the use of specific tracers, to implement treatment plans that are more precise and in quantitative measures, both for metabolic therapy and for radiotherapy.
The most interesting tracer for metabolic therapy is Iodine- 124[ 124 I] which makes it possible to quantify thyroid lesions. It is currently used in conventional nuclear medicine, but with rather unsatisfactory and sketchy results. Iodine- 124 [ 124 I] is an unstable isotope that does not exist in nature. It has a half-life of 4/18 days and is produced through a nuclear reaction: 124 TeO 2 enriched by 99.8 % with 124 TeO 2 using proton energy in a range of 14-10 MeV. This nuclear reaction brings the 124 I to a very high degree of purity compared to 125 I and 126 I, the purity levels of which are lower. In fact, the radionuclide Iodine- 124 [ 124 I] decays through the decay of β + and electronic capture, including in its emission spectrum both the Auger effect - an emission of great importance for therapy - and the gamma radiation used in tomographic imaging.
Iodine- 124 [ 124 I] is therefore an ideal isotope when used as a radiotracer in nuclear medicine for positron emission tomography (PET).
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Iodine- 124 [ 124 I] in the form Of Na 124 I is used for PET analysis and its long half-life (4/18 days) enables both the development of multiple radiochemical syntheses and the detection of slow biochemical processes that could not be
1 1 1 δ detected using tracers with a short half-life such as C and F. Iodine- 124[ 124 I] in the form Of Na 124 I is used in radio immunodiagnosis and radio immunotherapy as a dosimetry indicator to verify the status of the therapy with 131 I, used in the treatment of thyrotoxicosis and tumours of the thyroid.
With Iodine-131( 131 I), however, a PET investigation is not possible because its energy is too high and it cannot be detected by the instrument. For this
reason it is used during radio immunotherapy with Iodine- 124 [ 124 I] as this enables the progress of the therapy to be followed through PET imaging. Iodine- 124 [ 124 I] was not considered to be suitable for PET imaging studies because it is a low emitter of positrons. In 1996, Pentlow demonstrated that despite the fact that Iodine- 124[ 124 I] was a low emitter of positrons, it was nonetheless suitable for detecting tumours surrounded by relatively low background activity, such as thyroid diseases. In fact, the PET analysis with Iodine- 124[ 124 I] is the ideal technique for studying the kinetics of iodine in thyroid cancer therapy. There are essentially two advantages to be gained from the use of Iodine- 124[ 124 I] in nuclear medicine compared to the diagnostic application currently in use, i.e. Iodine-123[ 123 I] with the SPECT technique:
• possibility to define in a quantitative manner the distribution of the tracer at the basal nuclei. This is only possible in a semi-quantitative manner with Iodine-123[ 123 I] in SPECT;
• possibility to follow over time, up to 4 days, the variations of the tracer inside the basal nuclei. This also permits the conducting of pharmacological tests and the assessment of their results.
An important application, already object of the Italian national patent application no. MC2007A000196 of 10/10/2007 (inventors: Martini
Domenico, Panichelli Paola, Valentini Gianluca), is iodine 124βCIT Iodine-
124[2β-carbomethoxy-3β-(4-iodophenyl)-tropane] which enables the effecting of presynaptic diagnostics of the activities of the corpus striatum in a quantitative manner, with the possibility to run pharmacological tests at a lower cost compared to iodine- 123 βCIT iodinel23βCIT Iodine- 124 [2 β- carbomethoxy-3β-(4-iodophenyl)-tropane] and with a marked increase in quality.
Alzheimer's Disease (AD) is a disease associated with progressive cognitive and mental degeneration. The anatomopathological detail that distinguishes it
is the presence of cerebral deposits of amyloid-beta peptide plaques with intracellular filaments which contain the hyperphosphorylated protein tau. The depositing of these plaques in the cerebral substance can take place prior to the emergence of the symptoms typical of the disease and is a diagnostic target on which studies have been focusing for many years now.
