It has been 3 a.m now. Last night I slept early coz I felt so tired after all day activities. I wake up from my dream wherein I met with my parents, yah, my mom and my daddy. Not too clear what the story was , it so fast to wake up. But, I feel so happy being able to see them even though just in a dream. I just realize that this early morning (2 May) is the same date when my daddy passed away 19 years ago. He died at the night after my birthday, and I think he want to give me a birthday wishing in this early morning, he3x….
My daddy was a person who had a straight and discipline character. He would undoubtedly hit me or my brothers when we made a mistake. Although I felt unguilty and my friend initiate me fighting first, he still punished me. I was not too close with him since we have in contrast character. I tend to be melancholies and I prefer to share with my mom rather than him. I thought he didn’t love me, because he never showed it. He never praised me when I got 1st rank in my school, he won’t give me a money to buy a book. I was really disappointed with him and jealous when I saw my friends having fun with their fathers.
But what we saw doesn’t always describe what the truth. I really felt what a huge he loved me when I got DENGUE fever and should be in a hospital patient. I saw what a panic he was more than my mom. I felt so peace when he hold me alike don’t want let me went away. Thereafter, I changed my perception and proven what a noble he was. He just wanted us to be an independent person, so we will be survived even in the critical situation.
My daddy was a hard smoker and had a complicated cardiovascular disease then it be worsted by his chronic ASTHMA. His physic was dropping triggered by asthma attack and it became worst when it modulated other cardiovascular system such as hypertension. Only in 6 hours after he dropped, he died at a night after my birthday. Ya…that was a destiny and nobody could avoid this. My big brother told me one last message he gave us that we had to be survive even not together anymore. I was crying so much and feel so sorry that I was thinking he never loved me.
Life must go on, and my goodness we have been survived even my mom being a single parent until now. She growths up us such as she never thinking herself. She dedicates the whole of her life to take care us. And step by step we can recover and being a compact family.
My carrier was getting better when I took a course in Master Degree. I chosen drug discovery and development as my field and specialized in SYNTHETIC MEDICINAL CHEMISTRY. Then I was inspired to study the drug discovery in ASTHMATIC disease regarding what my daddy was suffered.
Asthma is a common chronic inflammatory disease of the airways characterized by variable and recurring symptoms, airflow obstruction, and bronchospasm. Symptoms include wheezing, cough, chest tightness, and shortness of breath.
Certain key steps in the pathophysiology of asthma are depicted, emphasizing the role of the cysLTs C4, D4, and E4. The contributions of many cells (such as dendritic cells, mast cells, macrophages and epithelial cells) and mediators (such as proteases, eotaxin and endothelin) have been omitted. As shown, antigen challenge results in the accumulation and activation of TH2 lymphocytes, which secrete eosinophil-activating cytokines. Eosinophils in turn secrete cysLTs and other pro-inflammatory mediators. A variety of cell types also contribute to the generation of LXA4. CysLTs and LXA4 bind to target cells, including leukocytes, smooth-muscle cells, epithelial cells and endothelial cells. LXA4 can interact with its cognate receptor, ALX, to generate suppressive signals that are anti-asthmatic. In addition, LXA4 can compete with the binding of cysLTs at their cognate receptor, cysLT1, thereby blocking the pro-asthmatic actions of cysLTs. Inset: Metabolism of arachidonic acid (AA) to prostaglandins, cysLTs and LXA4. Other abbreviations: PLA2, phospholipase A2; COX, cyclooxygenase; LO, lipoxygenase.
One of classic asthmatic drug has been known is theophylline. Its mechanism action in asthmatic disease treatment was known by inhibiting Phosphodiesterase inhibitor (PDE4), one of enzymes in our bodies who catalyses the hydrolysis CAMP (Cyclic-Adenosine Mono Phosphate) become AMP (Adenosine Mono Phosphate). Bu inhibiting the hydrolysis pathway, the increasing of CAMPO will cause the smooth muscle in trachea or bronchus relax back.
Other study showed that theophylline can treat asthmatic disease by adenosine antagonism. The blocking of Adenosine Receptor with its natural ligand will prevent the smooth muscle constriction become worst.
As we knew that one of asthmatic trigger is allergen and it alters the releasing histamine from mast cell. Theobromine is a chemical constituent which is the same class with theophylline, xanthine, but didn’t show antiasthmatic action. Different with theophylline, theobromine has no alkyl group in N1 position, so it could be a high chance to alkylate this part by a bulky alkyl group.
Based on the Structure-Activity Relationship (SAR), the elongation of alkyl group in N1 position of theobromine can cause it having dilatation effect.
Structure-Activity Relationship of Alkylxanthine Derivatives
Then, I created the N1-alkyltheobromine by substituting isopropyl and sec-butyl group to the N1 position of theobromine. The synthesis was undergoing reaction between theobromine and alkyl bromide derivatives and catalyzed by NaOH in the ethanol as a solvent. The reaction took 36 hours heating and I have to stay in laboratory all night along.
reflux (reaction taking 36 hours)
The proton withdrawal from NaOH to imide group of Theobromine
SN2 Reaction Mechanism of Theobromine & sec-butyl bromide
After isolation using Preparative Thin Layer Chromatography (TLC) then I got the crystal as my product. Refer to the preliminary test (melting point, solubility and TLC), I confident that my product was done. I confirmed its molecule structure by running with UV, IR, NMR and GC-MS Spectroscopy method, and my goodness!!! It was absolutely perfect as what I expected.
Thin Layer Chromatography, the highest spots are my products (less polar)
UV Spectrum, any absorption in 200-40o nm showed the chromophore of my structures
IR Spectrum, showed the functional groups suitable with my structures
NMR Spectrum, showed the C-H (Hydrocarbon) environment of my structures
Mass Spectrum, confirmed my structure as N1-sec-butyl-theobromine with molecule weight = 236
After the product being confirmed, I tested its tracheospasmolytic activity against histamine in a New Guine Pig (Cavia porcella). First, I killed the animal by hitting its back head to the porcelain sink. I was shaking and nervous to do that (my first time to kill the cute animals, OMG, forgive me), especially the bleeding one. Next, I surged the neck and evacuated the trachea.
I was a ‘murder’, he3x….
The most difficult step was a trachea preparation. I had to sew the small organ without damage the smooth muscle inside.
the trachea was so small, how could I sew it??
Thereafter, the trachea was ready to set up in the instrument. The instrument name is LEVER TRANSDUCER which contains a bath which is filled by KREBS SOLUTION, that is installed to the detector and recorder. The trachea was set into the bath and added by histamine. Since histamine is an allergen, it might cause the trachea constriction which was shown by the KIMOGRAPH (the curve shows the increasing of trachea respon when the concentration of histamine being increased). Therefore, the adding of the product will make the trachea relax back and the curve will decrease gradually, proportional with the increasing of product concentration.
Kimograph, look the adding of my product decrease the curve line gradualy
The comparison of tracheospasmolytic activity between my products (N1-isopropyltheobromine & N1-sec-butyltheobromine) and theophylline. N1-sec-butyltheobromine> theophylline > N1-isopropyltheobromine
The result of this test showed that both of my product able to inhibit histamine and cause the trachea relaxing back better than theophylline. It’s the challenge to develop my product become a new antihistamine drug.
DADDY, I DEDICATE MY PRODUCTS JUST FOR YOU, MISS U ALWAYS AND WISH U REST IN PEACE…….