The radiopharmaceuticals that show up deposits of amyloid plaques in the cerebral tissue represent the more advanced diagnostic phase for defining the early stages of the disease. In this sense various radioligands have been identified, the first used in studies on humans, in 2002, was 18 FFDDNP by means of which amyloid plaques could be viewed in vivo. Subsequently, in 2004, PIB, known as the Pittsburgh compound, was used experimentally. Another tracer used is π Csb-13, a derivative of stilbene which has proved to possess a moderate lipophilicity and a rapid brain washout. Experiments were carried out in 2007 using tracers bound to Iodine- 125 [ 125 I], such as [ 125 I]IMPY and [ 125 I]TZDM, with a benzothiazole structure.
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101. Brian J. Bacskai, Gregory A. Hickey, Jesse Skoch, Stephen T. Kajdasz, Yanming Wang, Guo-feng Huang, Chester A. Mathis, William E. Klunk,
Bradley T. Hyman Source: Four-Dimensional Multiphoton Imaging of Brain Entry, Amyloid Binding, and Clearance of an Amyloid-β Ligand in Transgenic Mice. National Academy of Sciences, 2003; 21 : 12462-12467.
102. Yanming Wang, Chester A. Mathis, Guo-Feng Huang, Manik L. Debnath, Daniel P. Holt,Li Shao, and William E. Klunk. Effects of
Lipophilicity on the Affinity and Nonspecific. Binding of Iodinated Benzothiazole Derivatives. Journal of Molecular Neuroscience, 2003; 20:255-260.
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111. B. Neumaier, S. Deisenhofer, D. Fϋrst, C. A. F. von Arnim, S. Thees, A. K. Buck, G. Glatting, G. B. Landwehrmeyer, B. J. Krause, H. D. Mϋller, C. Sommer, S. N. Reske, F. M. Mottaghy. Radiosynthesis and evaluation of [HCJBTA-land [HC] 3'-Me-BTA-I as potential radiotracersfor in vivo imaging of β-amyloid plaques. Nuklearmedizin, 2007; 46: 271-280.
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An important application, already object of the Italian national patent application No. MC2008A000089 of 23/05/2008, (inventors: Martini Domenico, Panichelli Paola, Valentini Gianluca), is Iodine- 124 [2-(3'-iodo-4' - methylaminophenyl)-6-hydroxy-benzothialzole] by means of which PET diagnostic imaging can be carried out and it presents a better washout compared to similar substances labelled with Carbon- 11 [ 11 C] or with Fluoro- 18[ 18 F]. Curcumin, a phenolic substance, is the main biologically active component in Turmeric. The chemical formula of curcumin is C21H20O6, it is known as diferuloylmethane and its structural formula is as follows:
In contrast with many other antioxidants, curcuminoids have the capacity both to prevent the formation of free radicals and to neutralize already existing free radicals, and they are considered to be efficacious bioprotectors due to this dual action.
Independent studies on the anti-cancer properties of Turmeric suggest that Curcumin is potentially useful for the treatment of various types of cancer: colon, breast, prostate, lung and skin cancer among others. Numerous mechanisms of action for the anti-tumour activity of Curcumin have been described. The inhibition of the proliferation of tumour cells, the induction of apoptosis (a mode of cell death), the inhibition of the transformation of cells from normal to tumoral, the inhibition of invasion and metastasis and the suppression of inflammation have been connected with the anti-tumour activity of Curcumin. The inhibition of COX-2 and 5 -LOX, adhesion molecules, inflammatory cytokines, growth factor receptors, vascular endothelial growth factor (VEGF) and Curcumin transcription factors are closely connected with its anti-tumour activity.
Studies in vitro and in vivo have proved that curcumin has the capacity to prevent carcinogenesis at three different levels: tumour promotion, angiogenesis and tumour growth (Maheshwari et al., 2006).
Recent studies have demonstrated that curcumin has a dose-dependent, chemopreventive effect in various animal models of oral carcinogenesis (colon, duodenum, stomach, oesophagus), and the low incidence of intestinal tumours in Indians has been attributed to the use of Turmeric in food (Mohandas, Desai, 1999).
The molecular base of the anti-carcinogenic and chemopreventive activity is attributed to its effect on different targets, including various transcription factors, growth regulators, adhesion molecules, apoptotic genes, angiogenesis regulators and intracellular signal molecules (Aggarwal et al., 2003). In the rat, curcumin administered by oral route inhibited the expression induced by carcinogens of the c-Ha-ras and c-proto-oncogenes in skin tumours (Pornngarm et al., 2001). Conjugated compounds were assayed for their apoptosis-inducing properties on tumour cells, and their activity is well correlated with the generation of ROS by the cells, while the levels of GSH remained the same (Mishra et al., 2005). The study also indicated a downregulation of Bcl-2 and the participation of caspasi-3 in the apoptotic death of tumour cells. Antiproliferative and pro-apoptotic activity has also been observed in the cells of a human ovarian carcinoma (Shi M. et al., 2006). Curcumin also possesses anti-angiogenic properties, reducing the expression of pro-angiogenic genes such as VEGF (Vascular endothelial growth factor) and some metalloproteases (Sarvjeet and Ashok., 2006). Arthritis is an inflammatory disease. All the current pharmaceuticals approved for arthritis are anti-inflammatory. Anti-TNF (tumour necrosis factor) therapy has recently been approved for this disease. It has been demonstrated that Curcumin has the capacity to stop the production of TNFs and block the action of TNFs. Topically applied, curcumin has been proved to exert an action against arthritis.
Crohn's disease is an inflammatory disease. All the current pharmaceuticals approved for this disease are anti-inflammatory. Anti-TNF therapy has been approved for this disease. Curcumin has been demonstrated to block both the production and the action of TNF. Curcumin administered by oral route has proved to exert an action against Crohn's disease.
Numerous experimental data suggest that Curcumin can accelerate the healing of wounds. This induced Johnson & Johnson, for example, to market and sell Band Aid sticking plasters containing curcumin in India. Psoriasis is another inflammatory disease. Relevant elements of proof, both in animals and in humans, indicate that Curcumin is very efficacious against psoriasis when applied topically to the skin.
The findings emerging from studies with radio-labelled curcumin were significant absorption followed by rapid transformation into a metabolite. Curcuma longa, and its active ingredient, curcumin, exert an interesting antiinflammatory activity both on the lipoxygenase system and on the cyclooxygenase system. It also exerts an antimutagenic, antioxidant and chemopreventive activity on several models in vitro and it is currently one of the most important chemopreventive agents under study for the prevention of neoplasms. Anti-HIV activity has been documented in vitro with an action on the LTR, on various proteases, on cytokines such as TNF-alfa and on integrase. Research on humans has produced conflicting results.
Various studies have discovered that Curcumin can help the immune system "clean" the brain of amyloid-beta, the main constituent of the plaques found in Alzheimer's disease. Since Alzheimer's disease is partly caused by inflammation induced by amyloid, Curcumin has proved to be efficacious against Alzheimer's disease. Clinical trials are currently in course at UCLA (University of California, Los Angeles) on the use of Curcumin for Alzheimer's disease. A study carried out by Italian and US researchers suggests that curcumin could increase the protection and antioxidant properties of an enzyme that protects the brain.
The scientific team identified an innovative compound that acts as an inducer of heme oxygenase- 1 (HO-I), a gene of the central nervous system that codes
for a protein with antioxidant properties. The function of this protein is to protect the neurons of the brain exposed to oxidation processes. Curcumin and the yellow pigment of curry are thought to have neuroprotective properties and to increase, at the cytoplasmatic and nuclear level, the expression of the protein heme oxygenase- 1. Although the results of this study are very encouraging, several studies in vitro and in vivo are required in order to ascertain whether curry effectively has preventive properties against acute neurodegenerative disorders. Important studies also exist regarding the potential use of curcumin in the prevention of neurodegenerative diseases, particularly Alzheimer's disease. In fact, as already mentioned earlier, both oxidative damage and inflammatory processes are particularly high in the brains of patients affected by Alzheimer's disease, suggesting the possible usefulness of a long-term therapy with a compound possessing powerful antioxidant and anti- inflammatory properties.
In particular, a number of studies were carried out on heme oxygenase- 1 (HO- 1), an inducible and redox-sensitive protein that exerts a cytoprotective action against oxidative stress. This protein degrades heme to the vasoactive molecule carbon monoxide and the antioxidant biliverdin. Curcumin induces the expression of heme oxygenase- 1 in vascular endothelial cells (Motterlini et al., 2000). This data is borne out by a study demonstrating how the phenyl- ethyl ester in Caffeic acid phenethyl ester (CAPE), another plant-derived phenolic derivative, markedly increased heme oxygenase activity and HO-I protein in astrocytes (Scapagnini et al., 2002). The effect, not necessarily dependent on a glutatione-mediated mechanism, seemed to be related to the peculiar chemical structures of curcumin and CAPE, because analogous antioxidants containing only portions of these two molecules were totally ineffective. Similar results were obtained with Curcumin-95, a mixture of curcuminoids commonly used as a dietary supplement. The treatment of
astrocytes with curcumin regulated the expression of the HO-I protein both at a cytoplasmatic and at a nuclear level and an increase in quinone reductase and glutathione S transferases was observed in the astrocytes exposed to 5-15 micromolar curcumin (Scapagnini et al., 2006). Additionally, the effects of this molecule were also assayed on hippocampal neurons in culture. High expression of HO-I was observed after 6 hours of incubation with 5-25 micromolar curcumin.
Recent studies have connected the heme oxygenase pathway with the protective anti-degenerative mechanisms that develop in Alzheimer's disease. It has, in fact, been demonstrated by some Authors (Butterfield et al., 2002) that the expression of HO is closely connected with that of the amyloid precursor protein (APP). The induction of HO, which takes place at the same time as that of other stress-related proteins in various physiopathological conditions, and the consequent production of carbon monoxide and biliverdin, could prove to be a protective system against oxidative brain damage. This discovery is also backed up by recent epidemiological studies that demonstrate the extremely low incidence, in comparison with other parts of the world, of Alzheimer's and Parkinson's disease in the Indian population, where curcumin is used daily in the diet. Some Authors, however, point out the need for clinical trials with curcumin in patients affected by Alzheimer's disease as well as the opportunity to assess the effects of this molecule on the biomarkers of the disease.
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Disclosure of Invention
The new molecule 5-Hydroxy-l-(4-hydroxy-3-[ 124 I]iodo-5-methoxyphenyl)- 7-(4-hydroxy-3-methoxyphenyl)-l,4,6-heptatrien-3-one having the formula:
• conjugated therapeutic properties of curcumin and Iodine- 124 [ 124 I] through the Auger effect; • possibility to define in a quantitative manner the distribution of the tracer at the level of the amyloid plaques in order to be able to devise personalized treatments;
• possibility to follow over time, up to 4 days, the variations of the tracer inside the amyloid plaques; • better washout compared to analogous tracers; to be used:
• for therapeutic treatment of Alzheimer's disease;
• for PET diagnostic imaging;
• for pre- and post-therapy dosimetry investigations. Detailed description
The new molecule 5-Hydroxy-l-(4-hydroxy-3-[ 124 I]iodo-5-methoxyphenyl)- 7-(4-hydroxy-3-methoxyphenyl)-l,4,6-heptatrien-3-one can be obtained through three different processes of chemical synthesis, all of the same importance, and all starting from the radioisotope Iodine- 124 [ 124 I]. For the production of the radioisotope Iodine- 124[ 124 I], a 18 MeV IBA cyclotron is used, with a solid target (COSTIS) dedicated to the formation of Cu-64 and Iodine- 124[ 124 I].
The method consists of bombardment with protons for approx. eight hours at a current of 18 μA, on enriched Tellurium oxide 124 TeO 2 , mass of tellurium oxide 250 mg, on a platinum disc (target substrate).
The yield of the bombardment is approx. 3.7 GBq of Iodine- 124 [ 124 I], while the energy of the beam has a maximum cross section of 14 Mev. Additionally, during irradiation of the solid targets, it is important for the proton beam to be perfectly centred; to do this, it is essential to know its shape. This is detected by a special autoradiography scanner (Cyclone) by means of which the image of a previously irradiated aluminium disc is recorded, on a phosphorus film. The substrate used for the Target is a Platinum disc with a diameter of 24 mm and a circular cavity of 12 mm, which guarantees good conductivity and resistance to corrosion, and a mixture of isotopically enriched tellurium oxide 124 TeO (95%) and aluminium oxide Al 2 O 3 (5%) weighing approx. 250 mg is used, so that the aluminium acts as a binding agent for the crystal matrix. The mixture is smelted at 753 0 C and re-solidified in a quartz furnace for approx. 2 hours . Once the target is ready - and it must be as stable as possible against the high bombarding current in order to minimize any losses Of TeO 2 when the vapour pressure reaches high values - bombardment commences. The separation of Iodine- 124[ 124 I] from the matrix of the target disc ( 124 Teθ 2 /alumina) takes place by means of a thermodistillation process carried out using the TERIMO - "Automatic 1247123 I iodine isotope synthesis module for PET scanning System control of 1241 radioactive iodine isotope synthesis".
The control system is based on a PLC, a temperature controller and an air flow regulator, and a Scada System used for the control and acquisition of data. The Iodine- 124[ 124 I] recovery time is approx. an hour and the mix is smelted at 753°C releasing 124 I 2 in the form of gas; the 124 I 2 is bubbled through a hyperpure solution of NaOH 0.02 N; the 124 I 2 is entrapped in the NaOH solution in the form of [ 124 I]NaI (sodium iodide at 95%), sodium iodate NaIO 3 and periodate NaIO 4 (5%).
The entire chemical process takes place inside the synthesis module placed in a glove box in order to ensure the product's sterility in accordance with GMP guidelines which guarantee a product, the main features of which are quality and efficacy. At the end of the process, we obtain [ 124 I]NaI which may be administered as a radiotracer on its own or used as a radiolabel for the synthesis of new radiopharmaceuticals.
The radionuclidic purity of Iodine- 124[ 124 I] is obtained using a Germanium Gamma ray Spectrometer in order to detect the presence of Iodine-125[ 125 I], Iodine-126[ 126 I], Iodine- 130[ 130 I], Iodine-131[ 131 I]; these impurities must be lower than 0.1%.
Observing the peaks of energy, the purity of the radionuclide (excluding the presence of Iodine- 123 [ I] which is, in any case, at a bare minimum) must be greater than 99.5%. Once the Iodine- 124 [ 124 I] radioisotope has been obtained, the first chemical synthesis process consists of adding 100 μl of HCl 1 N and 185 MBq of Na 124 I in NaOH 0.02 N in an inert atmosphere to a solution containing 100 μg of precursor 5-Hydroxy-7-(4-hydroxy-3-methoxyphenyl)-l-(4-hydroxy-3- methoxy-5-tributylstannylphenyl)-l,4,6-heptatrien-3-one in 100 μl of ethanol. 100 μl di H 2 O 2 (3% w/v) are then added after which the mixture is left to sit for 10 minutes at ambient temperature. After adding 300 μl of a solution saturated with NaHSO 3 , ethyl acetate is used to extract the product labelled with Iodine- 124 [ 124 I]. The ethyl acetate is evaporated under nitrogen flow and it is reconstituted in NaOH 0.5 N and a phosphate buffer with a pH of 7.4 is added.
The second synthesis process involves the adding - in sterile conditions and in a controlled atmosphere - of 20 μl of an iodogen solution to CHCI 3 (1 mg of iodogen to 1 ml of CHCl 3 ), evaporation of the solvent under nitrogen flow, the adding of 100 μg of 5-Hydroxy-l,7-bis(4-hydroxy-3-methoxyphenyl)-
l,4,6-heptatrien-3-one (curcumin) to 10 μl of NaOH 0.5 N and 185 MBq of Na 124 I to NaOH 0.02 N. CHCl 3 is used to extract the product labelled with Iodine- 124 [ 124 I], the chloroform is made to evaporate under nitrogen flow and it is reconstituted in NaOH 0.5 N and a phosphate buffer with a pH of pH 7.4 is added.
The third synthesis process involves a treatment in inert atmosphere, with 100 μl in a 5.2x10 '2 M solution, in THFiH 2 O 50%, of precursor 5-Hydroxy-7-(4- hydroxy-3-methoxyphenyl) 1 - (4 -hydroxy-3-methoxy-5- potassiumtrifluoroboratephenyl)-l,4,6-heptatrien-3-one, the formula of which is:
185 MBq Of Na 124 I in NaOH 0.02 N and 1.0 mg of choloramine-T are added and the mixture is left to sit for 10 minutes at ambient temperature. After adding 100 μl of a l .OxlO "4 M solution Of NaHSO 3 , the product labelled with Iodine-124[ 124 I] is isolated transferring the solution by means of a silica Sep- Pak cartridge. The product labelled with Iodine- 124 [ 124 I] is eluted with a solution of petroleum ether: ethyl acetate (50:1). The solvent is made to evaporate under nitrogen flow and it is reconstituted in NaOH 0.5 N and a phosphate buffer with a pH of 7.4 is added. The new molecule, produced according to the synthesis processes described above, is prepared by dissolving the active substance in salt solution; filtration of the solution with a 0.22 μm filter (with sterilizing effect, followed by the immediate transfer of the filtrate in a sterile block), distribution into the bottle-shaped containers.
Since the active substance for each container is in quantities that are too small and the corresponding volume of solution would be difficult to dose accurately, in addition to the fact that at the end of the process the vial or bottle would appear almost empty, it is necessary to add an atoxic excipient, free from any kind of pharmacological activity that gives the freeze-dried product an attractive appearance: 20 mg of mannitol.
The product to be treated is frozen at a temperature of -40 0 C inside stainless steel containers (that meet health and hygiene standards) which are, in turn, placed inside the lyostat in which the pressure is reduced to a value such that the water present in the previously frozen product may sublimate under vacuum by heating to a temperature of 30° C, leaving the product almost completely dried.
In this way a solid, porous, crumbly, hygroscopic mass is obtained that is very soluble in solvent, and that occupies the same volume of the initial frozen, or freeze-dried, mass.
The new molecule 5-Hydroxy-l-(4-hydroxy-3-[ 124 I]iodo-5-methoxyphenyl)- 7-(4-hydroxy-3-methoxyphenyl)-l,4,6-heptatrien-3-one may be administered by general oral route in the form of tablets, soft or hard gelatin oil or operculated capsules, sugar-coated pills, dispersing powders, suspensions or emulsions or by topical or transdermal route in the appropriate forms and in vehicles or devices suited for the administration of the active principle at the site of interest; it can also be administered through general parenteral route in the form of aqueous or oily solutions, or suspended in appropriate dispersing agents, even in the form of lyophilized products, to be dispersed at the time of administration.
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